EA041468B1 - PSMA-TARGETED RADIOPHARMACEUTICALS FOR THE DIAGNOSIS AND TREATMENT OF PROSTATE CANCER - Google Patents

Description

State of the art

1. Field of technology.

This invention relates to PSMA-targeted radiopharmaceuticals for the diagnosis and treatment of prostate cancer.

2. Description of the prior art.

Prostate cancer is the most common cancer in men in the world and is the second leading cause of death. Prostate cancer usually develops in men over the age of 50, and the number of patients increases rapidly with age. It usually progresses slowly, but once it develops into malignant metastases, it is extremely difficult to treat. Metastases usually begin in the lymph nodes, pelvic bones, vertebrae, and bladder around prostate cancer and gradually spread throughout the body.

The prostate specific antigen test (PAP test) and digital rectal examination are currently used primarily to diagnose prostate cancer, but transrectal ultrasound, CT, MRI, and SSVT (whole body skeletal scintigraphy) are also used. Biopsies are also done to diagnose prostate cancer. However, in most cases, the accuracy of diagnosis is low, and early diagnosis of the disease is difficult. In addition, metastases are difficult to identify and difficult to distinguish from benign diseases such as prostatic hyperplasia and prostatitis.

PET (positron emission tomography) is a medical imaging technique that diagnoses a disease using a radioactive isotope with a short half-life that emits positrons. This method can be used for early diagnosis of the disease, evaluation of treatment and confirmation of metastasis/recurrence.

[ ¹⁸ F]FDG is a typical PET radiopharmaceutical used for cancer diagnosis because it can observe enhanced glucose metabolism in cancer cells. However, prostate cancer is characterized by the fact that it is difficult to detect at an early stage or diagnose the progression of the disease, because the uptake of [ ¹⁸ F]FDG is small. Choline is the material used for the biosynthesis of phosphatidylcholine, which is essential for the formation of cell membranes, and [ ¹¹ C]choline and [ ¹⁸ P]Fluorocholine are known to be more suitable for diagnosing prostate cancer than [ ¹⁸ F]FDG. However, they have low sensitivity for early detection of prostate cancer, lymph node metastases and recurrence, and poor discrimination of prostate cancer from other types of cancer.

Prostate specific membrane antigen (PSMA) is a protein that is specifically overexpressed in prostate cancer and has an enzymatic activity that degrades N-acetyl-L-aspartylglutamate (NAAL). It is known that a compound having the structure glutamic acid-urea-lysine (GML) does not degrade into NAAL analogs and binds to PSMA very selectively. To date, several compounds with GML as a basic structure have been developed, and among them, compounds labeled with F-18 (half-life: 110 min) are being developed as PET radiopharmaceuticals for diagnosing prostate cancer.

In addition to F-18, Ga-68 is a radioactive metal that emits positrons and has the property of easily complexing with a precursor-related chelator, and ^{the 68} Ga-labeled GML compound can also be used as a PET radiopharmaceutical for diagnosing prostate cancer.

In the case of ^{the 68} Ga-labeled GML compound, it can be used as a targeted therapy for prostate cancer, replacing Ga-68, a positron-emitting isotope, a therapeutically radioactive metal that emits beta rays or alpha particles. In the case of ⁶⁸ Ga-PSMA-617, ^{a 68} Ga-labeled compound, a clinical trial is ongoing in which ¹⁷⁷ Lu-PSMA-617 labeled with Lu-177 (Lutetium-177), which emits beta rays, is synthesized instead of Ga-68 and used in patients with prostate cancer. It has been reported that most of the prostate cancer that has spread throughout the body is eliminated by repeated administration 3 times.

In addition, PSMA-targeted therapy labeled with an isotope that emits alpha particles has also been developed, and since it emits more energy than beta rays, their therapeutic effect is better. Typical nuclides include Ac-225 (actinium-225), Bi-213 (bismuth-213), At-211 (astatine-211), etc. Currently, Xofigo® is used to treat prostate cancer that has metastasized to the bones, but it is an injection of ²²³ Ra-RaCl ₂ (radium dichloride), which has no therapeutic effect on prostate cancer formed outside the bones.

The present inventors have completed the present invention after confirming that the new structured PSMA target compounds labeled with radioactive metal have high binding strength and selectivity for PSMA, as well as excellent pharmacokinetic properties.

The essence of the invention

An object of the present invention is to provide a compound in which a radioactive metal conjugated chelator is conjugated with a glutamic acid-urea-lysine compound having excellent binding strength to the PSMA protein and showing excellent pharmacokinetic properties in vivo, its stereoisomer or its pharmaceutically acceptable salt.

- 1 041468

Another object of the present invention is to provide a compound in which the chelator is conjugated with a glutamic acid-urea-lysine compound having excellent binding strength to the PSMA protein and showing excellent pharmacokinetic properties in vivo, its stereoisomer or its pharmaceutically acceptable salt.

Another object of the present invention is to provide a composition for the diagnosis of prostate cancer containing the compound as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of prostate cancer, containing the compound as an active ingredient.

Another object of the present invention is the use of a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the diagnosis of prostate cancer.

Another aspect of the present invention is the use of a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of prostate cancer.

To obtain the above objects in one aspect of the present invention provides a compound represented by formula 1 below, its stereoisomer or its pharmaceutically acceptable salt.

Formula 1

In formula 1

L ₁ is -(CH2) _a -, where a is an integer 1~8;

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

R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~6;

X is a bond or -C(O)-;

W is a bond;

L2 is a bond or -(CH ₂ )d-, where d is an integer 1~8;

Tz is a link, ^NN or ;

L3 is C _1-12 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L4 is -(CH ₂ )e-, where e is an integer 1~6;

n is an integer 0~1;

R2 is hydrogen, C _1-5 straight or branched alkyl or halogen;

Y is oxygen;

Z is a chelator including a radioactive metal, where the radioactive metal is Ga-68, Cu-64 or Lu-177 and the chelator is

HO ₂ C—\ / \ CO _{2 H} ^N _N

NN _s but ₂ s— ⁷ \__/

In another aspect of the present invention, there is provided a compound represented by formula 2 below, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

Formula 2

In formula 2

L ₁ is -(CH ₂ ) _a -, where a is an integer 1~8;

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

R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~6;

X is a bond or -C(O)-;

- 2 041468

W is a bond;

L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~8;

A-HuA

Tz is a bond, N=N or ^NN ;

L ₃ is C _1-12 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L4 is -(CH ₂ )e-, where e is an integer 1~6;

n is an integer 0~1;

R2 is hydrogen, C _1-5 straight or branched alkyl or halogen;

Y is oxygen;

Z' is a chelator and a chelator is

BUT ₂ C - \ / \ / - CO ₂ H y

A , but ₂ s—/ \_/ X

In another aspect of the present invention, there is provided a prostate cancer diagnostic composition comprising an effective amount of a compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable diluent or excipient.

In another aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of prostate cancer, comprising an effective amount of a compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable diluent or excipient.

In another aspect of the present invention, the use of a compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the diagnosis of prostate cancer is provided.

In another aspect of the present invention, the use of a compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of prostate cancer is provided.

Useful effect

The compounds provided in one aspect of the present invention incorporate a carboxylic acid bound to lysine from glutamic acid-urea-lysine (GML) to form a strong salt bridge interaction with the arginine patch at the binding site of the PSMA protein, resulting in high binding capacity. These compounds are characterized by a rapid background radiation removal effect and low in vivo non-specific binding due to the hydrophilic characteristics of the carboxylic acid. In addition, the compounds are taken up at high concentrations in tumors or cancers expressing the PSMA protein by maintaining a long blood retention time.

Brief description of the drawings

In FIG. 1 is a graph showing the results of quantitative microPET/CT obtained 270 min after injection of [ ⁶⁸ Ga]1e.

In FIG. 2 is a graph showing the results of quantitative microPET/CT obtained 270 min after [ ⁶⁸ Ga]1g injection.

In FIG. 3 is a graph showing the results of quantitative microPET/CT obtained 390 min after [ ⁶⁸ Ga]1h injection.

In FIG. 4 is a graph showing the results of quantitative microPET/CT obtained 390 min after [ ⁶⁸ Ga]1k injection.

image analysis image analysis image analysis image analysis

Description of preferred options

Hereinafter, the present invention is described in detail.

The embodiments of the present invention may be modified into various other forms, and the scope of the present invention is not limited to the embodiments described below. Those skilled in the art, having an average knowledge in this area, it is well understood that the options in accordance with the present invention are given to more accurately explain the present invention. In addition, the inclusion of an element in the description does not exclude other elements, but may include other elements unless specifically stated otherwise.

In one aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of cancer, comprising a compound represented by Formula 1 below, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as an active ingredient.

- 3 041468

Formula 1

In formula 1

L ₁ is -(CH2) _a -, where a is an integer 1~8;

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

R1 is -L5-CO2H, where L5 is -(CH2) _b -, where b is an integer 1~6;

X is a bond or -C(O)-;

W is a bond;

L2 is a bond or -(CH2)d-, where d is an integer 1~8;

Aa>A

Tz is a bond, N=N or ;

L3 is C _1-12 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L ₄ is -(CH ₂ )e-, where e is an integer 1~6;

n is an integer 0~1;

R2 is hydrogen, C1.5 straight or branched alkyl or halogen;

Y is oxygen;

Z is a chelator including a radioactive metal, where the radioactive metal is Ga-68, Cu-64 or Lu-177 and the chelator is

HO ₂ C^ / \ CO ₂ H g _N NN' _{s H} q ₂ C—/ \__/

In another aspect of the present invention

L1 is -(CH ₂ ) _a -, where a is an integer 1~6;

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

R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~4;

X is a bond or -C(O)-;

W is a bond;

L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~6;

Αν^ΑΑ Α^νΑ

Tz is a bond, N=N or >

L3 is C _1-10 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L ₄ is -(CH ₂ ) _e -, where e is an integer 2~4;

n is an integer 0~1;

R2 is hydrogen, C1.3 straight or branched alkyl or halogen;

Y is oxygen;

Z is a radioactive metal chelator, where the radioactive metal is Ga-68, Cu-64 and Lu-177, and the chelator is

In another aspect of the present invention,

L1 is -(CH ₂ ) _a -, where a is an integer 2~4;

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

R1 is hydrogen or -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~2;

X is a bond or -C(O)-;

W is a bond or -NA1- where A1 is hydrogen or pyridyl;

L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~2;

Tz is the bond, NN or ^NN )

L3 is a C _1-12 straight or branched alkylene, where one or more carbon atoms in al

- 4 041468 kylene can be replaced by oxygen atoms;

L4 is -(CH2)3-;

n is an integer 0~1;

R2 is hydrogen, methyl or halo;

Y is oxygen;

Z is a radioactive metal chelator, where the radioactive metal is Ga-68, Cu-64 or Lu-177, and the chelator may be

HO ₂ C—, / \ CO ₂ H

A A

NO ₂ C~ ^/ \_/ V

In another aspect of the present invention, the compound represented by formula 1 may be any compound selected from the group consisting of the following compounds:

(3)

- 5 041468

- 6 041468

where in the above formulas, M is a radioactive metal, and the radioactive metal is as defined in formula 1.

In another aspect of the present invention, there is provided a compound represented by formula 2 below, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

- 7 041468

Formula 2

In formula 2

L1 is -(CH ₂ ) _a -, where a is an integer 1~8;

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

R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~6;

X is a bond or -C(O)-;

W is a bond;

L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~8;

or

Tz is a bond,

L3 is C _1-12 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L ₄ is -(CH ₂ )e-, where e is an integer 1~6;

n is an integer 0~1;

R2 is hydrogen, C _1-5 straight or branched alkyl or halogen;

Y is oxygen;

Z' is a chelator and a chelator is

In another aspect of the present invention,

L1 is -(CH ₂ ) _a -, where a is an integer 1~6;

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

R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~4;

X is a bond or -C(O)-;

W is a bond;

L2 is a bond or -(CH ₂ )d-, where d is an integer 1~6;

or

Tz is

L3 is C _1-10 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L4 is -(CH ₂ ) _e -, where e is an integer 2~4;

n is an integer 0~1;

R2 is hydrogen, C1.3 straight or branched alkyl or halogen;

Y is oxygen;

Z' is a chelator and a chelator is

HO ₂ C ~ \ / \ / -CO ₂ H

In another aspect of the present invention

L1 is -(CH ₂ ) _a -, where a is an integer 2~4;

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

R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~2;

X is a bond or -C(O)-;

W is a bond;

L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~2;

or

Tz is a bond,

L3 is C1. ₈ straight alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms;

L ₄ is -(CH ₂ ) ₃ -;

- 8 041468 n is an integer 0~1;

R2 is hydrogen, methyl or halo;

Y is oxygen;

Z' is a chelator and a chelator is

In another aspect of the present invention, the compound represented by formula 2 may be any compound selected from the group consisting of the following compounds:

(3)

BUT ₂ C - _x / \ / - with ₂ n

- 9 041468

- 10 041468

The compounds represented by formula 1 or formula 2 according to this invention may be used in the form of a pharmaceutically acceptable salt, in which the salt is preferably an acid addition salt formed from pharmaceutically acceptable free acids. The acid addition salt given here can be derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, and phosphorous acid; non-toxic organic acids such as aliphatic mono/dicarboxylate, phenyl-substituted alkanoate, hydroxyalkanoate, alkanedioate, aromatic acids and aliphatic/aromatic sulfonic acids; or organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid and fumaric acid. Pharmaceutically non-toxic salts are sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrophosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutylate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, kabakat, fumarate, maliat, butyne 1,4-dioat, hexane-1,6-dioat, benzoate, chlorobenzoate, methyl benzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutylate, citrate, lactate, hydroxybutylate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate and almonds.

The acid addition salt in the present invention can be prepared in a conventional manner known to those skilled in the art. For example, the derivative represented by formula 1 or formula 2 is dissolved in an organic solvent such as methanol, ethanol, acetone, methylene chloride and acetonitrile, to which an organic acid or inorganic acid is added to cause precipitation. The precipitate is then filtered and dried to give the salt. Or, the solvent and excess acid are distilled off under reduced pressure and dried to give a salt. Or the precipitate is crystallized in an organic solvent to obtain the same.

A pharmaceutically acceptable metal salt can be prepared using a base. An alkali metal or alkaline earth metal salt is prepared by the following methods: by dissolving the compound in an excess solution of an alkali metal or alkaline earth metal hydroxide; filtering the salt of the insoluble compound; by evaporating the remaining solution and drying it. At this time, the metal salt is preferably prepared in a pharmaceutically acceptable sodium, potassium or calcium salt form. The corresponding silver salt is prepared by reacting an alkali or alkaline earth metal salt with the corresponding silver salt (eg silver nitrate).

In addition, the present invention includes not only a compound represented by formula 1 or formula 2, but also a pharmaceutically acceptable salt thereof, an optical isomer, optionally derived therefrom.

In another aspect of the present invention, the present invention provides a prostate cancer diagnostic composition comprising an effective amount of a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as an active ingredient, and

- 11 041468 pharmaceutically acceptable diluent or excipient.

The composition for diagnosing prostate cancer can diagnose prostate cancer by selectively binding the compound to PSMA (Prostate Specific Membrane Antigen) overexpressed in prostate cancer cells.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of prostate cancer, comprising a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable diluent or excipient.

The compound represented by formula 1 and a pharmaceutically acceptable salt thereof can be administered orally or parenterally and used in general pharmaceutical composition forms. Those. the compound represented by formula 1 and its pharmaceutically acceptable salt can be prepared for oral or parenteral administration by mixing with commonly used diluents or excipients such as fillers, diluents, binding agents, wetting agents, disintegrants and surfactants. Solid oral formulations are tablets, pills, powders, granules and capsules. These solid formulations are prepared by mixing one or more compounds according to the invention with one or more suitable excipients such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. With the exception of simple excipients, lubricating agents such as magnesium stearate, talc, etc. may be used. Liquid formulations for oral administration are suspensions, solutions, emulsions and syrups, and the aforementioned formulations may contain various excipients such as wetting agents, sweeteners, flavors and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Compositions for parenteral administration are sterilized aqueous solutions, water-insoluble excipients, suspensions and emulsions. Water-insoluble excipients and suspensions may contain, in addition to the active compound or compounds, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethylolate, etc.

Pharmaceutical compositions containing a compound represented by formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient according to the present invention may be administered parenterally, and parenteral administration includes subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection.

To prepare the composition as a formulation for parenteral administration, the compound represented by formula 1, or a pharmaceutically acceptable salt thereof according to the present invention, is mixed with a stabilizer or buffer agent to obtain a solution or suspension, which is then formulated into ampoules or vials. The composition of the present invention may be sterilized and further comprises preservatives, stabilizers, wettable powders or emulsifiers, salts and/or buffers for adjusting osmotic pressure and other therapeutically useful materials, and the composition may be formulated by conventional mixing, granulation or coating.

Formulations for oral administration include tablets, pills, hard/soft capsules, solutions, suspensions, emulsions, syrups, granules, elixirs and lozenges, and the like. These formulations may include diluents (eg lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine) and lubricating agents (eg silica, talc, stearate and its magnesium or calcium salt and/or polyethylene glycol) in addition to active ingredient.

Tablets may include binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone and, if necessary, disintegrants such as starch, agarose, alginic acid or its sodium salt or azeotropic mixtures, and/or they may additionally include absorbents, dyes, flavors and sweeteners.

Hereinafter, the present invention will be described in detail using the following examples.

However, the following examples are only intended to illustrate the present invention and the content of the present invention is not limited to them.

Example 1 Preparation of compounds 3b and 3c.

Preparation of compound 3b.

Compound 3a (5.2 g, 10.66 mmol) was dissolved in dichloromethane (100 ml) and cooled to 0°C, there

- 12 041468 slowly add tert-butyl bromoacetate (1.9 ml, 12.8 mmol). The mixture was kept at 0°C, triethylamine (2.2 ml, 16 mmol) was slowly added thereto, and the mixture was stirred while gradually raising the temperature to room temperature. After the mixture was stirred for 3 hours, water (50 ml) was added thereto and the organic compound was extracted with dichloromethane (50 ml, twice). The collected organic layer was treated with anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (5% methanol/dichloromethane) to give compound 3b (3.36 g, 52%).

1H NMR (400 MHz, CDCl ₃ ) δ 1.39-1.53 (m, 36H), 1.55-1.89 (m, 5H), 2.02-2.10 (m, 1H), 2 .22-2.37 (m, 2H), 2.54-2.58 (m, 2H), 3.27 (s, 2H), 4.28-4.36 (m, 2H), 5.07 -5.10 (m, 2H);

¹³ C NMR (100 MHz, CDCl ₃ ) δ 22.6, 27.9, 28.0, 28.1, 28.2, 28.5, 29.6, 31.6, 32.8, 49.0 , 51.7, 53.0, 53.5, 80.5, 81.1, 81.6, 82.0, 156.8, 171.9, 172.1, 172.4, 172.5;

MS (ESI) m/z 602 [M+H]+.

Preparation of Compound 3 c.

Compound 3a (500 mg, 1.03 mmol) was dissolved in ethanol (10 ml), then stirred at 0° C. for 10 min. tert-Butyl acrylate (0.38 ml, 2.58 mmol) was slowly added thereto, followed by stirring at 0° C. for 20 h. After completion of the reaction, the solvent was removed and the concentrate was separated by column chromatography (8% methanol/dichloromethane) to give compound 3c (0.23 g, 37%).

1H NMR (400 MHz, CDCI3) δ 1.40 (s, 9H), 1.41 (s, 9H), 1.43 (s, 18H), 1.48-1.65 (m, 3H), 1 .70-1.86 (m, 2H), 2.00-2.07 (m, 1H), 2.21-2.36 (m, 2H), 2.48 (t, J=6.6 Hz , 2H), 2.58-2.69 (m, 2H), 2.86 (t, J=6.6 Hz, 2H), 4.26-4.34 (m, 2H), 5.26 ( dd, J=13.0, 8.2 Hz, 2H); MS (ESI) m/z 616 [M+H]+

Example 2 Preparation of Compounds 2a and 2b.

Stage 1. Preparation of compound 5a.

Triphosgene (107 mg, 0.36 mmol) was dissolved in acetonitrile (5.0 ml), compound 3a (500 mg, 1.03 mmol) dissolved in acetonitrile at 0°C was slowly added thereto. Triethylamine (0.50 ml, 3.61 mmol) was then added thereto, followed by stirring for 30 minutes. Propagylamine (4a, 0.072 ml, 1.13 mmol) was added thereto at 0°C. After 15 minutes, the mixture was stirred at room temperature for 1 hour, concentrated under reduced pressure, and then water was added thereto. The organic compound is re-extracted 3 times using ethyl acetate. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (2% methanol/dichloromethane) to give compound 5a (492 mg, 84%) as a white solid.

1H NMR (400 MHz, CDCl ₃ ) δ 1.25-1.30 (m, 2H), 1.44 (s, 18H), 1.48 (s, 9H), 1.51-1.60 (m , 3H), 1.67-1.76 (m, 1H), 1.80-1.90 (m, 1H), 2.05-2.13 (m, 1H), 2.18 (t, J =2.6 Hz, 1H), 2.29-2.40 (m, 2H), 3.06-3.12 (m, 1H), 3.30-3.36 (m, 1H), 3, 95-4.06 (m, 2H), 4.08-4.14 (m, 1H), 4.36 (sext, J=4.4 Hz, 1H), 5.64 (d, J=7, 6 Hz, 1H), 5.69 (t, J=5.2 Hz, 1H), 5.89 (t, J=5.4 Hz, 1H), 6.11 (d, J=8.4 Hz , 1H);

¹³ C NMR (100 MHz, CDCl ₃ ) δ 23.4, 27.7, 27.8, 27.9, 28.0, 29.6, 29.7, 31.7, 32.1, 39.4 , 53.3, 54.2, 70.5, 80.7, 81.4, 81.5, 83.1, 158.0, 158.2, 172.0, 172.3, 174.6;

MS (ESI) m/z 569 [M+H] ⁺ .

Step 1: Preparation of Compound 5b.

Compound 4b (200 mg, 1.51 mmol) was dissolved in acetonitrile (5.0 ml), 4-nitrophenyl chloroformate (305 mg, 1.51 mmol) was slowly added thereto at 0°C. Triethylamine (0.50 ml, 3.61 mmol) was added thereto, followed by stirring for 30 minutes. Compound 3a (886 mg, 1.82 mmol) dissolved in acetonitrile (10 ml) was slowly added thereto at 0° C., diisopropylethylamine (0.324 ml, 1.82 mmol) was added thereto. After 15 minutes, the mixture was stirred at 100° C. for 12 hours. After the mixture was cooled to room temperature, water was added thereto. The organic compound is re-extracted 3 times using ethyl acetate. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (5% methanol/dichloromethane) to give compound 5b (836 mg, 86%) as a colorless liquid.

1H NMR (400 MHz, CDCl ₃ ) δ 1.27-1.37 (m, 2H), 1.43 (s, 9H), 1.45 (s, 18H), 1.50-1.55 (m , 2H), 1.59-1.65

- 13 041468 (m, 1H), 1.72-1.88 (m, 2H), 2.01-2.10 (m, 1H), 2.27-2.34 (m, 1H), 2, 35(t, J=2.4Hz, 1H), 2.16(kv, J=6.7Hz, 2H), 4.25-4.34(m, 2H), 4.50(ddd, J =25.2, 18.0, 2.4Hz, 2H), 5.21(t, J=5.8Hz, 1H), 5.48(s, 1H), 5.50(s, 1H) , 7.32 (dd, J=4.8, 1.6 Hz, 2H), 8.59 (d, J=6.4 Hz, 2H);

¹³ C NMR (100 MHz, CDQ3) δ 22.4, 27.9, 28.0, 28.1, 28.3, 29.4, 31.6, 32.4, 38.2, 40.7, 52.9, 53.3, 72.9, 79.3, 80.5, 81.6, 82.0, 119.5, 149.6, 151.2, 155.3, 157.1, 172, 3, 172.4, 172.5;

MS (ESI) m/z 646 [M+H]+.

Step 2: Preparation of Compound 6a.

Compound 5a (0.8 g, 1.4 mmol) and 2-aminoethyl, 2'-azidoethyl ether (0.37 g, 2.81 mmol) are dissolved in ethanol (20 ml), 1 M CuSO ₄ (0 .28 ml, 0.28 mmol) and 2 M sodium ascorbate (0.21 ml, 0.42 mmol), then stirred for 1 hour. The reagent was filtered and the solvent was removed under reduced pressure. The concentrate was separated by NH silica gel column chromatography (2% methanol/dichloromethane) to give compound 6a (0.45 g, 46%).

MS (ESI) m/z 699 [M+H] ⁺ .

Stage 2. Preparation of compound 6.

Compound 6b (450 mg, 42%) is prepared according to the procedure described for compound 6a, except that compound 5b (880 mg, 1.4 mmol), 2-aminoethyl, 2'-azidoethyl ether (0.26 g, 2.00 mmol), 1 M CuSO ₄ (0.27 ml, 0.27 mmol) and 2 M sodium ascorbate (0.20 ml, 0.41 mmol).

MS (ESI) m/z 776 [M+H] ⁺ .

Step 3: Preparation of Compound 7a.

DOTA-tris(tBu) ether (0.44 g, 0.77 mmol) was dissolved in dichloromethane (15 ml), hydroxybenzotriazole (HOBt, 0.13 g, 0.97 mmol), TBTU (0.31 g, 0.97 mmol) and diisopropylethylamine (0.224 ml, 0.13 mmol), then stirred at room temperature for 10 minutes. Compound 6a (0.45 g, 0.64 mmol) dissolved in dichloromethane (5 ml) was added thereto, followed by stirring for 1 h. The reaction was stopped by adding water (20 ml) and the organic compound was extracted using dichloromethane (20 ml x 2) . The organic solvent was concentrated under reduced pressure, and the concentrate was separated by column chromatography (4% methanol/dichloromethane) to give compound 7a (0.32 g, 40%).

1H NMR (400 MHz, methanol-d ₄ ) δ 1.45-1.50 (m, 69H), 1.52-1.74 (m, 4H), 1.74-1.85 (m, 2H) , 1.98-2.09 (m, 4H), 2.25-2.38 (m, 6H), 3.09-3.16 (m, 6H), 3.35-3.43 (m, 4H), 3.47-3.56 (m, 4H), 3.61-3.68 (m, 2H), 3.81-3.86 (m, 4H), 4.11-4.15 ( m, 2H), 4.18-4.25 (m, 2H), 4.36-4.38 (m, 4H), 4.55 (t, J=4.8 Hz, 4H), 7.84 (s, 0.7H), 7.86 (s, 0.3H);

MS (ESI) m/z 1254 [M+H] ⁺ .

Step 3: Preparation of Compound 7b.

Compound 7b (0.15 g, 29%) was prepared as described for compound 7a, except that DOTA-tris(tBu) ester (270 mg, 0.46 mmol), hydroxybenzotriazole (HOBT, 0.078 g , 0.58 mmol), TBTU (0.19 g, 0.58 mmol), diisopropylethylamine (0.134 ml, 0.77 mmol) and compound 6b (0.30 g, 0.39 mmol).

1H NMR (400 MHz, methanol-d ₄ ) δ 1.35-1.49 (m, 63H), 1.60-1.68 (m, 1H), 1.73-1.85 (m, 4H) , 1.99-2.08 (m, 3H), 2.27-2.34 (m, 4H), 3.20-3.26 (m, 8H), 3.51 (t, J=5, 6 Hz, 4H), 3.69-3.78 (m, 5H), 3.81-3.83 (m, 1H), 4.09-4.23 (m, 1H), 4.46-4 .56 (m, 4H), 5.03 (s, 4H), 7.41 (d, J=6.8 Hz, 2H), 7.94 (s, 1H), 8.43 (d, J= 6.4 Hz, 2H);

MS (ESI) m/z 1331 [M+H] ⁺ .

Step 4: Preparation of Compound 2a.

Compound 7a (300 mg, 0.24 mmol) was added to 70% trifluoroacetic acid/dichloromethane (6 ml), then stirred for 5 hours. Diethyl ether (20 ml) was added thereto to obtain a precipitate, and it was separated using a centrifuge. The mixture was separated by HPLC and dried using a lyophilizer to give Compound 2a (115 mg, 52%) as a solid.

1H NMR (400 MHz, D2O) δ 1.31-1.44 (m, 2H), 1.45-1.55 (m, 2H), 1.66-1.75 (m, 1H), 1, 79-1.88 (m, 1H), 1.93-2.02 (m, 1H), 2.14-2.22 (m, 1H), 2.52 (t, J=7.2 Hz, 2H), 3.11 (t, J=6.8 Hz, 3H), 3.14-3.55 (m, 26H), 3.58 (t, J=5.2 Hz, 3H), 3, 62-3.93 (m, 6H), 3.96 (t, J=5.6 Hz, 4H), 4.18 (dd, J=13.6, 4.8 Hz, 1H), 4.27 (dd, J=14.4, 5.2 Hz, 1H), 4.39 (s, 2H), 4.61 (t, J=5.2 Hz, 2H), 7.93 (s, 1H) ;

MS (ESI) m/z 918 [M+H] ⁺ .

Step 4: Preparation of Compound 2b.

Compound 2b (10 mg, 48%) was prepared as described for compound 2a as a solid, except that compound 7a (28 mg, 21 µmol) and 70% trifluoroacetic acid/dichloromethane (0.4 ml ).

1H NMR (400 MHz, D ₂ O) δ 1.31-1.44 (m, 4H), 1.51-1.62 (m, 2H), 1.64-1.75 (m, 1H), 1.79-1.87 (m, 1H), 1.90-1.99 (m, 1H), 2.11-2.19 (m, 1H), 2.49 (t, J=7.6 Hz, 2H), 2.90-3.48 (m, 18H), 5.52 (t, J=5.2 Hz, 3H), 3.61-3.88 (m, 6H), 3.92 (t, J=4.8 Hz, 3H), 4.16 (dd, J=14.0, 5.2 Hz, 1H), 4.25 (dd, J=14.4, 5.2 Hz, 1H), 4.59 (t, J=4.4 Hz, 2H), 5.19 (s, 2H), 7.60 (d, J=7.2 Hz, 2H), 8.03 (s, 1H), 8.42 (d, J=7.2 Hz, 2H);

MS (ESI) m/z 995 [M+H] ⁺ .

- 14 041468

Example 3 Preparation of compounds 2c and 2d.

Step 1: Preparation of compound 5c.

4-Pentaenoic acid (82 mg, 0.83 mmol) was dissolved in dichloromethane (10 ml) and cooled to 0°C, N,N'-dicyclohexylcarbodiimide (190 mg, 0.91 mmol) and compound 3c (0. 5 g, 0.83 mmol), then stirred at room temperature for 1 hour. The organic layer was filtered several times and the solvent was removed under reduced pressure. The concentrate was separated by column chromatography (30% ethyl acetate/n-hexane) to give compound 5c (0.29 g, 52%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.34-1.67 (m, 39H), 1.68-2.02 (m, 5H), 2.16-2.32 (m, 2H), 2.37-2.56 (m, 5H), 3.22 (t, J=7.2 Hz, 1H), 3.29 (t, J=7.6 Hz, 1H), 3.82-3 .90 (m, 2H), 4.17-4.29 (m, 2H), 5.49-5.52 (m, 1.5H), 5.60 (d, J=8.0 Hz, 0 .5 Hz);

MS (ESI) m/z 704 [M+Na]+.

Step 1. Preparation of compound 5d.

Compound 5d (0.18 g, 79%) was prepared as described for compound 5c, except that 4-pentaenoic acid (32 mg, 0.32 mmol), N,N'-dicyclohexulcarbodiimide (74 mg , 0.36 mmol) and compound 3c (0.20 g, 0.32 mmol).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.41-1.43 (m, 27), 1.44 (s, 9H), 1.51-1.63 (m, 3H), 1.76- 1.88 (m, 2H), 1.93-1.96 (m, 1H), 2.01-2.08 (m, 1H), 2.20-2.36 (m, 2H), 2, 46-2.53(m, 5H), 2.57-2.60(m, 1H), 3.26(dt, J=21.2, 7.7Hz, 2H), 3.52(q, J=7.2Hz, 2H), 4.24-4.35(m, 2H), 5.05(dd, J=16.4, 8.0Hz, 1H), 5.33(dd, J =61.2, 8.0 Hz, 1H);

MS (ESI) m/z 718 [M+Na]+.

Step 2: Preparation of compound 6c.

Compound 5c (0.26 g, 0.38 mmol) and 2-aminoethyl, 2'-azidoethyl ether (60 mg, 0.46 mmol) are dissolved in ethanol (5 ml), 1 M CuSO ₄ (0.076 ml, 0.076 mmol) and 2 M sodium ascorbate (0.057 ml, 0.11 mmol), then stirred for 1 hour. The reagent was filtered and the solvent was removed under reduced pressure. The concentrate was separated by NH silica gel column chromatography (3% methanol/dichloromethane) to give compound 6c (0.27 g, 87%).

1H NMR (400 MHz, methanol^) δ 1.37-1.55 (m, 36H), 1.56-1.70 (m, 2H), 1.71-1.94 (m, 2H), 1 .95-2.14 (m, 2H), 2.24-2.40 (m, 2H), 2.58-2.91 (m, 2H), 2.92-3.12 (m, 2H) , 3.33-3.48 (m, 4H), 3.49-3.76 (m, 4H), 3.77-3.92 (m, 2H), 3.96 (s, 1H), 4 .45-4.28 (m, 3H), 4.46-4.65 (m, 1H);

MS (ESI) m/z 813 [M+H] ⁺ .

Step 2: Preparation of Compound 6d.

Compound 6d (60.0 mg, 50%) was prepared as described for compound 6c, except compound 5d (0.10 g, 0.14 mmol), 2-aminoethyl, 2'-azidoethyl ester was used. (21 mg, 0.16 mmol), 1 M CuSO4 (0.030 ml, 0.030 mmol) and 2 M sodium ascorbate (0.020 ml, 0.040 mmol).

1H NMR (400 MHz, CDCl ₃ ) δ 1.23-1.30 (m, 2H), 1.40 (s, 18H), 1.42 (s, 18H), 1.68 (s, 6H), 1.74-1.87 (m, 2H), 1.99-2.09 (m, 1H), 2.24-2.35 (m, 2H), 2.41-2.47 (m, 2H ), 2.70-2.75 (m, 1H), 2.96-3.08 (m, 2H), 3.20-3.31 (m, 2H), 3.28-3.54 (m , 3H), 3.81 (t, J=8.0 Hz, 2H), 4.24-4.42 (m, 2H), 4.47-4.55 (m, 2H), 5.59 ( dd, J=53.4, 7.4 Hz, 1H), 5.77 (dd, J=37.6, 8.4 Hz, 1H), 7.53 (d, J=16.4 Hz, 1H );

MS (ESI) m/z 826 (M+H) ⁺ .

Step 3 Preparation of Compound 7c.

DOTA-tpuc(IBu) ester (84 mg, 0.015 mmol) was dissolved in dichloromethane (5 ml), hydroxybenzotriazole (HOBt, 25 mg, 0.019 mmol), TBTU (59 mg, 0.019 mmol) and diisopropylethylamine (0.042 ml, 0 .25 mmol), then stirred at room temperature for 10 min. Compound 6c (100 mg, 0.12 mmol) dissolved in dichloromethane (2 ml) was added thereto, followed by stirring for 1 h. The reaction was quenched by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 ml x 2). The reagent is dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was separated by column chromatography (3% methanol/dichloromethane) to give compound 7c (95 mg, 56%).

- 15 041468 ¹ H NMR (400 MHz, methanol^) δ 1.42-1.65 (m, 63H), 1.71-1.85 (m, 2H), 1.95-3.69 (m, 40H), 3.74 (s, 3H), 3.79-3.92 (m, 2H), 3.96 (s, 1H), 4.11-4.20 (m, 3H), 4.50 -4.58 (m, 2H);

MS (ESI) m/z 1388 [M+Na]+.

Step 3: Preparation of Compound 7d.

Compound 7d (51 mg, 61%) was prepared as described for compound 7c, except that DOTA-tpuc(IBu) ether (29 mg, 0.073 mmol) dissolved in dichloromethane (5 mL) and hydroxybenzotriazole ( HOBt, 12 mg, 0.091 mmol), TBTU (29 mg, 0.091 mmol), diisopropylethylamine (15.86 μΓ, 91.07 μmol) and compound 6d (50 mg, 60.5 μmol).

1H NMR (400 MHz, CDCh) δ 0.75-0.94 (m, 2H), 1.23-1.61 (m, 63H), 1.69 (s, 5H), 1.77-1, 87(m, 2H), 2.00-2.08(m, 3H), 2.21(brs, 2H), 2.27-2.37(m, 3H), 2.41-2.48( m, 4H), 2.78 (s, 4H), 2.94-3.06 (m, 3H), 3.20-3.38 (m, 5H), 3.43-3.56 (m, 4H), 3.60 (t, J=5.2 Hz, 2H), 3.66-3.75 (m, 5H), 3.80 (t, J=4.8 Hz, 2H), 4, 19 (d, J=4.0 Hz, 2H), 4.25-4.34 (m, 2H), 4.50-4.54 (m, 2H), 5.47 (dd, J=26, 8, 8.0 Hz, 1H), 5.66 (dd, J=12.4, 8.4 Hz, 1H), 7.71 (d, J=41.2 Hz, 1H);

MS (ESI) m/z 1381 [M+H] ⁺ .

Step 4 Preparation of Compound 2c.

Compound 7c (60 mg, 0.044 mmol) was added to 70% trifluoroacetic acid/dichloromethane (2 ml), then stirred for 4 hours. Diethyl ether (20 ml) was added thereto to obtain a precipitate, and it was separated using a centrifuge. The mixture was separated by HPLC and dried using a lyophilizer to give compound 2c (25 mg, 58%) as a solid.

1H NMR (400 MHz, D2O) δ 1.10-1.30 (m, 2H), 1.31-1.50 (m, 2H), 1.52-1.63 (m, 1H), 1, 64-1.76 (m, 1H), 1.78-1.89 (m, 1H), 1.99-2.09 (m, 1H), 2.36-2.40 (m, 2H), 2.62-2.65 (m, 1H), 2.77-2.80 (m, 2H), 2.95-2.98 (m, 3H), 3.00-3.19 (m, 7H ), 3.21-3.42 (m, 11H), 3.46-3.47 (m, 3H), 3.49-3.72 (m, 4H), 3.82-3.86 (m , 3H), 3.95 (s, 2H), 4.01-4.15 (m, 4H), 4.53-4.56 (m, 2H), 7.84 (s, 1H);

MS (ESI) m/z 974 [M+H] ⁺ .

Step 4 Preparation of Compound 2d.

Compound 2d (19 mg, 66%) was prepared as described for compound 2c as a solid, except compound 7d (40 mg, 0.029 mmol) was used.

1H NMR (400 MHz, D2O) δ 1.25-1.42 (m, 2H), 1.44-1.64 (m, 2H), 1.65-1.76 (m, 1H), 1, 78-1.91 (m, 1H), 1.92-2.04 (m, 1H), 2.14-2.22 (m, 0.5H), 2.52 (t, J=7.2 Hz, 2H), 2.59 (t, J=7.2 Hz, 1.5H), 2.64 (t, J=7.2 Hz, 1H), 2.81 (t, J=7.2 Hz, 1H), 2.88 (t, J=7.2 Hz, 1H), 3.03-3.07 (m, 3H), 3.08-3.54 (m, 19H), 3.55 -3.65(m, 7H), 3.66-3.87(m, 4H), 3.96(t, J=4.8Hz, 4H), 4.17-4.22(m, 1H ), 4.25-4.28 (m, 1H), 4.62-4.64 (m, 2H), 7.92 (s, 0.6H), 7.93 (s, 0.4H);

MS (ESI) m/z 974 [M+H] ⁺ .

Example 4 Preparation of compound 2e.

Step 1: Preparation of Compound 9a.

Compound 3b (600 mg, 0.997 mmol) synthesized in Example 1 was dissolved in dichloromethane (10 ml), N,N'-dicyclohexylcarbodiimide (DCC, 226 mg, 1.04 mmol) was slowly added thereto at room temperature. 2-(2-(2-azidoethoxy)ethoxy)acetic acid 8a (N ₃ -(CH ₂ CH ₂ O) ₂ -CH ₂ COOH, 226 mg, 1.20 mmol) was slowly added thereto, then stirred for 1 h Water is added to the reagent and then the organic compound is re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (60% ethyl acetate/n-hexane) to give

- 16 041468 compound 9a (520 mg, 67%) as a colorless liquid.

1H NMR (400 MHz, methanol^) δ 1.44-1.49 (m, 36H), 1.50-1.57 (m, 2H), 1.58-1.71 (m, 2H), 1 .73-1.84 (m, 2H), 2.00-2.09 (m, 1H), 2.25-2.38 (m, 2H), 3.33-3.39 (m, 4H) , 3.65-3.72 (m, 6H), 3.96 (d, J=1.2 Hz, 1H), 4.11-4.22 (m, 3H), 4.33 (s, 2H ), 6.32-6.36 (m, 1H);

MS (ESI) m/z 773 [M+H]+.

Step 2: Preparation of Compound 10a.

Compound 9a (490 mg, 0.634 mmol) synthesized in Step 1 above was dissolved in ethanol (20 ml), 10% palladium-carbon (67 mg) was added thereto, followed by stirring for 12 hours under hydrogen. The reaction solution is filtered, washed with ethanol, and concentrated under reduced pressure. The concentrate was separated by column chromatography (4% methanol/dichloromethane, NH silica gel) to give compound 10a (425 mg, 90%) as a colorless liquid.

1H NMR (400 MHz, methanol^) δ 1.34-1.39 (m, 2H), 1.44-1.49 (m, 36H), 1.51-1.65 (m, 4H), 1 .73-1.84 (m, 2H), 2.00-2.07 (m, 1H), 2.31 (q, J=6.8 Hz, 2H), 2.80 (t, J=5 .2 Hz, 2H), 3.33-3.40 (m, 1H), 3.52 (kv, J=5.2 Hz, 2H), 3.61-3.66 (m, 3H), 3 .69-3.71 (m, 1H), 3.97 (d, J=1.2 Hz, 1H), 4.11 (s, 1H), 4.13-4.21 (m, 2H), 4.32 (s, 2H);

MS (ESI) m/z 747 [M+H] ⁺ .

Step 3. Preparation of compound 11a.

DOTA-tPuc(IBu) ester (55 mg, 0.096 mmol) was dissolved in dichloromethane (2.0 ml) to which were added hydroxybenzotriazole (HOBt, 22 mg, 0.160 mmol), TBTU (52 mg, 0.160 mmol) and diisopropylethylamine (0.042 ml , 0.241 mmol), then stirred at room temperature for 10 minutes. Compound 10a (60 mg, 0.080 mmol) synthesized in Step 2 above, dissolved in dichloromethane (2.0 ml) was added thereto, followed by stirring at room temperature for 1 hour. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (3% methanol/dichloromethane) to give compound 11a (75 mg, 71%) as a colorless liquid.

1H NMR (400 MHz, methanol^) δ 1.44-1.50 (m, 63H), 1.54-1.65 (m, 4H), 1.73-1.82 (m, 2H), 2 .00-2.09 (m, 3H), 2.15-2.34 (m, 6H), 2.59-3.25 (br, 16H), 3.38-3.40 (m, 2H) , 3.55-3.57 (m, 3H), 3.62 (s, 2H), 3.63-3.69 (m, 3H), 3.97 (s, 2H), 4.08 (s , 2H), 4.09-4.21 (m, 3H), 4.31 (s, 2H);

MS (ESI) m/z 1302 [M+H] ⁺ .

Step 4 Preparation of Compound 2e.

Compound 11a (50 mg, 0.038 mmol) synthesized in step 3 above was dissolved in 70% trifluoroacetic acid/dichloromethane (0.5 ml), then stirred at room temperature for 4 h. The reagent was concentrated under reduced pressure and separated with high efficiency liquid chromatography (HPLC) to give compound 2e (26 mg, 74%) as a white solid.

1H NMR (400 MHz, D2O) δ 1.16-1.30 (m, 2H), 1.36-1.50 (m, 2H), 1.52-1.62 (m, 1H), 1, 64-1.73(m, 1H), 1.76-1.86(m, 1H), 1.98-2.06(m, 1H), 2.35(td, J=7.2, 1 .6 Hz, 2H), 2.86-3.38 (m, 20H), 3.48-3.60 (m, 10H), 3.70-3.91 (br, 3H), 3.96 ( s, 2H), 4.00-4.12 (m, 3H), 4.25 (s, 2H);

MS (ESI) m/z 910 [M+2H]+.

Example 5 Preparation of compounds 2f, 2g and 2h.

ISc, R3 = I 2h, R.-l

Stage 1. Preparation of compound 12a.

Lysine (Fmoc-Lys(Z)-OH, 275 mg, 0.546 mmol) was dissolved in dichloromethane (10 mL) and added

- 17 041468 hydroxybenzotriazole (HOBt, 123 mg, 0.1910 mmol), TBTU (292 mg, 0.910 mmol) and diisopropylethylamine (0.238 ml, 1.37 mmol), then stirred at room temperature for 10 minutes. Compound 10a (340 mg, 0.455 mmol) synthesized in step 2 of Example 4 was added thereto, dissolved in dichloromethane (5.0 ml), then stirred at room temperature for 1 hour. Water was added to the reagent, and then the organic compound was re-extracted 3 times with dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (2% methanol/dichloromethane). Dichloromethane (15 ml) was added to the resulting compound, piperidine (0.043 ml, 0.438 mmol) was added thereto, followed by stirring at room temperature for 24 hours. The reagent was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane, NH silica gel) to give compound 12a (480 mg, 84%) as a colorless liquid.

1H NMR (400 MHz, methanol ₄ ) δ 1.34-1.39 (m, 2H), 1.44-1.48 (m, 36H), 1.50-1.70 (m, 10 H), 1.72-1.84 (m, 2H), 2.00-2.08 (m, 1H), 2.24-2.38 (m, 2H), 3.11 (t, J=6.8 Hz, 2H), 3.33-3.41 (m, 4H), 3.55 (kv, J=5.2 Hz, 2H), 3.61-3.68 (m, 4H), 3.96 (d, J=1.6 Hz, 1H), 4.08 (d, J=1.2 Hz, 1H), 4.10-4.21 (m, 3H), 4.31 (s, 1H) , 5.06 (s, 2H), 7.27-7.32 (m, 1H), 7.33-7.34 (m, 4H);

MS (ESI) m/z 1010 [M+H]+.

Stage 2. Obtaining compound 13a.

DOTA-tris(tBu) ether (211 mg, 0.369 mmol) was dissolved in dichloromethane (10 mL) to which were added hydroxybenzotriazole (HOBt, 83 mg, 0.614 mmol), TBTU (197 mg, 0.614 mmol) and diisopropylethylamine (0.161 mL, 0.921 mmol), then stirred at room temperature for 10 min. Compound 12a (310 mg, 0.307 mmol) synthesized in Step 1 above, dissolved in dichloromethane (5.0 ml) was added thereto, followed by stirring at room temperature for 1 hour. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (5% methanol/dichloromethane) to give compound 13a (323 mg, 67%) as a colorless liquid.

1H NMR (400 MHz, methanol-d ₄ ) δ 1.36-1.39 (m, 2H), 1.44-1.49 (m, 63H), 1.51-1.73 (m, 8H) , 1.75-1.84 (m, 2H), 2.00-2.07 (m, 3H), 2.08-2.26 (m, 4H), 2.28-2.36 (m, 3H), 2.38-3.05(brs, 12H), 3.11(t, J=6.6Hz, 2H), 3.16-3.28(m, 4H), 3.36(t , J=6.6Hz, 2H), 3.38-3.52(brs, 3H), 3.54(kv, J=4.0Hz, 2H), 3.58-3.68(m, 5H), 3.97 (d, J=4.4 Hz, 1H), 4.07 (s, 1H), 4.09-4.22 (m, 3H), 4.26-4.28 (m , 1H), 4.31 (s, 1H), 5.06 (s, 2H), 7.26-7.32 (m, 1H), 7.33-7.38 (m, 4H);

MS (ESI) m/z 1565 [M+2H]+.

Stage 3. Preparation of compound 14a.

Compound 13a (300 mg, 0.192 mmol) synthesized in Step 2 above was dissolved in ethanol (20 ml), 10% palladium-carbon (20 mg) was added thereto, followed by stirring for 2 hours under hydrogen. The reaction solution is filtered, washed with ethanol, and concentrated under reduced pressure. The concentrate was separated by column chromatography (4% methanol/dichloromethane, NH silica gel) to give compound 14a (260 mg, 95%) as a colorless liquid.

1H NMR (400 MHz, methanol-d ₄ ) δ 1.33-1.42 (m, 4H), 1.44-1.49 (m, 63H), 1.51-1.57 (m, 4H) , 1.59-1.73(m, 4H) 1.74-1.85(m, 3H), 2.00-2.08(m, 3H), 2.09-2.27(brs, 4H ), 2.29-2.38 (m, 3H), 2.60-2.65 (m, 2H), 2.68 (t, J=7.2 Hz, 2H), 2.73-2, 94 (brs, 7H), 3.05-3.17 (brs, 3H), 3.25-3.28 (m, 2H), 3.34-3.39 (m, 2H), 3.43 ( brs, 1H), 3.47-3.39 (m, 1H), 3.53-3.57 (m, 3H), 3.63 (s, 2H), 3.65-3.68 (m, 2H), 3.98 (d, J=5.6 Hz, 1H), 4.09 (s, 1H), 4.11-4.22 (m, 3H), 4.32 (s, 2H);

MS (ESI) m/z 1430 [M+H] ⁺ .

Step 4: Preparation of Compound 15a.

4-Phenylbutyric acid (8.4 mg, 0.050 mmol) was dissolved in dichloromethane (1.0 ml) to which were added hydroxybenzotriazole (HOBt, 11 mg, 0.084 mmol), TBTU (27 mg, 0.084 mmol) and diisopropylethylamine (0.022 ml, 0.126 mmol), then stirred at room temperature for 10 minutes. Compound 14a (60 mg, 0.042 mmol) synthesized in Step 3 above, dissolved in dichloromethane (1.0 ml) was added thereto, followed by stirring at room temperature for 2 hours. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (4% methanol/dichloromethane) to give compound 15a (17 mg, 26%) as a colorless liquid.

1H NMR (400 MHz, methanol-d ₄ ) δ 1.44-1.48 (m, 63H), 1.51-1.71 (m, 6H), 1.74-1.84 (m, 4H) , 1.86-1.94 (m, 2H), 1.98-2.15 (m, 6H), 2.19 (t, J=7.4 Hz, 2H), 2.24-2.34 (m, 3H), 2.36-3.04 (brs, 3H), 2.61 (t, J=7.8 Hz, 2H), 2.63-3.04 (brs, 10H), 3, 12-3.19 (m, 3H), 3.25-3.26 (m, 4H), 3.35-3.37 (m, 4H), 3.47-3.56 (m, 5H), 3.59-3.68 (m, 4H), 3.97 (d, J=4.0 Hz, 1H), 4.08-4.22 (m, 4H), 4.31 (s, 2H) , 7.1-7.18 (m, 3H), 7.24-7.27 (m, 2H);

MS (ESI) m/z 1577 [M+H] ⁺ .

Step 4: Preparation of Compound 15b.

Compound 15b (17 mg, 26%) was obtained according to the procedure described for the preparation of compound 15a in

- 18 041468 as a colorless liquid, except that 4-(p-tolyl)butyric acid (12 mg, 0.063 mmol), hydroxybenzotriazole (HOBt, 14 mg, 0.106 mmol), TBTU (34 mg, 0.106 mmol), diisopropylethylamine (0.028 ml, 0.159 mmol) and compound 14a (76 mg, 0.053 mmol) synthesized in step 3 above.

MS (ESI) m/z 1590 [M+H]+.

Step 4 Preparation of Compound 15c.

Compound 15c (36 mg, 51%) was prepared as described for compound 15a as a colorless liquid, except that 4-(p-iodophenyl)butyric acid (15 mg, 0.050 mmol), hydroxybenzotriazole (HOBT, 11 mg, 0.084 mmol), TBTU (27 mg, 0.084 mmol), diisopropylethylamine (0.022 ml, 0.126 mmol) and compound 14a (60 mg, 0.042 mmol) synthesized in step 3 above.

1H NMR (400 MHz, methanol-OO δ 1.44-1.48 (m, 63H), 1.51-1.57 (m, 4H), 1.58-1.70 (m, 3H), 1 .71-1.82 (m, 3H), 1.84-1.92 (m, 3H), 1.93-2.15 (m, 5H), 2.18 (t, J=7.6 Hz , 2H), 2.20-2.34 (m, 5H), 2.36-2.56 (brs, 3H), 2.58 (t, J=7.6 Hz, 2H), 2.61- 2.76(brs, 3H), 2.81(s, 2H), 2.86-3.09(brs, 5H), 3.11-3.18(m, 3H), 3.20-3, 26 (m, 3H), 3.35-3.39 (m, 2H), 3.42-3.48 (br, 2H), 3.53 (q, J=4.0 Hz, 2H), 3 .62 (s, 2H), 3.64-3.69 (m, 3H), 3.97 (d, J=3.6 Hz, 1H), 4.08 (s, 1H), 4.10- 4.23 (m, 4H), 4.31 (s, 2H), 6.32-6.36 (m, 1H), 6.99 (d, J=8.4 Hz, 2H), 7.60 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1702 [M+H] ⁺ .

Step 5: Preparation of Compound 2f.

Compound 15a (14 mg, 0.0089 mmol) synthesized in step 4 above was dissolved in 70% trifluoroacetic acid/dichloromethane (0.5 ml), then stirred at room temperature for 1 h. The reagent was concentrated under reduced pressure and the concentrate was separated by high efficiency liquid chromatography (HPLC) to give compound 2f (7 mg, 67%) as a white solid.

1H NMR (400 MHz, D ₂ O) δ 1.22-1.31 (m, 4H), 1.39 (n, J=7.4 Hz, 2H), 1.42-1.51 (m, 2H), 1.57-1.73 (m, 4H), 1.79 (n, J=7.6 Hz, 2H), 1.82-1.89 (m, 1H), 2.01-2 .09 (m, 1H), 2.13 (t, J=7.2 Hz, 2H), 2.39 (t, J=7.2 Hz, 2H), 2.51 (t, J=7, 4 Hz, 2H), 2.53-3.00 (br, 3H), 3.03 (t, J=6.8 Hz, 2H), 3.09-3.43 (m, 18H), 3, 47 (q, J=5.2 Hz, 2H), 3.48-3.60 (m, 6H), 3.61-3.91 (br, 5H), 3.96 (s, 2H), 4 .00-4.16 (m, 3H), 4.25 (s, 2H), 7.13-7.17 (m, 3H), 7.22-7.27 (m, 2H);

MS (ESI) m/z 1183 [M+H] ⁺ , 1181 [MH]'.

Step 5 Preparation of Compound 2g.

Compound 2g (10 mg, 36%) is prepared as described for compound 2f as a white solid, except that compound 15b (37 mg, 0.023 mmol) synthesized in step 4 above and 70% trifluoroacetic acid/ dichloromethane (0.5 ml).

1H NMR (400 MHz, D ₂ O) δ 1.14-1.26 (m, 4H), 1.35 (n, J=6.8 Hz, 2H), 1.39-1.48 (m, 2H), 1.50-1.63 (m, 3H), 1.66-1.69 (m, 1H), 1.72 (n, J=7.2 Hz, 2H), 1.82 (n , J=7.2 Hz, 1H), 2.01 (n, J=7.0 Hz, 1H), 2.08 (t, J=7.2 Hz, 2H), 2.15 (s, 3H ), 2.35 (t, J=7.2 Hz, 2H), 2.43 (t, J=7.4 Hz, 2H), 2.66-2.98 (br, 2H), 2.99 (t, J=6.6Hz, 2H), 3.01-3.40(m, 18H), 3.44(kv, J=5.0Hz, 2H), 3.48-3.56( m, 5H), 3.57-3.88 (br, 6H), 3.92 (s, 2H), 3.98-4.12 (m, 4H), 4.21 (s, 2H), 7 .01 (d, J=8.0 Hz, 2H), 7.04 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1198 [M+H] ⁺ , 1196 [MH]'.

Step 5: Preparation of Compound 2h.

Compound 2h (13 mg, 54%) is obtained according to the procedure described for the preparation of compound 15a as a white solid, except that compound 15c (30 mg, 0.018 mmol) synthesized in step 4 above and 70% trifluoroacetic acid/ dichloromethane (0.5 ml).

1H NMR (400 MHz, D ₂ O) δ 1.18-1.30 (m, 4H), 1.37 (n, J=6.8 Hz, 2H), 1.41-1.52 (m, 2H), 1.54-1.67 (m, 3H), 1.70-1.74 (m, 1H), 1.78 (t, J=7.4 Hz, 2H), 1.81-1 .90 (m, 1H), 1.98-2.07 (m, 1H), 2.11 (t, J=7.2 Hz, 2H), 2.39 (t, J=7.2 Hz, 2H), 2.48 (t, J=7.4 Hz, 2H), 3.00 (t, J=6.8 Hz, 2H), 3.02-3.26 (m, 17H), 3, 29-3.35(m, 2H), 3.48(q, J=4.4Hz, 2H), 3.52-3.58(m, 6H), 3.60-3.93(brs, 7H), 3.97 (s, 2H), 4.03-4.17 (m, 3H), 4.25 (s, 2H), 6.95 (d, J=8.4 Hz, 2H), 7.59 (d, J=8.4 Hz, 2H);

MS (ESI) m/z 1309 [M+H] ⁺ , 1307 [MH]'.

- 19 041468

Example 6 Preparation of compound 2i.

Step 1: Preparation of Compound 9b.

Compound 3b (691 mg, 1.15 mmol) synthesized in Example 1 was dissolved in dichloromethane (10 ml), N,N'-dicyclohexylcarbodiimide (DCC, 3710 mg, 1.79 mmol) was slowly added thereto at room temperature. 2-(2-azidoethoxy)acetic acid 8b (N ₃ -CH ₂ CH ₂ OCH2COOH, 200 mg, 1.38 mmol) was added thereto, followed by stirring for 1 h. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (40% ethyl acetate/n-hexane) to give compound 9b (670 mg, 80%) as a colorless liquid.

¹ H NMR (400 MHz, methanol-d4) δ 1.44-1.51 (m, 36H), 1.52-1.67 (m, 4H), 1.71-1.84 (m, 2H) , 2.00-2.09 (m, 1H), 2.25-2.38 (m, 2H), 3.36 (q, J=7.5 Hz, 2H), 3.41-3.45 (m, 2H), 3.66 (t, J=5.0 Hz, 1H), 3.71 (t, J=5.0 Hz, 1H), 3.97 (d, J=0.8 Hz , 1H), 4.11 (d, J=1.2 Hz, 1H), 4.12-4.23 (m, 3H), 4.32 (s, 2H), 6.34 (n, J= 4.2 Hz, 2H);

MS (ESI) m/z 729 [M+H]+.

Step 2: Preparation of Compound 10b.

Compound 9b (650 mg, 0.892 mmol) synthesized in Step 1 above was dissolved in ethanol (20 ml), 10% palladium-carbon (95 mg) was added thereto, followed by stirring for 12 hours under hydrogen. The reaction solution is filtered, washed with ethanol, and concentrated under reduced pressure. The concentrate was separated by column chromatography (2% methanol/dichloromethane, NH silica gel) to give compound 10b (573 mg, 91%) as a colorless liquid.

1H NMR (400 MHz, methanol-d4) δ 1.37-1.42 (m, 2H), 1.44-1.49 (m, 36H), 1.51-1.67 (m, 4H), 1.71-1.84 (m, 2H), 1.99-2.09 (m, 1H), 2.29-2.34 (m, 2H), 2.84 (n, J=5.2 Hz, 2H), 3.35-3.40 (m, 1H), 3.54 (t, J=5.4 Hz, 1H), 3.59 (t, J=5.4 Hz, 1H), 3.98 (d, J=0.8 Hz, 1H), 4.07 (s, 1H), 4.09-4.21 (m, 2H), 4.31 (s, 2H);

MS (ESI) m/z 703 [M+H] ⁺ .

Step 3: Preparation of Compound 11b.

DOTA-tris(tBu) ether (82 mg, 0.143 mmol) was dissolved in dichloromethane (2.0 ml), hydroxybenzotriazole (HOBt, 32 mg, 0.239 mmol), TBTU (77 mg, 0.239 mmol) and diisopropylethylamine (0.062 ml) were added thereto. , 0.358 mmol), then stirred at room temperature for 10 minutes. Compound 10b (84 mg, 0.120 mmol) synthesized in Step 2 above, dissolved in dichloromethane (2.0 ml) was added thereto, followed by stirring at room temperature for 1 hour. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (2% methanol/dichloromethane) to give compound 11b (65 mg, 43%) as a colorless liquid.

1H NMR (400 MHz, methanol^) δ 1.44-1.49 (m, 63H), 1.51-1.54 (m, 2H), 1.57-1.65 (m, 2H), 1 .75-1.84(m, 2H), 2.04-2.09(m, 2H), 2.30-2.34(m, 2H), 2.81-3.25(brs, 18H) , 3.35-3.54 (brs 7H), 3.55 (t, J=5.4 Hz, 2H), 3.61 (t, J=5.2 Hz, 2H), 3.98 (s , 2H), 4.06 (s, 1H), 4.10-4.22 (m, 2H), 4.29 (s, 2H);

MS (ESI) m/z 1258 [M+H] ⁺ .

- 20 041468

Step 4 Preparation of Compound 2i.

Compound 11b (55 mg, 0.044 mmol) synthesized in step 3 above was dissolved in 70% trifluoroacetic acid/dichloromethane (0.5 ml), then stirred at room temperature for 4 h. The reagent was concentrated under reduced pressure and the concentrate was separated high performance liquid chromatography (HPLC) to give compound 2i (17 mg, 45%) as a white solid.

Ή NMR (400 MHz, D2O) δ 1.20-1.34 (m, 2H), 1.42-1.54 (m, 2H), 1.56-1.65 (m, 1H), 1, 70-1.77 (m, 1H), 1.86 (n, J=7.4 Hz, 1H), 2.05 (n, J=7.2 Hz, 1H), 2.39 (t, J =7.2Hz, 2H), 2.84-3.49(m, 20H), 3.51(t, J=5.0Hz, 1H), 3.55(t, J=4.0Hz , 1H), 3.58-3.62 (brs, 4H), 3.63-3.95 (brs, 3H), 4.00 (s, 2H), 4.05 (s, 1H), 4, 07-4.17 (m, 2H), 4.29 (s, 2H);

MS (ESI) m/z 865 [M+H]+, 863 [MH] ^- .

Example 7 Preparation of Compounds 2j and 2k

Stage 1. Preparation of compound 12b.

Lysine (Fmoc-Lys(Z)-OH, 386 mg, 0.768 mmol) is dissolved in dichloromethane (10 ml), hydroxybenzotriazole (HOBt, 173 mg, 1.28 mmol), TBTU (411 mg, 1.28 mmol) is added thereto and diisopropylethylamine (0.335 ml, 1.92 mmol), then stirred at room temperature for 10 minutes. Compound 10b (450 mg, 0.640 mmol) synthesized in Step 2 of Example 6 was added thereto, dissolved in dichloromethane (5.0 ml), followed by stirring at room temperature for 1 hour. Water was added to the reagent, and then the organic compound was re-extracted 3 times with dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (2% methanol/dichloromethane). Dichloromethane (15 ml) was added to the resulting compound, piperidine (0.050 ml, 0.505 mmol) was added thereto, followed by stirring at room temperature for 24 hours. The reagent was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane, NH silica gel) to give compound 12b (380 mg, 61%) as a colorless liquid.

1H NMR (400 MHz, methanol^) δ 1.33-1.41 (m, 4H), 1.44-1.48 (m, 36H), 1.51-1.72 (m, 8H), 1 .74-1.84 (m, 2H), 1.99-2.08 (m, 1H), 2.24-2.38 (m, 2H), 3.11 (t, J=6.8 Hz , 2H), 3.24-3.28 (m, 1H), 3.34-3.46 (m, 3H), 3.55 (t, J=5.4 Hz, 1H), 3.60 ( t, J=5.4 Hz, 1H), 3.96 (s, 1H), 4.05 (d, J=1.2 Hz, 1H), 4.16-4.22 (m, 3H), 4.29 (s, 1H), 5.05 (s, 2H), 7.26-7.32 (m, 1H), 7.33-7.38 (m, 4H);

MS (ESI) m/z 966 [M+H] ⁺ .

Step 2: Preparation of compound 13b.

DOTA-tris(tBu) ether (271 mg, 0.472 mmol) was dissolved in dichloromethane (10 ml), hydroxybenzotriazole (HOBt, 106 mg, 0.787 mmol), TBTU (253 mg, 0.787 mmol) and diisopropylethylamine (0.206 ml, 1 .18 mmol), then stirred at room temperature for 10 min. Compound 12b (380 mg, 0.394 mmol) synthesized in Step 1 above, dissolved in dichloromethane (5.0 ml) was added thereto, followed by stirring at room temperature for 1 hour. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (3% methanol/dichloromethane) to give compound 13b (487 mg, 81%) as a colorless liquid.

- 21 041468 ¹ H NMR (400 MHz, methanol-d ₄ ) δ 1.44-1.49 (m, 63H), 1.51-1.53 (m, 2H), 1.59-1.66 ( m, 4H), 1.75-1.84 (m, 4H), 2.01-2.09 (m, 3H), 2.10-2.26 (br, 4H), 2.27-2, 34 (m, 3H), 2.38-2.94 (br, 12H), 2.95-3.21 (m, 6H), 3.23-3.27 (m, 2H), 3.32- 3.64 (m, 8H), 3.99-4.08 (m, 2H), 4.11-4.22 (m, 3H), 4.30 (s, 2H), 5.06 (s, 2H), 7.28-7.31 (m, 1H), 7.33-7.34 (m, 4H);

MS (ESI) m/z 1520 [M+H]+.

Step 3: Preparation of Compound 14b.

Compound 13b (467 mg, 0.307 mmol) synthesized in Step 2 above was dissolved in ethanol (20 ml), 10% palladium-carbon (33 mg) was added thereto, followed by stirring for 2 hours under hydrogen. The reaction solution is filtered, washed with ethanol, and concentrated under reduced pressure. The concentrate was separated by column chromatography (2% methanol/dichloromethane, NH silica gel) to give compound 14b (366 mg, 86%) as a colorless liquid.

¹ H NMR (400 MHz, methanol^) δ 1.44-1.49 (m, 63H), 1.51-1.67 (m, 8H), 1.74-1.84 (m, 3H), 1.89-2.00(brs, 1H), 2.01-2.08(m, 2H), 2.09-2.26(brs, 5H), 2.29-2.34(m, 2H ), 2.36-2.62(brs, 5H), 2.63-2.68(m, 2H), 2.70-3.22(brs, 10H), 3.26(t, J=7 .6 Hz, 2H), 3.39 (t, J=7.2 Hz, 2H), 3.42-3.73 (m, 6H), 3.95-4.06 (m, 2H), 4 .08-4.21 (m, 4H), 4.32 (s, 2H);

MS (ESI) m/z 1386 [M+H] ⁺ .

Step 4 Preparation of Compound 15d.

4-(p-Tolyl)butyric acid (11 mg, 0.059 mmol) was dissolved in dichloromethane (1.0 ml), hydroxybenzotriazole (HOBt, 13 mg, 0.098 mmol), TBTU (32 mg, 0.098 mmol) and diisopropylethylamine ( 0.026 ml, 0.147 mmol), then stirred at room temperature for 10 minutes. Compound 14b (68 mg, 0.049 mmol) synthesized in Step 3 above, dissolved in dichloromethane (1.0 ml) was added thereto, followed by stirring at room temperature for 2 hours. Water was added to the reagent, and then the organic compound was re-extracted 3 times using dichloromethane. The collected organic solvent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (4% methanol/dichloromethane) to give compound 15d (60 mg, 42%) as a colorless liquid.

Step 4: Preparation of Compound 15e.

Compound 15e (36 mg, 51%) was prepared as described for compound 15a as a colorless liquid, except that 4-(p-iodophenyl)butyric acid (32 mg, 0.104 mmol), hydroxybenzotriazole (HOBT, 23 mg, 0.173 mmol), TBTU (56 mg, 0.173 mmol), diisopropylethylamine (0.045 ml, 0.260 mmol) and compound 14b (120 mg, 0.087 mmol) synthesized in step 3 above.

¹ H NMR (400 MHz, CDCl3) δ 0.82-0.96 (m, 2H), 0.97-1.14 (m, 2H), 1.16-1.37 (m, 6H), 1 .42-1.50 (m, 63H), 1.53-1.69 (m, 2H), 1.71-1.96 (m, 5H), 1.99-2.10 (m, 3H ), 2.11-2.38(m, 11H), 2.41-2.68(m, 6H), 2.81(s, 3H), 2.82-3.11(brs, 7H), 3.16-3.33 (m, 5H), 3.35-3.61 (m, 5H), 3.63-3.75 (m, 2H), 3.90 (s, 2H), 4, 10 (d, J=3.2 Hz, 1H), 4.24-4.35 (m, 3H), 5.60 (q, J=7.6 Hz, 1H), 6.97 (d, J =8.0 Hz, 2H), 7.55 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1658 [M+H] ⁺ .

Step 5 Preparation of Compound 2j.

Compound 15d (24 mg, 0.016 mmol) synthesized in step 4 above was dissolved in 70% trifluoroacetic acid/dichloromethane (0.5 ml), then stirred at room temperature for 1 h. The reagent was concentrated under reduced pressure and the concentrate was separated by high efficiency liquid chromatography (HPLC) to give compound 2j (6.7 mg, 37%) as a white solid.

¹ H NMR (400 MHz, D2O) δ 1.16-1.28 (m, 4H), 1.35-1.48 (m, 4H), 1.52-1.64 (m, 3H), 1 .65-1.70 (m, 1H), 1.74 (t, J=7.6 Hz, 2H), 1.79-1.87 (m, 1H), 2.00-2.05 (m , 1H), 2.10 (t, J=7.0 Hz, 2H), 2.16 (s, 3H), 2.36 (t, J=6.6 Hz, 2H), 2.44 (t , J=7.2 Hz, 2H), 2.82-3.10 (m, 8H), 3.11-3.27 (m, 9H), 3.28-3.38 (m, 4H), 3.39-3.49 (m, 4H), 3.50-3.80 (m, 7H), 3.93 (s, 2H), 3.98-4.12 (m, 3H), 4, 19 (s, 2H), 7.02 (d, J=8.0 Hz, 2H), 7.06 (d, J=7.2 Hz, 2H);

MS (ESI) m/z 1153 [M+H] ⁺ , 1151 [MH] ^- .

Stage 5. Obtaining connection 2k.

Compound 2k (4.0 mg, 53%) was prepared as described for compound 2j as a white solid, except compound 15e (10 mg, 0.0060 mmol) synthesized in step 4 above was used.

¹ H NMR (400 MHz, D ₂ O) δ 1.15-1.27 (m, 4H), 1.28-1.36 (m, 2H), 1.38-1.48 (m, 2H) , 1.54-1.64 (m, 3H), 1.66-1.68 (m, 1H), 1.74 (p, J=7.4 Hz, 2H), 1.79-1.87 (m, 1H), 1.98-2.03 (m, 1H), 2.08 (t, J=7.0 Hz, 2H), 2.36 (t, J=7.4 Hz, 2H) , 2.43 (t, J=7.2 Hz, 2H), 2.71-2.87 (brs, 3H), 2.96 (t, J=6.6 Hz, 2H), 3.03- 3.33(m, 17H), 3.38-3.49(m, 3H), 3.52-3.89(brs, 6H), 3.94(d, J=4.4Hz, 2H) , 4.00-4.14 (m, 4H), 4.19 (s, 2H), 6.90 (d, J=8.4 Hz, 2H), 7.54 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1266 [M+H] ⁺ , 1264 [MH] ^- .

- 22 041468

Example 8 Preparation of Compound 21

dichloromethane (10 ml), add

Stage 1. Preparation of compound 16.

Fmoc-Gly-OH (0.49 g, 1.66 mmol) is dissolved in

K,K'-dicyclohexylcarbodiimide (0.34 g, 1.66 mmol) then stirred at 0° C. for 10 min. Compound 3b (0.50 g, 0.83 mmol) dissolved in dichloromethane (10 ml) was slowly added to the reaction mixture, then stirred at 0°C for 1.5 h. The reaction mixture was filtered, washed with dichloromethane several times and the solvent is removed under reduced pressure. The concentrate was separated by column chromatography (35%-50% ethyl acetate/n-hexane) to give compound 16 (0.45 g, 61%).

¹ H NMR (400 MHz, CDCl3) δ 1.05-1.21 (m, 2H), 1.38 (s, 9H), 1.40 (s, 9H), 1.43 (s, 9H), 1.45 (s, 9H), 1.56-1.62 (m, 2H), 1.64-1.71 (m, 2H), 1.75-1.86 (m, 2H), 1. 88-1.94 (m, 2H), 2.19-2.35 (m, 2H), 3.16-3.29, 3.88-4.02 (m, 2H), 4.19-4 .28 (m, 2H), 4.29-4.40 (m, 3H), 5.23 (dt, J=64.4, 12.0 Hz, 2H), 5.96 (dt, J=105 .2, 4.4 Hz, 1H), 7.27-7.30 (m, 2H), 7.37 (t, J=13.4 Hz, 2H), 7.55 (d, J=7, 2 Hz, 2H), 7.74 (d, J=7.2 Hz, 2H);

MS (ESI) m/z 881 [M+H]+.

Step 2 Preparation of Compound 10c.

Compound 16 (0.40 g, 0.45 mmol) was dissolved in dichloromethane (10 ml), piperidine (0.1 ml, 1.01 mmol) was added thereto, then stirred at room temperature for 1.5 h. concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 10c (0.19 g, 63%).

1H NMR (400 MHz, CDCl3) δ 1.41 (s, 9H), 1.43 (s, 9H), 1.46 (s, 9H), 1.47 (s, 9H) 1.53-1, 64 (m, 2H), 1.75-1.89 (m, 2H), 2.02-2.11 (m, 2H), 2.24-2.38 (m, 2H), 3.23 ( t, J=7.4 Hz, 2H), 3.63 (s, 2H), 3.69 (s, 2H), 3.84-3.97 (m, 2H), 4.22-4.37 (m, 2H), 5.46-5.84 (m, 2H);

MS (ESI) m/z 659 [M+H] ⁺ .

Step 3: Preparation of compound 11c.

DOTA-tpuc(IBu) ester (31 mg, 55 µmol) is dissolved in dichloromethane (0.6 ml), hydroxybenzotriazole (HOBT, 9 mg, 68 µmol), TBTU (22 mg, 68 µmol) and diisopropylethylamine (16 µL) are added thereto. , 91 µmol), then stirred at room temperature for 10 min. Compound 10c (30 mg, 45.5 µmol) dissolved in dichloromethane (0.3 ml) was added thereto, followed by stirring at room temperature for 1 hour. The reaction was quenched by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 mlx3). The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 11c (47.4 mg, 86%).

1H NMR (400 MHz, CDCl3) δ 1.46 (s, 9H), 1.47 (s, 27H), 1.48 (s, 27H), 1.68 (s, 6H), 1.73-1 .80 (m, 1H), 1.78-1.90 (m, 1H), 2.04-2.15 (m, 3H), 2.20-2.48 (m, 6H), 2.83 (s, 9H), 3.00 (brs, 3H), 3.27 (t, J=7.4 Hz, 2H), 3.32-3.41 (m, 2H), 3.44-3, 54 (m, 2H), 3.91-4.14 (m, 3H), 4.20-4.40 (m, 2H), 5.25-5.34 (m, 1H), 5.46 ( dd, J=7.4 Hz, 1H);

MS (ESI) m/z 1213 [M+H] ⁺ .

Step 4 Preparation of Compound 21

Compound 11c (40 mg, 32.96 µmol) was added to 70% trifluoroacetic acid/dichloromethane (1 ml), then stirred for 5 h. centrifuges. Compound 21 (15 mg, 55%) was then obtained by high performance liquid chromatography (HPLC).

1H NMR (400 MHz, D2O) δ 1.29-1.38 (m, H), 1.46 (brb, 1H), 1.54-1.62 (m, 2H), 1.63-1, 70 (m, 1H), 1.75-1.82 (m, 1H), 1.83-1.93 (m, 1H), 2.05-2.13 (m, 1H), 2.42 ( t, J=7.2 Hz, 2H), 2.70-3.89 (br, 22H), 3.94 (s, 1H), 3.99 (s, 1H), 4.04 (s, 2H ), 4.07-4.12 (1H), 4.12-4.15 (m, 2H), 4.18 (t, J=4.4 Hz, 2H);

MS (ESI) m/z 821 [M+H] ⁺ .

- 23 041468

Example 9 Preparation of Compound 2m.

Stage 1. Preparation of compound 17.

Fmoc-Lys(Z)-OH (183 mg, 0.36 mmol) is dissolved in dichloromethane (1 ml), hydroxybenzotriazole (HOBt, 62 mg, 0.46 mmol), TBTU (146 mg, 0.46 mmol) is added thereto and diisopropylethylamine (106 μL, 0.61 mmol), then stirred at room temperature for 10 minutes. Compound 10c (0.20 g, 0.30 mmol) dissolved in dichloromethane (1 ml) was added thereto, then stirred for 4 h. The reaction was stopped by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 mlx3) . The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 17 (268 mg, 77%) as a white solid.

Ή NMR (400 MHz, CDCl3) δ 1.19-1.29 (m, 2H), 1.37-1.46 (m, 36H), 1.49-1.55 (m, 2H), 1, 57-1.63 (m, 2H), 1.68 (s, 3H), 1.77-1.84 (m, 2H), 2.00-2.10 (m, 2H), 2.22- 2.35 (m, 2H), 3.10-3.23 (m, 3H), 3.79-3.99 (m, 3H), 4.13 (s, 1H), 4.18-4, 26 (m, 2H), 4.32-4.42 (m, 3H), 5.05 (s, 2H), 5.37-5.87 (m, 3H), 7.02 (d, J= 22 Hz, 1H), 7.23-7.40 (m, 8H), 7.54-7.60 (m, 2H), 7.73 (kv, J=6.1 Hz, 2H);

MS (ESI) m/z 1165 [M+Na]+.

Step 2 Preparation of Compound 12c.

Compound 17 (250 mg, 0.22 mmol) was dissolved in CH ₂ Cl ₂ (1 ml), piperidine (64.80 μl, 0.66 mmol) was added thereto, then stirred at room temperature for 5 hours. After removal of the solvent from the reaction mixture under reduced pressure, the concentrate was subjected to column chromatography to obtain compound 12c (170 mg, 84%) as a colorless liquid.

Ή NMR (400 MHz, CDClg) δ 1.24-1.36 (m, 2H), 1.45 (s, 9H), 1.47 (s, 9H), 1.48 (s, 18H), 1 .52-1.59 (m, 4H), 1.71 (s, 4H), 1.77-1.89 (m, 3H), 2.04-2.13 (m, 1H), 2.25 -2.39 (m, 2H), 3.15-3.25 (m, 2H), 3.26-3.34 (m, 1H), 3.85-4.05 (m, 2H), 4 .12-4.20 (m, 2H), 4.26-4.41 (m, 2H), 5.06-5.16 (m, 3H), 5.51 (ddd, J=49.3, 29.4, 8.1 Hz, 2H), 7.37 (d, J=4.4 Hz, 4H), 7.92 (s, 1H);

MS (ESI) m/z 922 [M+H] ⁺ .

Step 3: Preparation of compound 13c.

DOTA-tpuc(IBu) ester (99 mg, 0.119 mmol) was dissolved in dichloromethane (0.5 ml), hydroxybenzotriazole (HOBt, 23 mg, 0.261 mmol), TBTU (56 mg, 0.261 mmol) and diisopropylethylamine (30 μl) were added thereto. , 347 µmol), then stirred at room temperature for 10 min. Compound 12c (160 mg, 0.174 mmol) dissolved in dichloromethane (1.6 ml) was added thereto, then stirred for 1 h. The reaction was stopped by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 mlx3). The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 13c (180 mg, 72%) as a colorless liquid.

Ή NMR (400 MHz, CDCF) δ 1.28-1.33 (m, 3H), 1.45 (s, 9H), 1.46 (s, 9H), 1.47 (s, 9H), 1 .47(s, 9H), 1.48(s, 18H), 1.49(s, 9H), 1.79(s, 8H), 2.04-2.15(m, 5H), 2, 25-2.41(m, 5H), 2.59(brs, 3H), 2.83(brs, 4H), 2.95(brs, 3H), 3.18-3.28(m, 5H) , 3.47 (ws, 4H), 3.92 (dd, J=44.0, 18.4 Hz, 2H), 4.04-4.24 (m, 2H), 4.33-4.39 (m, 3H), 5.10 (d, J=3.2 Hz, 2H), 7.15 (d, J=18.4 Hz, 1H), 7.30-7.37 (m, 4H) , 7.46 (s, 1H);

MS (ESI) m/z 1498 [M+Na]+.

- 24 041468

Step 4 Preparation of Compound 14c.

Palladium (10% palladium on carbon, 6 mg, 5.8 µmol) is placed in a round bottom flask, which is sealed with a septum and filled with hydrogen under vacuum. Compound 13c (170 mg, 115 μmol) dissolved in methanol (2 ml) was charged into a reaction vessel, then stirred at room temperature for 15 h. The reagent was filtered with celite, concentrated under reduced pressure, and the concentrate was separated by NH column chromatography -silica gel (0-1% methanol/dichloromethane) to give compound 14c (117 mg, 76%) as a colorless liquid.

1H NMR (400 MHz, CDC13) δ 1.14-1.28 (m, 3H), 1.36 (s, 18H), 1.41 (s, 18H), 1.44 (s, 27H), 1 .53-1.62 (m, 4H), 1.68-1.83 (m, 5H), 1.97-2.09 (m, 4H), 2.10-2.37 (m, 7H) , 2.40-2.69 (m, 6H), 2.70-3.10 (m, 7H), 3.15-3.26 (m, 2H), 3.31-3.65 (m, 4H), 3.78-3.95(m, 2H), 4.01-4.13(m, 2H), 4.21-4.35(m, 3H), 5.37-5.55( m, 2H), 7.17 (d, J=28.8 Hz, 1H), 7.52 (brs, 1H);

MS (ESI) m/z 1365 [M+Na]+.

Step 5: Preparation of Compound 15f.

4-(p-Tolyl)butyric acid (16 mg, 89 µmol) is dissolved in dichloromethane (1 ml), hydroxybenzotriazole (HOBt, 15 mg, 112 µmol), TBTU (36 mg, 112 µmol) and DIEA (26 µl) are added thereto , 149 µmol), then stirred at room temperature for 10 min. Compound 14c (100 mg, 75 µmol) dissolved in dichloromethane (2 ml) was added thereto, followed by stirring for 1.5 h. The reaction was quenched by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 mlx3). The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography to give compound 15f (69 mg, 86%) as a colorless liquid.

1H NMR (400 MHz, CDC13) δ 1.26-1.32 (m, 2H), 1.35-1.40 (m, 27H), 1.42-1.47 (m, 36H), 1, 52-1.59 (m, 4H), 1.79-1.96 (m, 6H), 2.00-2.08 (m, 4H), 2.10-2.20 (m, 2H), 2.21-2.41 (m, 14H), 2.54-2.62 (m, 6H), 2.78 (s, 3H), 2.83-2.95 (m, 2H), 3, 20 (s, 3H), 3.28-3.62 (m, 4H), 3.73-3.88 (m, 2H), 3.93-4.03 (m, 2H), 4.13- 4.25(m, 2H), 4.27-4.33(m, 2H), 5.35-5.61(m, 2H), 7.05(d, J=1.6Hz, 4H) .

MS (ESI) m/z 1524 [M+Na] ⁺ .

Step 6. Preparation of compound 2m.

Compound 15f (86 mg, 57 μmol) obtained in step 5 above was added to 70% trifluoroacetic acid/dichloromethane (1 ml), then stirred for 6 h. After pouring the reaction mixture into diethyl ether (40 ml) to give a precipitate , it is separated using a centrifuge. The mixture was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound 2m (44 mg, 69%) as a white solid.

1H NMR (400 MHz, D2O) δ 1.36-1.41 (m, 4H), 1.46-1.53 (m, 3H), 1.54-1.59 (m, 2H), 1, 66-1.75 (m, 2H), 1.77-1.88 (m, 4H), 1.90-1.99 (m, 1H), 2.10-2.16 (m, 1H), 2.21 (t, J=7.2 Hz, 2H), 2.27 (s, 3H), 2.48 (t, J=7.6 Hz, 2H), 2.53 (t, J=7 .6 Hz, 2H), 2.93-3.56 (br, 20H), 3.71 (br, 3H), 3.87 (s, 1H), 3.91 (s, 1H), 3.98 (s, 1H), 4.04 (t, J=8.6 Hz, 3H), 4.12 (s, 1H), 4.13-4.19 (m, 2H), 4.22-4, 26 (m, 1H), 4.30 (br, 1H), 7.15 (q, J=7.9 Hz, 4H);

MS (ESI) m/z 1109 [MH] ^- .

Example 10 Preparation of Compound 19

R ₄ I

19a, R ₄ = -CH ₂ CO ₂ tBu

19b, R ₄ = -CH ₂ CH ₂ CO ₂ tBu

Preparation of compound 19a.

Compound 18 (500 mg, 2.87 mmol) was dissolved in dichloromethane (5 ml) and cooled to 0° C., triethylamine (0.6 ml, 4.31 mmol) was added thereto. tert-Butyl bromoacetate (620 mg, 3.16 mmol) dissolved in dichloromethane (5 ml) was slowly added thereto, followed by stirring at room temperature for 18 hours. The reaction was quenched by adding water (10 ml) and the organic compound was extracted using dichloromethane ( 10 mlx2). The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (2% methanol/dichloromethane) to give compound 19a (0.46 g, 55%).

1H NMR (400 MHz, CDC13) δ 1.45 (s, 9H), 2.78 (t, J=4.8 Hz, 2H), 3.31 (s, 2H), 3.38 (t, J =5.6 Hz, 2H), 3.68-3.59 (m, 9H);

¹³ C NMR (100 MHz, CDC13) δ 28.1, 48.7, 50.6, 51.7, 56.6, 76.7, 77.0, 77.3, 81.0, 171.5;

MS (ESI) m/z 289 [M+H] ⁺ .

Preparation of compound 19b.

Compound 18 (300 mg, 1.72 mmol) was dissolved in ethanol (10 ml) and cooled to 0° C., tert-butyl acrylate (220 mg, 1.72 mmol) was slowly added thereto. After gradually raising the temperature of the mixture to room temperature, the mixture was stirred for 18 hours. The organic solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 19b (380 mg, 73%).

- 25 041468

1H NMR (400 MHz, CDCI3) δ 1.43 (s, 9H), 2.42 (t, J=6.4 Hz, 2H), 2.79 (t, J=5.6 Hz, 2H), 2.85 (t, J=6.4 Hz, 2H), 3.38 (t, J=4.8 Hz, 2H), 3.58 (t, J=5.2 Hz, 2H), 3. 62-3.68 (m, 6H);

¹³ C NMR (100 MHz, CDCI3) δ 28.1, 35.9, 45.2, 49.1, 50.7, 76.7, 77.0, 77.3, 80.4, 172.0;

MS (ESI) m/z 303 [M+H]+.

Example 11 Preparation of Compounds 2n and 2o

Step 1: Preparation of compound 21a.

Compound 20 (180 mg, 0.36 mmol) was dissolved in dichloromethane (5 ml) and cooled to 0°C, N,N'-dicyclohexylcarbodiimide (DCC, 83 mg, 0.40 mmol) and compound 19a (110 mg , 0.36 mmol), then stirred at room temperature for 1 hour. The organic layer was filtered several times and the solvent was removed under reduced pressure. The concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 21a (250 mg, 91%).

1H NMR (400 MHz, CDC13) δ 1.43 (s, 9H), 1.44 (s, 9H), 1.45 (s, 9H), 1.46 (s, 9H), 1.79-1 .86 (m, 1H), 2.01-2.20 (m, 3H), 2.26-2.36 (m, 3H), 2.44-2.55 (m, 1H), 2.77 (w, 1H), 3.40 (t, J=6.4 Hz, 2H), 3.56-3.74 (m, 9H), 4.03 (dd, J=44.0, 17.2 Hz, 2H), 4.22-4.35 (m, 2H);

MS (ESI) m/z 759 [M+H] ⁺ .

Step 1: Preparation of Compound 21b.

Compound 21b (275 mg, 87%) was prepared as described for compound 21a, except that compound 20 (200 mg, 0.41 mmol), N,N'-dicyclohexylcarbodiimide (DCC, 102 mg, 0 .49 mmol) and compound 19b (200 mg, 0.41 mmol).

MS (ESI) m/z 773 [M+H] ⁺ .

Step 2: Preparation of compound 22a.

Compound 21a (200 mg, 0.26 mmol) was dissolved in methanol (8 ml), palladium (10% palladium on charcoal, 13 mg, 13 μmol) was added thereto. The reaction flask was filled with hydrogen, then stirred at room temperature for 1 hour. After passing the reaction product through Celite, the solvent was removed under reduced pressure and the concentrate was separated by column chromatography (8% methanol/dichloromethane) to give compound 22a (120 mg, 63%) .

1H NMR (400 MHz, methanol^) δ 1.45 (s, 9H), 1.47-1.49 (m, 27H), 1.76-1.94 (m, 2H), 2.01-2 .13 (m, 2H), 2.27-2.39 (m, 3H), 2.55-2.60 (m, 2H), 2.79-2.81 (m, 2H), 3.35 (s, 1H), 3.52 (t, J=5.2 Hz, 2H), 3.55-3.65 (m, 8H), 4.04 (dd, J=17.6, 4.8 Hz, 1H), 4.14-4.22 (m, 3H);

MS (ESI) m/z 733 [M+H] ⁺ .

Step 2: Preparation of compound 22b.

Compound 22b (210 mg, 90%) was prepared as described for compound 22a, except that compound 21b (240 mg, 0.32 mmol) and palladium (10% palladium on carbon, 17 mg, 16 µmol) were used. ).

1H NMR (400 MHz, methanol^) δ 1.45 (s, 18H), 1.48 (s, 18H), 1.59-1.63 (m, 1H), 1.68-1.75 (m , 2H), 1.79-1.91 (m, 4H), 2.03-2.14 (m, 2H), 2.30-2.36 (m, 2H), 2.48-2.62 (m, 3H), 2.80-2.82 (m, 2H), 3.43-3.70 (m, 12H), 4.14-4.20 (m, 2H);

MS (ESI) m/z 747 [M+H ^]+ .

Step 3: Preparation of compound 23a.

DOTA-tris(tBu) ether (38 mg, 65 µmol) was dissolved in dichloromethane (7 ml), hydro

- 26 041468 xybenzotriazole (HOBt, 110 mg, 82 µmol), TBTU (26 mg, 82 µmol) and diisopropylethylamine (19 µl, 0.11 mmol), then stirred for 10 minutes. Compound 22a (40 mg, 55 µmol) dissolved in dichloromethane (3 ml) was added thereto, followed by stirring for 1 h. The reaction was quenched by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 mlx2). The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to give compound 23a (60 mg, 70%) as a white solid.

1H NMR (400 MHz, methanol^) δ 1.40-1.57 (m, 63H), 1.76-2.87 (m, 26H), 3.36-3.63 (m, 17H), 3 .74 (s, 1H), 4.04 (d, J=8.0 Hz, 2H), 4.14-4.24 (m, 3H), 6.34-6.38 (m, 1H);

MS (ESI) m/z 1288 [M+H]+.

Step 3: Preparation of Compound 23b.

Compound 23b (60 mg, 68%) was prepared as described for compound 23a, except that DOTA-tpuc(IBu) ester (46 mg, 80 μmol), hydroxybenzotriazole (HOBT, 14 mg, 0.10 mmol), TBTU (32 mg, 0.10 mmol), diisopropylethylamine (24 μl, 0.13 mmol) and compound 22b (50 mg, 67 μmol).

¹ H NMR (400 MHz, methanol^) δ 1.09-1.24 (m, 12H), 1.29-1.43 (m, 10H), 1.45-1.53 (m, 41H), 1.57-1.63 (m, 4H), 1.66-1.76 (m, 7H), 1.80-1.91 (m, 8H), 2.03-2.12 (m, 2H ), 2.25-2.37 (m, 2H), 2.44-2.64 (m, 4H), 2.82 (s, 9H), 3.35-3.49 (m, 5H), 3.52-3.69 (m, 7H), 4.14-4.22 (m, 2H);

MS (ESI) m/z 1302 [M+H] ⁺ .

Step 4: Preparation of compound 2n.

Compound 23a (45 mg, 35 µmol) was added to 70% trifluoroacetic acid/dichloromethane (1 ml), then stirred for 7 h. After pouring the reaction mixture into diethyl ether (20 ml) to give a precipitate, it was separated using a centrifuge . The mixture was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound 2n (22 mg, 70%) as a solid.

1H NMR (400 MHz, D2O) δ 1.93-2.02 (m, 2H), 2.14-2.22 (m, 2H), 2.45-2.54 (m, 3H), 2, 63-2.68 (m, 2H), 2.88-3.56 (m, 18H), 3.57-3.72 (m, 11H), 3.73-3.85 (m, 3H), 3.86-4.10(m, 3H), 4.16(s, 1H), 4.21-4.31(m, 3H);

MS (ESI) m/z 895 [M+H] ⁺ .

Step 4 Preparation of Compound 2o.

Compound 2o (17 mg, 48%) was prepared as described for compound 2n, except that compound 23b (50 mg, 38 μmol) and 70% trifluoroacetic acid/dichloromethane (1 ml) were used.

1H NMR (400 MHz, D2O) δ 1.92-2.03 (m, 2H), 2.15-2.25 (m, 2H), 2.48-2.68 (m, 5H), 2, 71-2.75 (m, 1H), 2.92-3.53 (m, 18H), 3.55-3.86 (m, 17H), 3.87-4.17 (m, 3H), 4.22-4.31 (m, 2H);

MS (ESI) m/z 909 [M+H] ⁺ .

Example 12 Preparation of compound 2p.

Stage 1. Preparation of compound 25.

Compound 20 (500 mg, 1.02 mmol) was dissolved in dichloromethane (10 ml) and cooled to 0°C, N,N'-dicyclohexylcarbodiimide (DCC, 230 mg, 1.12 mmol) and compound 24 (170 mg , 1.02 mmol), then stirred at room temperature for 1 hour. The organic layer was filtered several times and the solvent was removed under reduced pressure. The concentrate was separated by column chromatography (30% ethyl acetate/n-hexane) to give compound 25 (450 mg, 69%).

1H NMR (400 MHz, CDQ3) δ 1.37-1.50 (m, 36H), 1.81-1.90 (m, 1H), 1.96-2.25 (m, 4H), 2, 28-2.39 (m, 3H), 2.51-2.56 (m, 1H), 4.01-4.10 (m, 2H), 4.14-4.21 (m, 2H), 4.24-4.36 (m, 3H), 5.51 (br, 1H), 5.75 (br, 1H);

- 27 041468

MS (ESI) m/z 774 [M+H]+.

Stage 2. Obtaining compound 26.

Compound 25 (350 mg, 0.55 mmol) and 2-aminoethyl 2'-azidoethyl ether (110 mg, 0.82 mmol) are dissolved in ethanol (10 ml), 1 M CuSO ₄ (0.11 ml, 0 .11 mmol) and 2 M sodium ascorbate (0.082 ml, 0.16 mmol), then stirred for 1 hour. The reagent was filtered and the solvent was removed under reduced pressure. The concentrate was separated by NH silica gel column chromatography (3% methanol/dichloromethane) to give compound 26 (250 mg, 59%).

Ή NMR (400 MHz, methanol-d ₄ ) δ 1.41-1.51 (m, 37H), 1.78-1.90 (m, 2H), 2.03-2.16 (m, 2H) , 2.29-2.42(m, 3H), 2.67-2.71(m, 1H), 3.12(q, J=5.2Hz, 2H), 3.65-3.70 (m, 2H), 3.88-3.94 (m, 2H), 4.01 (d, J=6.0 Hz, 1H), 4.16-4.22 (m, 3H), 4, 52-4.64 (m, 3H), 4.69-4.75 (m, 1H), 7.95 (s, 0.6H), 8.07 (s, 0.5H);

MS (ESI) m/z 770 [M+H] ⁺ .

Step 3: Preparation of Compound 27.

DOTA-tris(tBu) ether (36 mg, 62 µmol) was dissolved in dichloromethane (5 ml), hydroxybenzotriazole (DNA, 11 mg, 78 µmol), TBTU (25 mg, 78 µmol) and diisopropylethylamine (18 µl, 0 , 10 mmol), then stirred at room temperature for 10 min. Compound 26 (40 mg, 52 µmol) dissolved in dichloromethane (1 ml) was added thereto, followed by stirring for 1 h. The reaction was quenched by adding water (10 ml) and the organic compound was extracted using dichloromethane (10 mlx2). The reagent was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to give compound 27 (50 mg, 72%).

1H NMR (400 MHz, methanol-d4) δ 1.40-1.52 (m, 63H), 1.55-1.66 (m, 1H), 1.76-1.97 (m, 4H), 2.02-2.28 (m, 9H), 2.29-2.48 (m, 8H), 2.68-2.72 (m, 3H), 3.35-3.41 (m, 4H ), 3.48-3.71 (m, 4H), 3.81-3.87 (m, 3H), 3.94-4.08 (m, 1H), 4.13-4.23 (m , 4H), 4.52-4.59 (m, 3H), 4.64-4.75 (m, 2H), 7.92 (s, 0.6H), 8.05 (s, 0.4H );

MS (ESI) m/z 1325 [M+H] ⁺ .

Step 4 Preparation of compound 2p.

Compound 27 (43 mg, 32 µmol) was added to 70% trifluoroacetic acid/dichloromethane (1 ml), then stirred for 7 hours. After pouring the reaction mixture into diethyl ether (20 ml) to give a precipitate, it was separated using a centrifuge. The mixture was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound 2p (22 mg, 73%) as a solid.

1H NMR (400 MHz, D2O) δ 1.90-2.03 (m, 2H), 2.12-2.23 (m, 2H), 2.43-2.53 (m, 3H), 2, 72-2.78 (m, 1H), 2.99-3.46 (m, 16H), 3.52-3.59 (m, 3H), 3.62-4.10 (m, 9H), 4.14 (s, 1H), 4.19-4.35 (m, 3H), 4.57-4.63 (m, 2H), 4.65-4.74 (m, 2H), 4. 77 (s, 2H), 7.95 (s, 0.4H), 8.04 (s, 0.6H);

MS (ESI) m/z 932 [M+H] ⁺ .

Example 13 Preparation of Ga-1

Obtaining compound Ga-1a.

Compound 2a (10 mg, 11 μmol) was dissolved in water (0.6 ml), gallium trichloride (19 mg, 0.11 mmol) dissolved in water (0.6 ml) was added thereto, then stirred at 70° C. for for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1a (6 mg, 56%) as a solid.

1H NMR (400 MHz, D2O) δ 1.34-1.51 (m, 4H), 1.64-1.73 (m, 1H), 1.77-1.86 (m, 1H), 1, 91-2.00 (m, 1H), 2.12-2.20 (m, 1H), 2.50 (t, J=7.2 Hz, 2H), 3.10 (t, J=6, 8 Hz, 2H), 3.31-3.43 (m, 10H), 3.52-3.57 (m, 6H), 3.65 (s, 2H), 3.78 (s, 2H), 3.84-3.94 (m, 8H), 4.00-4.03 (m, 2H), 4.16 (dd, J=14.0, 5.2 Hz, 1H), 4.24 ( dd, J=14.0, 5.2 Hz, 1H), 4.38 (s, 2H), 4.57 (t, J=4.8 Hz, 2H), 7.88 (s, 1H);

MS (ESI) m/z 984 [M+H] ⁺ .

Obtaining compound Ga-1b.

Compound 2b (19 mg, 19 μmol) was dissolved in water (0.8 ml), gallium trichloride (33 mg, 0.19 mmol) dissolved in water (0.8 ml) was added thereto, then stirred at 70° C. for for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1b (13 mg, 64%) as a solid.

1H NMR (400 MHz, D2O) δ 1.28-1.42 (m, 3H), 1.50-1.57 (m, 2H), 1.65-1.73 (m, 1H), 1, 77-1.86 (m, 1H), 1.89-1.99 (m, 1H), 2.10-2.20 (m, 1H), 2.48 (t, J=7.2 Hz, 2H), 3.27-3.42(m, 11H), 3.49-3.57(m, 6H), 3.67(s, 2H), 3.78(s, 2H), 3.83 -3.92 (m, 8H), 4.00-4.03 (m, 2H), 4.15 (dd, J=14.0, 4.8 Hz, 1H), 4.24 ((dd, J=14.0, 4.8 Hz,

- 28 041468

1H), 4.58 (t, J=4.8 Hz, 2H), 5.19 (s, 2H), 7.62 (d, J=7.6 Hz, 2H), 8.01 (s, 1H), 8.42 (d, J=7.6 Hz, 2H);

MS (ESI) m/z 1061 [M+H]+.

Obtaining compound Ga-1c.

Compound 2c (10 mg, 10 µmol) was dissolved in water (1 ml), gallium trichloride (18 mg, 100 µmol) dissolved in water (1 ml) was added thereto, followed by stirring at 70°C for 1 hour. Reaction solution filter. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1c (6 mg, 58%) as a solid.

1H NMR (400 MHz, D ₂ O) δ 1.28-1.42 (m, 2H), 1.44-1.62 (m, 2H), 1.64-1.77 (m, 1H), 1.78-1.90 (m, 1H), 1.93-2.23 (m, 1H), 2.50-2.54 (m, 2H), 2.72-2.75 (m, 0 .5H), 2.87-2.90 (m, 1.5H), 3.01-3.07 (m, 2H), 3.36-3.44 (m, 12H), 3.54-3 .59 (m, 6H), 3.69 (s, 2H), 3.77 (s, 2H), 3.85-3.94 (m, 8H), 4.00-4.12 (m, 4H ), 4.16-4.29 (m, 3H), 4.57-4.60 (m, 2H), 7.81 (s, 1H);

MS (ESI) m/z 1041 [M+H] ⁺ .

Obtaining compound Ga-1d.

Compound 2d (6 mg, 6 µmol) was dissolved in water (0.8 ml), gallium trichloride (14 mg, 80 µmol) dissolved in water (0.8 ml) was added thereto, followed by stirring at 70° C. for 1 h. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1d (4 mg, 62%) as a solid.

1H NMR (400 MHz, D2O) δ 1.26-1.43 (m, 2H), 1.44-1.60 (m, 2H), 1.66-1.78 (m, 1H), 1, 79-1.91 (m, 1H), 1.94-1.20 (m, 1H), 2.15-2.24 (m, 1H), 2.48-2.67 (m, 4H), 2.79-2.88 (m, 2H), 3.01-3.09 (m, 2H), 3.28-3.49 (m, 11H), 3.52-3.65 (m, 8H ), 3.70 (s, 2H), 3.79 (s, 2H), 3.84-3.99 (m, 7H), 4.05 (d, J=10.8 Hz, 2H), 4 .18-4.29 (m, 2H), 4.60 (t, J=5.2 Hz, 2H), 7.82 (s, 0.6H), 7.84 (s, 0.4H);

MS (ESI) m/z 1055 [M+H] ⁺ .

Obtaining compound Ga-1e.

Compound 2e (16 mg, 18 µmol) was dissolved in distilled water (0.5 ml), gallium trichloride (16 mg, 91 µmol) was added thereto, followed by stirring at 70° C. for 1 hour. The reaction solution was filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1e (15 mg, 86%) as a white solid.

1H NMR (400 MHz, D ₂ O) δ 1.17-1.32 (m, 2H), 1.37-1.51 (m, 2H), 1.53-1.62 (m, 1H), 1.65-1.75 (m, 1H), 1.82 (n, J=7.4 Hz, 1H), 2.01 (n, J=7.6 Hz, 1H), 2.35 (t , J=7.2Hz, 2H), 3.00-3.30(m, 12H), 3.36-3.41(m, 4H), 3.47(kv, J=4.8Hz, 2H), 3.51-3.60 (m, 6H), 3.65 (s, 2H), 3.73 (s, 4H), 3.76-3.79 (m, 2H), 3.85 -3.88 (m, 2H), 3.97 (s, 2H), 4.01-4.12 (m, 3H), 4.24 (s, 2H);

MS (ESI) m/z 975 [M+H] ⁺ .

Obtaining compound Ga-1f.

Compound 2f (3.8 mg, 3.2 µmol) synthesized in Step 5 of Example 5 was dissolved in distilled water (0.5 ml), gallium trichloride (3.0 mg, 17 µmol) was added thereto, then stirred at 70 °C for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1f (2.7 mg, 68%) as a white solid.

1H NMR (400 MHz, D ₂ O) δ 1.20-1.31 (m, 4H), 1.36-1.50 (m, 5H), 1.54-1.74 (m, 5H), 1.76-1.84 (m, 2H), 1.85-1.96 (m, 1H), 1.99-2.04 (m, 2H), 2.13 (t, J=7.4 Hz, 2H), 2.35 (t, J=7.2 Hz, 2H), 2.51 (t, J=7.0 Hz, 2H), 3.03 (t, J=6.8 Hz, 2H), 3.15-3.28 (m, 9H), 3.33-3.44 (m, 8H), 3.46-3.50 (m, 3H), 3.55-3.58 ( m, 3H), 3.65-3.70 (m, 3H), 3.75-3.81 (m, 4H), 3.87 (s, 2H), 3.92-4.12 (m, 5H), 4.26 (s, 2H), 7.14-7.17 (m, 3H), 7.24-7.27 (m, 2H);

MS (ESI) m/z 1252 [M+2H]+, 1248 [M-2H] ^- .

Obtaining compound Ga-1g.

Compound 2g (4.4 mg, 3.7 µmol) synthesized in Step 5 of Example 5 was dissolved in distilled water (0.5 ml), gallium trichloride (4.0 mg, 23 µmol) was added thereto, then stirred at 70 °C for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1g (2.0 mg, 43%) as a white solid.

1H NMR (400 MHz, D2O) δ 1.12-1.28 (m, 4H), 1.32-1.37 (m, 2H), 1.40-1.48 (m, 2H), 1, 49-1.62 (m, 3H), 1.64-1.70 (m, 1H), 1.73 (p, J=7.4 Hz, 2H), 1.77-1.86 (m, 1H), 1.99-2.05 (m, 1H), 2.08 (t, J=7.2 Hz, 2H), 2.15 (s, 3H), 2.36 (t, J=7 .6 Hz, 2H), 2.43 (t, J=6.8 Hz, 2H), 2.96-3.03 (m, 2H), 3.04-3.28 (m, 1H), 3 .30-3.47 (m, 8H), 3.84-3.91 (m, 2H), 3.92-3.40 (m, 2H), 4.03-4.08 (m, 2H) , 4.09-4.14 (m, 1H), 4.23 (s, 2H), 7.01 (d, J=8.0 Hz, 2H), 7.05 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1265 [M+H] ⁺ , 1263 [MH] ^- .

Obtaining compound Ga-1h.

Compound 2h (6.0 mg, 4.6 µmol) synthesized in Step 5 of Example 5 was dissolved in distilled water (0.5 ml), gallium trichloride (6.0 mg, 34.1 µmol) was added thereto, then stirred

- 29 041468 at 70°C for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga1h (5.4 mg, 86%) as a solid.

¹ H NMR (400 MHz, D ₂ O) δ 1.21-1.30 (m, 4H), 1.33-1.38 (m, 2H), 1.40-1.51 (m, 2H) , 1.54-1.75 (m, 4H), 1.82 (n, J=7.4 Hz, 2H), 1.83-1.89 (m, 1H), 2.03-2.08 (m, 1H), 2.12 (t, J=7.2 Hz, 2H), 2.39 (t, J=7.2 Hz, 1H), 2.48 (t, J=7.2 Hz , 1H), 3.00 (t, J=6.6 Hz, 2H), 3.13-3.33 (m, 11H), 3.39-3.52 (m, 8H), 3.55- 3.60 (m, 6H), 3.66-3.74 (m, 3H), 3.97 (s, 2H), 4.00-4.17 (m, 3H), 4.27 (s, 2H), 6.94 (d, J=8.0 Hz, 2H), 7.59 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1375 [M+H]+, 1373 [MH] ^- .

Obtaining compound Ga-1i.

Compound 2i (7.0 mg, 8.1 µmol) synthesized in Step 4 of Example 6 was dissolved in distilled water (0.5 ml), gallium trichloride (7.0 mg, 40 µmol) was added thereto, then stirred at 70 °C for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1i (6.0 mg, 79%) as a white solid.

% NMR (400 MHz, D ₂ O) δ 1.22-1.33 (m, 2H), 1.40-1.52 (m, 2H), 1.56-1.64 (m, 1H), 1.68-1.76 (m, 1H), 1.85 (n, J=7.2 Hz, 1H), 2.04 (n, J=7.2 Hz, 1H), 2.38 (t , J=7.2 Hz, 2H), 3.17-3.34 (m, 11H), 3.41-3.43 (m, 4H), 3.48-3.53 (m, 2H), 3.58 (s, 4H), 3.76 (s, 4H), 3.79 (s, 2H), 3.88-3.91 (m, 2H), 3.97 (s, 2H), 4 .06-4.14 (m, 3H), 4.26 (s, 2H);

MS (ESI) m/z 933 [M+2H]+, 929 [M-2H] ^- .

Preparation of compound Ga-1j.

Compound 2j (4.4 mg, 3.8 µmol) synthesized in step 5 of Example 7 was dissolved in distilled water (0.5 ml), gallium trichloride (4.0 mg, 23 µmol) was added thereto, then stirred at 70 °C for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1j (2.3 mg, 49%) as a white solid.

% NMR (400 MHz, D ₂ O) δ 1.20-1.30 (m, 4H), 1.34-1.50 (m, 4H), 1.56-1.66 (m, 3H), 1.69-1.80 (m, 3H), 1.83-1.87 (m, 1H), 1.97-2.08 (m, 1H), 2.11 (t, J=6.8 Hz, 2H), 2.18 (s, 3H), 2.37 (t, J=7.2 Hz, 2H), 2.42-2.48 (m, 2H), 3.02 (t, J =6.4 Hz, 2H), 3.15-3.30 (m, 9H), 3.34-3.55 (m, 11H), 3.64-3.70 (m, 3H), 3, 76-3.79 (m, 4H), 3.88-3.93 (m, 4H), 4.05-4.10 (m, 3H), 4.22 (s, 2H), 7.04 ( d, J=8.0 Hz, 2H), 7.08 (d, J=8.0 Hz, 2H);

MS (ESI) m/z 1121 [M+H] ⁺ , 1119 [MH] ^- .

Obtaining compound Ga-1k.

Compound 2k (2.0 mg, 1.6 µmol) synthesized in step 5 of Example 7 was dissolved in distilled water (0.5 ml), gallium trichloride (2.0 mg, 11 µmol) was added thereto, then stirred at 70 °C for 1 hour. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1k (0.6 mg, 29%) as a white solid.

MS (ESI) m/z 1332 [M+H] ⁺ , 1330 [MH] ^- .

Obtaining compound Ga-1l.

Compound 2l (7.0 mg, 8.5 µmol) was dissolved in distilled water (0.5 ml), gallium trichloride (4.5 mg, 25.6 µmol) was added thereto, then stirred at 70°C for 2 h The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1l (5.5 mg, 73%) as a white solid.

% NMR (400 MHz, CDCl ₃ ) δ 1.24-1.40 (m, 2H), 1.42-1.51 (m, 1H), 1.54-1.62 (m, 2H), 1 .63-1.94 (m, 1H), 2.06-2.14 (m, 1H), 2.44 (t, J=7.2 Hz, 2H), 3.20-3.40 (m , 9H), 3.49 (d, J=8.8 Hz, 4H), 3.72-3.86 (m, 6H), 3.87 (d, J=10.4 Hz, 2H), 3 .92-4.00 (m, 3H), 4.04 (s, 2H), 4.10-4.20 (m, 4H);

MS (ESI) m/z 887 [M+H] ⁺ .

Obtaining compound Ga-1m.

Compound 2m (14 mg, 12.6 µmol) was dissolved in H ₂ O (0.8 ml), gallium trichloride (6.7 mg, 37.9 µmol) was added thereto, then stirred at 70°C for 2 h. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1m (7.1 mg, 48%) as a white solid.

% NMR (400 MHz, D2O) δ 1.30-1.44 (m, 4H), 1.48-1.54 (m, 2H), 1.58 (br, 2H), 1.68-1, 76 (m, 2H), 1.79-1.89 (m, 4H), 1.93-1.98 (m, 1H), 2.14-2.17 (m, 1H), 2.21 ( d, J=1.2 Hz, 4H), 2.29 (s, 3H), 2.45 (t, J=7.2 Hz, 2H), 2.54-2.57 (m, 2H), 3.15 (t, J=6.4 Hz, 2H), 2.78-3.42 (m, 9H), 3.54 (t, J=10 Hz, 4H), 3.78 (s, 3H ), 3.83-3.97 (m, 6H), 4.00-4.11 (m, 4H), 4.16-4.34 (m, 4H), 7.16 (dd, J= 17.6, 7.6 Hz, 4H).

MS (ESI) m/z 1174 [MH] ^- .

Preparation of the Ga-1n compound.

Compound 2n (9 mg, 10 µmol) was dissolved in water (1 ml), gallium trichloride (18 mg, 100 µmol) dissolved in water (1 ml) was added thereto, followed by stirring at 70°C for 1 hour. The reaction solution filter. The filtrate is separated by high performance liquid chromatography (HPLC)

- 30 041468 and dried using a lyophilizer to obtain the compound Ga-1n (4 mg, 41%) as a solid.

¹ H NMR (400 MHz, D2O) δ 1.94-2.03 (m, 2H), 2.15-2.45 (m, 2H), 2.47-2.54 (m, 3H), 2 .63-2.71 (m, 2H), 3.32-3.48 (m, 9H), 3.53-3.72 (m, 15H), 3.78 (s, 2H), 3.90 (s, 4H), 3.95 (d, J=10.4 Hz, 2H), 4.04 (d, J=11.2 Hz, 2H), 4.22 (s, 2H), 4.23 -4.33 (m, 3H);

MS (ESI) m/z 961 [M+H]+.

Obtaining compound Ga-1o.

Compound 2o (7 mg, 8 µmol) was dissolved in water (0.8 ml), gallium trichloride (14 mg, 80 µmol) dissolved in water (0.8 ml) was added thereto, followed by stirring at 70° C. for 1 h. The reaction solution is filtered. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1o (6 mg, 80%) as a solid.

¹ H NMR (400 MHz, D2O) δ 1.95-2.04 (m, 2H), 2.14-2.24 (m, 2H), 2.51-2.68 (m, 5H), 2 .73-2.76 (m, 1H), 3.37-3.48 (m, 10H), 3.54-3.79 (m, 20H), 3.91 (s, 4H), 3.96 (d, J=10.4 Hz, 2H), 4.04 (d, J=11.2 Hz, 2H), 4.23-4.30 (m, 2H);

MS (ESI) m/z 977 [M+H]+.

Obtaining compound Ga-1p.

Compound 2p (9 mg, 10 µmol) was dissolved in water (1 ml), gallium trichloride (18 mg, 10 µmol) dissolved in water (1 ml) was added thereto, followed by stirring at 70°C for 1 hour. The reaction solution filter. The filtrate was separated by high performance liquid chromatography (HPLC) and dried using a lyophilizer to give compound Ga-1p (5 mg, 51%) as a solid.

1H NMR (400 MHz, D2O) δ 1.86-2.02 (m, 2H), 2.08-2.21 (m, 2H), 2.44-2.49 (m, 3H), 2, 67-2.73 (m, 1H), 3.25-3.42 (m, 9H), 3.45-3.59 (m, 8H), 3.66 (s, 2H), 3.80- 3.95 (m, 7H), 4.00-4.10 (m, 4H), 4.15-4.19 (m, 2H), 4.56-4.59 (m, 2H), 4. 64-4.73 (m, 2H), 4.77 (s, 2H), 7.93 (s, 0.4H), 8.03 (s, 0.6H);

MS (ESI) m/z 998 [M+H]+.

Example 14 Preparation of [ ⁶⁸ Ga]1 Compound.

Obtaining compound [ ⁶⁸ Ga] 1 a.

0.1 N hydrochloric acid (5 ml) is poured into a ⁶⁸ Ge/ ⁶⁸ Ga generator and placed in test tubes (1 ml/tube). After measuring the radioactivity of each test rub, two solutions of ⁶⁸ Ga (4.6 mCi, 2 ml) from the two test tubes showing high radioactivity are transferred to the reaction vessel. Compound 2a (200 µg) was dissolved in 1.0 M sodium acetate solution (0.4 ml)-aqueous hydrochloric acid (pH 4.55), which was charged into a reaction vessel, followed by reaction at 80° C. for 10 minutes. The reaction solution was filtered, and the filtrate was separated by high performance liquid chromatography. The separated solution was diluted with water (10 ml), passed through a C-18 SepPak for absorption and washed with water (5 ml). After purging with nitrogen to remove moisture, it was eluted with ethanol (1 ml) to give the compound [ ⁶⁸ Ga]1a (1.4 cci).

High performance liquid chromatography conditions.

Column: RESTEK AQ (5 µm, 250x10 mm).

Mobile phase: 25% methanol/water (0.1% TFA).

Flow rate: 3 ml/min.

UV sensor: 230 nm.

Stay time: 12 min.

Obtaining the compound [ ⁶⁸ Ga]1b.

Compound [ ⁶⁸ Ga] 1b (2.8 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (5.94 mCi, 2 ml) and compound 2b (200 μg) were used.

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 5% ethanol/water (0.1% TFA).

Flow rate: 5 ml/min.

UV sensor: 254 nm.

Stay time: 32 min.

Obtaining the compound [ ⁶⁸ Ga]1c.

Compound [ ⁶⁸ Ga] 1c (2.5 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (5.7 mCi, 2 ml) and compound 2c (200 μg) were used.

- 31 041468

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 0-15% 30 min acetonitrile/water (0.1% TFA).

Flow rate: 3 ml/min.

UV sensor: 230 nm.

Stay time: 31 min.

Obtaining the compound [ ⁶⁸ Ga]1e.

Compound [ ⁶⁸ Ga] 1e (2.4 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (5.5 mCi, 2 ml) and compound 2e (200 μg) were used.

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 8% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 14 min.

Obtaining the compound [ ⁶⁸ Ga]1f.

Compound [ ⁶⁸ Ga]1f (1.3 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (4.6 mCi, 2 ml) and compound 2f (200 μg) were used.

High performance liquid chromatography conditions.

Column: Xterra MS C18 (10 µm, 250x10 mm).

Mobile phase: 20% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 15 min.

Obtaining the compound [ ⁶⁸ Ga]1g.

Compound [ ⁶⁸ Ga] 1g (2.3 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (5.5 mCi, 2 ml) and compound 2g (200 μg) were used.

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 25% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 17 min.

Obtaining the compound [ ⁶⁸ Ga]1h.

Compound [ ⁶⁸ Ga] 1h (1.3 mCi) is prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (4.6 mCi, 2 ml) and compound 2h (200 μg) are used.

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 30% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 14 min.

Obtaining the compound [ ⁶⁸ Ga]1k.

Compound [ ⁶⁸ Ga] 1k (1.1 mCi) is prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (3.9 mCi, 2 ml) and compound 2k (200 μg) are used.

High performance liquid chromatography conditions.

Column: Xterra MS C18 (10 µm, 250x10 mm).

Mobile phase: 25% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 18 min.

Obtaining the compound [ ⁶⁸ Ga]1n.

Compound [ ⁶⁸ Ga] 1n (3.8 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (6.9 mCi, 2 ml) and compound 2n (200 μg) were used.

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 7% acetonitrile/water (0.1% TFA).

Flow rate: 3 ml/min.

UV sensor: 220 nm.

Stay time: 13 min.

- 32 041468

Obtaining the compound [ ⁶⁸ Ga]1o.

Compound [ ⁶⁸ Ga] 1o (2.1 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (5.4 mCi, 2 ml) and compound 2o (200 μg) were used.

High performance liquid chromatography conditions.

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 7% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 20 min.

Obtaining the compound [ ⁶⁸ Ga]1p.

Compound [ ⁶⁸ Ga] 1p (2.1 mCi) was prepared according to the procedure described for the preparation of compound 1a, except that a solution ^{of 68} Ga (5.8 mCi, 2 ml) and compound 2p (200 μg) were used.

High performance liquid chromatography conditions.

Column: RESTEK AQ (250x10 mm).

Mobile phase: 15% methanol/water (0.1% TFA).

Flow rate: 3 ml/min.

UV sensor: 220 nm.

Stay time: 15 min.

Example 15 Preparation of [ ⁶⁴ Cu]1b.

An aqueous solution of hydrochloric acid in which [ ⁶⁴ Cu]CuCl ₂ (7.3 mCi) is dissolved is heated at 90° C. and dried with a nitrogen purge. After drying, 0.1 ml of 0.1 M sodium citrate (pH 5.5) in which compound 2b (100 µg) was dissolved therein was added, followed by reaction at 60° C. for 10 minutes. After completion of the reaction, water (0.3 ml) was added to the reaction mixture, filtered and washed twice with water (0.3 ml). The filtrate was separated by high performance liquid chromatography, passed through a C-18 SepPak for absorption, washed with 5 ml of water and poured out with 1 ml of ethanol to give the compound [ ⁶⁴ Cu] 1b (5.22 mCi).

High performance liquid chromatography conditions.

Column: Xterra MS C18 (10 µm, 250x10 mm).

Mobile phase: 50% acetonitrile/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 230 im.

Stay time: 17.5 min.

Example 16 Preparation of the compound [ ¹⁷⁷ Lu] 1g.

Compound 2g (200 µg) dissolved in 1.0 M sodium acetate solution (0.4 ml)-aqueous hydrochloric acid (pH 4.88) was loaded into a reaction vessel containing Lu-177 (2.2 mCi), followed by reaction at 80°C for 10 min. The reaction mixture is filtered and the filtrate is separated by high performance liquid chromatography. The separated solution was diluted with water (10 ml), passed through a C-18 SepPak for absorption and washed with water (5 ml). After purging with nitrogen to remove moisture, it was eluted with ethanol (1 ml) to give [ ¹⁷⁷ Lu] 1g (1.36 mCi).

Column: YMC-Pack ODS-A (S-5 µm, 12 nm, 250x10 mm).

Mobile phase: 25% ethanol/water (0.1% TFA).

Flow rate: 4 ml/min.

UV sensor: 220 nm.

Stay time: 19 min.

- 33 041468

Comparative Example 1 Synthesis of the compound [ ¹²⁵ I]30.

Stage 1. Preparation of compound 28.

Triphosgene (21 mg, 71 mmol) was dissolved in dichloromethane (5 ml), and 4-iodaniline (45 mg, 0.205 mmol) dissolved in dichloromethane (5 ml) at 0°C was slowly added thereto. Triethylamine (0.57 ml, 0.410 mmol) was added thereto, followed by stirring at 0° C. for 30 minutes. Compound 3 a (100 mg, 0.205 mmol) dissolved in dichloromethane (10 ml) at 0° C. was slowly added thereto, and triethylamine (0.57 ml, 0.410 mmol) was also added. The mixture is stirred for 5 hours, slowly raising the temperature to room temperature. The reaction mixture was concentrated under reduced pressure, and the concentrate was separated by column chromatography (2% methanol/dichloromethane) to give compound 28 (66 mg, 44%) as a white liquid.

1H 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=7.2Hz, 1H), 6.31 (q, J=3.4Hz, 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: Preparation of Compound 29.

Compound 28 (50 mg, 0.068 mmol) synthesized in step 1 above is dissolved in dioxane (1.0 ml), hexamethylditin ((Me ₃ Sn) ₂ , 043 ml, 0.206 mmol) and bis(triphenylphosphine)palladium dichloride are added thereto (I) ((Pd(PPh ₃ )Cl ₂ , 4.8 vu, 5 mmol) d in this order, then stirred at 110°C for 1.5 hours. The mixture was cooled to room temperature, an aqueous chloride solution was added thereto potassium (50 ml), then stirred for 1 h. The reagent is filtered and the organic compound is extracted using ethyl acetate. The collected organic solution is dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate is separated by column chromatography (triethylamine: ethyl acetate: n-hexane , 1:40:59) to give compound 29 (28 mg, 53%) as a white solid.

1H 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 (ws, 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]+.

Stage 3. Obtaining the compound [ ¹²⁵ I]28.

Compound 29 (100 mg) obtained in Step 2 above was dissolved in ethanol (0.250 ml), sodium iodide solution [ ¹²⁵ I] (3.2 mCi, 50 ml) was added thereto, followed by stirring at room temperature. 1 N hydrochloric acid aqueous solution (0.10 ml) and 3% H ₂ O ₂ were added thereto, followed by stirring at room temperature for 10 minutes. A 0.1 M sodium thiophosphate solution (0.20 ml) was added to the reaction mixture, and distilled water (18 ml) was added thereto. This solution is passed through a C-18 SepPak and washed with distilled water (20 ml). After pouring acetonitrile (2.0 ml)

- 34 041468 on C-18 Sep-Pak nitrogen is purged into the solution to remove acetonitrile.

Stage 4. Obtaining the compound [ ¹²⁵ 1]30.

Dichloromethane (0.2 ml) and trifluoroacetic acid (0.8 ml) were successively added to the reaction solution containing the reaction mixture obtained in Step 3 above, followed by stirring at room temperature for 20 minutes. The reaction solvent was removed by blowing with nitrogen, and then distilled water (2.0 ml) was added thereto. This solution was separated by high performance liquid chromatography (HPLC) to give the compound [ ¹²⁵ 1]20 (1.1 mCi, 24%).

HPLC conditions.

Column, XTerra MS C18 (250x10 mm); mobile phase, 30% acetonitrile/water (0.1% TFA); flow rate, 5 ml/min; UV, 254 mm; residence time, 10.4 min.

Reference Example 1 Obtaining colonies of prostate cancer cells and nude mice.

The colony of human prostate cancer cells (22RV1) used here is purchased from the American Type Culture Collection (ATCC). Colonies of human prostate cancer cells Rs PIP (PSMA ⁺ ) and Rs flu (PSMA) were obtained from Dr. Martin G. Pomper (Johns Hopkins Medical School, Baltimore, MD). Colonies of human prostate cancer cells are maintained in RPMI1640 medium supplemented with 10% fetal bovine serum (FCS) and 1% antibiotic/antifungal agent. Puromycin at a concentration of 2 μg/ml was additionally added to the culture of RSH PIP (PSMA ⁺ ) and RSH flu (PSMA ) cell colonies.

The test animals used are 6 week old male nude mice (Narabio, Seoul, Korea).

Experimental Example 1. Measurement of lipophilicity (logP).

Each of the compounds [ ⁶⁸ Ga]l (1~2 mCi) synthesized in example 14 is transferred into a vial and the solvent is removed, 1 ml of n-octanol and 1 ml of PRPB are added there and the lid is carefully closed, then vortexed for 1 min. After separation of the layers, 0.1 ml is taken from each layer and the radiation dose is measured. The radiation dose is measured 3 times and the average value is determined.

Table 1

LogP values of compounds [ ⁶⁸ Ga] 1

[ ⁶⁸ Ga]l logP [ ⁶⁸ Ga]lb -2.56 [ ⁶⁸ Ga]lc -2.65 [ ⁶⁸ Ga]le -2.88 [ ⁶⁸ Ga]lg -2.42 [ ⁶⁸ Ga]lh -1.89 [ ⁶⁸ Ga]ln -3.06

Experimental example 2 Measurement of binding ability.

To confirm the ability to bind compounds in accordance with this invention with PSMA conduct the following experiment.

RPMI1640 supplemented with 1% ABS (bovine serum albumin) is used as a buffer solution.

[ ¹²⁵ 1]30 (0.1 nM) obtained in Comparative Example 1 was added to a vessel containing 22RV1 cells (5 x 10 ⁴ ), the compound represented by formula 1 was loaded therein at 9 concentrations (1.00 x 10' ⁴ ~1, 00x10' ¹² M), then stirred at 37°C for 2 hours. After stirring was completed, the vessel was washed with a solution of PBS (2 ml) three times and then the radioactivity was measured using a gamma counter (2480 WIZARD2 Gamma Counter PerkinElmer Co., MA) . The 50% inhibitory concentration (1C ₅ o) for each compound was calculated using the GraphPad Prism program (GraphPad Software, Inc., CA).

In table. 2 below shows the binding affinity (1C ₅ o) of each compound and the K _d [ ¹²⁵ 1]30 value measured as 0.13 nM. (Maresca, K. R. et al., 2009, J. Med. Chern., 52, 347-357).

-35 041468

table 2

Compound _IC50 (hM) Ki value (hM) Ga la 570.70+135.97 69.88+29.78 Ga-lb 237.49+47.13 18.73+1.87 Ga-lc 21.20+2.06 3.76+0.37 Ga-ld 75.28+18.61 18.62+4.60 Ga-le 47.41+1.78 12.94+0.49 Ga-lf 25.66+5.06 8.12+1.60 Ga-lg 18.40+0.35 5.82+0.11 Ga-lh 11.00+0.35 3.00+0.10 Ga-li 68.99+3.27 14.36+0.68 Ga-lj 60.41+3.61 12.57+0.75 Ga-lk 63.85+6.09 12.70+1.07 Ga-ln 174.13+3.87 32.09+0.71 Ga-lo 1140.29+82.36 282.09+20.37 Ga-lp 702.95+144.61 129.54+26.65

As shown in Table. 2, the Ga-1a and Ga-lb compounds of Example 13 according to the present invention are compounds that do not have a carboxylic acid at the nitrogen of the lysine residue. In particular, Ga-1a shows a relatively low binding strength to PSMA. On the other hand, Ga-1c, having a structure similar to Ga-1a, is a compound having a carboxylic acid at the nitrogen in the lysine residue, and it can be seen that the PSMA binding strength is about 18.6 times higher than that of Ga-1a. This is because one (R463) of the three arginine residues, called the arginine patch, in the binding region of PSMA, and the carboxylic acid bound to the nitrogen of the lysine residue of the compound represented by formula 1 in accordance with this invention, form a strong salt bridging interaction.

In addition, the carboxylic acid of the lysine residue in the compound represented by formula 1 according to the present invention not only greatly improves the PSMA binding ability, but also increases the hydrophilicity of the compound to reduce non-specific binding in vivo and remove it more rapidly in normal organs.

The Ga-If, Ga-lg and Ga-lh compounds have a phenyl group or a substituted phenyl group, and have been confirmed to have higher bonding ability than the Ga-le compound having a similar structure without a phenyl group. Among them, the 4-iodophenyl-bound compound Ga-lh has the highest binding ability.

Experimental Example 3 MicroPET/CT imaging experiment of mice transplanted with colonies of prostate cancer cells.

A tumor model is generated by subcutaneous injection of PSMA ⁺ PC-3 PIP cells (clones of human cancer cells) into the right side of the hind paw of a nude mouse. Each of ^{the 68} Oa-labeled compounds [ ⁶⁸ Ga]le, [ ⁶⁸ Ga]lg, [ ⁶⁸ Ga]lh and [ ⁶⁸ Ga]lk is administered intravenously with 5.5-6.5 MBq (148-175 μCi/200 μl) and acquire PET/CT images using INVEON PET/CT (Siemens medical solutions, Knoxville, USA) for small animals within 60 min. At 150, 270 and 390 minutes, PET/CT images are also acquired for 30 minutes. The resulting PET/CT images are quantitatively analyzed using the Inveon™ Research Workplace (IRW).

In FIG. 1, 2, 3 and 4 are graphs showing the results of quantitative analysis of microPET/CT images for [ ⁶⁸ Ga]le, [ ⁶⁸ Ga]lg, [ ⁶⁸ Ga]lh and [ ⁶⁸ Ga]lk in % of the administered dose (ID)/ g and summarized in table. 3, 4, 5 and 6, respectively.

[ ⁶⁸ Ga]le was found to be rapidly eliminated via the kidneys and bladder at the initial stage after administration, and it was confirmed that it selectively binds to PSMA ⁺ PC-3 PIP tumors at a level of 4.05±0.64% ID/ g 270 min after injection (Fig. 1, Table 3).

In the case of [ ⁶⁸ Ga]lg, it was confirmed that the blood retention time increased due to the ability of the phenyl group to bind to albumin and tumor uptake increased with time. Compared to the [ ⁶⁸ Ga]le compound, tumor uptake of 13.00±4.95% ID/g was confirmed, which increased about 3-fold after 270 min (Fig. 2, Table 4).

In the case of [ ⁶⁸ Ga]lh and [ ⁶⁸ Ga] 1k, due to the ability of the substituted phenyl group to bind to albumin, the retention time in the blood increases, and an increase in tumor uptake over time is also confirmed, and the uptake by the tumor continues to increase after 390 min.

Compound [ ⁶⁸ Ga]lh showed tumor uptake of 16.75±0.92% ID/g at 390 min (FIG. 3, Table 5) and compound [ ⁶⁸ Ga] 1k showed tumor uptake of 18.25±4.17% ID/g after 390 min (Fig. 4, Table 6).

In addition, it was confirmed that all compounds [ ⁶⁸ Ga]le, [ ⁶⁸ Ga] 1g, [ ⁶⁸ Ga]lh and [ ⁶⁸ Ga]lk quickly

-36041468 are excreted from the body, as absorption by the kidneys decreases after 150 minutes.

Table 3

[ ⁶⁸ Ga]1e uptake by each mouse organ over time (% ID/g)

Time Tumor (pip) kidneys Bladder Liver muscles 10 2.33±0.36 21.37±9.18 19.37±0.32 4.76±0.42 1.46±0.14 20 3.64±0.49 10.96±4.32 47.31±7.34 1.96±0.22 1.38±0.11 thirty 4.08±0.60 10.88±7.09 55.82±12.46 1.29±0.05 0.95±0.13 40 4.44±0.48 14.92±13.57 60.63±15.48 1.00±0.03 0.61±0.05 50 4.56±1.03 15.84±15.60 64.24±12.76 0.80±0.02 0.53±0.05 60 4.73±0.98 15.72± 16.29 67.80±8.56 0.65±0.04 0.35±0.05 150 4.20±0.42 0.70±0.02 31.85±35.57 0.06±0.01 0.02±0.01 270 4.05±0.64 0.41±0.16 1.45±1.62 0.03±0.01 0.01±0.01

Table 4 Absorption [ ⁶⁸ Ga] 1 g each o] mouse organ over time (% ID/g) Time Tumor (pip) kidneys Bladder Liver muscles 10 2.18±0.63 16.40±0.10 2.28±0.11 9.55±2.22 1.49±1.64 20 4.42±1.50 19.80±0.73 6.98±2.51 7.26±1.06 1.88±2.00 thirty 5.43±1.94 21.18±2.17 13.51=1=5.83 6.10±0.50 1.69±1.78 40 6.43±2.32 22.74±3.48 18.89±8.84 5.38±0.22 1.61±1.69 50 7.33±2.76 24.20±4.60 22.76±10.86 4.75±0.07 1.54±1.62 60 8.29±2.99 25.78±4.72 24.74± 11.23 4.41±0.03 1.32±1.33 150 11.4±5.23 15.75=1=11.53 42.3±41.58 0.45±0.11 0.21±0.03 270 13.00±4.95 12.45±9.69 12.05±2.76 0.22±0.01 0.06±0.02

Table 5

Uptake of [ ⁶⁸ Ga]1h by each mouse organ over time (% ID/g)

Time Tumor (PIP) kidneys Uric bubble Liver muscles Heart 10 20 thirty 40 50 60 1.94±0.35 3.74±0.97 5.00±1.41 6.22±1.75 7.12±2.07 7.90±2.46 13.26±2.36 12.56±0.86 12.61±0.71 13.51±0.46 13.44±0.71 13.60±1.72 2.12±0.39 2.91±0.69 3.14±0.51 4.52±0.97 8.26±4.41 12.62±5.31 14.96±0.03 12.71±0.03 10.37±0.12 8.86±0.35 7.83±0.45 7.19±0.49 2.24±0.15 2.97±0.35 2.98±0.34 3.28±0.23 3.25±0.36 3.14±0.20 24.69±0.20 21.30±0.11 17.38±0.04 15.25±0.15 13.52±0.55 12.14±0.91 150 9.90±4.53 6.40±1.41 19.45±12.23 2.80±0.99 1.70±0.57 5.55±2.19 270 14.20±0.00 7.45±1.63 15.90±4.53 3.00±0.28 1.70±0.14 5.75±0.21 390 16.75±0.92 5.85±0.92 22.80±9.90 2.35±0.07 1.30±0.14 3.90±0.42

Table 6

Uptake of [ ⁶⁸ Ga]1k by each mouse organ over time (% ID/g)

Time Tumor (PIP) kidneys Uric bubble Liver muscles Heart 10 1.85±0.25 11.16±1.35 4.12±0.45 11.99±4.72 2.26±0.75 21.67±0.44 20 3.22±0.38 11.21±1.62 4.68±0.22 11.09±3.46 2.91±1.21 20.10±0.29 thirty 3.94±0.38 10.89±1.36 4.68±0.45 10.04±2.73 3.09±0.91 18.33±0.13 40 4.40±0.36 11.06±1.02 4.75±0.21 9.31±2.32 3.32±1.20 17.22±0.35 50 5.03±0.33 10.80±1.20 4.83±0.18 8.73±2.11 3.24±1.07 16.06±0.38 60 5.45±0.16 10.96±1.18 4.81±0.24 8.22±1.79 3.49±1.28 15.03±0.27 150 11.00±2.12 11.40±1.13 18.10±6.51 5.00±1.41 2.75±1.20 7.85±1.63 270 15.70±4.24 9.25±0.78 20.10±4.53 3.60±0.71 2.15±0.78 5.10±0.85 390 18.25±4.17 7.70±0.71 14.25±0.64 2.25±0.64 1.20±0.56 3.80±1.41

- 37 041468

Experimental Example 4 Biodistribution testing of mice transplanted with colonies of prostate cancer cells.

270 min after administration of [ ⁶⁸ Ga]1e and [ ⁶⁸ Ga]1g, microPET/CT images are taken for 30 min, then each organ is extracted (blood, muscle, fat, heart, lung, liver, spleen, stomach, intestines, kidneys). , bone and tumor) and their radioactivity is measured using a gamma counter.

In table. 7 shows uptake by each organ 5 hours after administration of [ ⁶⁸ Ga]1e or [ ⁶⁸ Ga]1g.

Biodistribution is confirmed 5 hours after compound administration. As a result, [ ⁶⁸ Ga]1g containing a phenyl group showed a higher tumor uptake rate (% VD/g) of more than 10%, which is about 1.4 times greater than the values for [ ⁶⁸ Ga]1e.

Table 7 _________Radioactivity of [ ⁶⁸ Ga]1e and [ ⁶⁸ Ga]1g in mouse organs_________

[ ⁶⁸ Ga]le [ ⁶⁸ Ga]lg Blood 0.01±0.00 0.02±0.03 muscles 0.01±0.00 0.01±0.00 Fat 0.01±0.00 0.11±0.15 Heart 0.01±0.00 0.02±0.01 Lung 0.02±0.01 0.07±0.07 Liver 0.02±0.01 0.03±0.01 Spleen 0.01±0.00 0.20±0.27 Stomach 0.02±0.02 0.02±0.02 Intestines 0.09±0.08 0.09±0.08 kidneys 0.82±0.51 9.15±11.26 Bones 0.00±0.00 0.01±0.01 PSMA+ PIP 7.34±5.49 10.48±1.05

Meanwhile, the compound represented by formula 1 according to the present invention can be formulated in various forms depending on the purpose of use. The following illustrates some formulation methods in which the compound represented by formula 1 according to the present invention is contained as an active ingredient, but the present invention is not limited thereto.

Production Example 1. Preparation of pharmaceutical formulations.

1-1. Getting powders.

Compound of formula 1 500 mg

Lactose 100 mg

Talc

Powders are obtained by mixing all of the above sized packets.

1-2. Getting pills.

Compound of Formula 1

Corn starch

Lactose is higher than mg of components that fall asleep in hermeti500

100

100 mg mg mg

magnesium stearate

Tablets are obtained by mixing all of the indicated tablet counts.

1-3. Getting capsules.

Compound of Formula 1

Corn starch

Lactose above mg components in the usual way for half 500

100

100 mg mg mg

magnesium stearate

Capsules are obtained by mixing all of the above new capsules by conventional methods to obtain capsules. 1-4. Obtaining a solution for injection.

Compound of Formula 1

Sterilized distilled water

PH regulator mg components that fall asleep in gelatin 500 mg proper amount proper amount

Solutions for injections are prepared by mixing all the above components, placing the mixture in 2 ml ampoules and sterilizing them in the usual way to obtain solutions for injections.

- 38 041468

1-5. Getting liquid formulations.

Compound of formula 1 100 mg

Isomerized sugar 10 g

Mannitol 5 g

Proper amount of purified water

All of the above components are dissolved in purified water. After adding lemon flavor, the total volume is adjusted to 100 ml by adding purified water. Liquid formulations are prepared by placing the mixture in brown bottles and sterilizing them in the usual way to obtain liquid formulations.

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

Industrial Applicability

The present invention relates to a pharmaceutical composition for the diagnosis and treatment of prostate cancer capable of targeting PSMA, and the compound provided in one aspect of the present invention comprises a glutamine-urea-lysine compound to which a radioactive metal chelator is structurally linked, and to to which an aryl group is attached, which can additionally bind to the PSMA protein. The binding between the glutamine urea-lysine compound and the chelator includes a polar spacer, which plays the role of reducing non-specific binding in vivo and exhibits a fast removal effect from the vital organs, but not from the prostate tumor. These characteristics reduce the radiation exposure caused by the radioisotope-bound therapeutic compound to normal tissues and organs and thus reduce side effects. In addition, a compound that contains a phenyl group having an albumin-binding strength has an increased blood retention time, thereby accumulating more in a prostate tumor.

Claims (12)

CLAIM 1. The compound represented by formula 1, its stereoisomer or its pharmaceutically acceptable salt Formula 1 where L1 is -(CH ₂ ) _a - where a is an integer 1~8; U is a bond or -C(O)-; R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~6; X is a bond or -C(O)-; W is a bond; L2 is a bond or -(CH ₂ )d-, where d is an integer 1~8; or Tz is a bond, L3 is C _1-12 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms; L4 is -(CH ₂ )e-, where e is an integer 1~6; n is an integer 0~1; R2 is hydrogen, C _1-5 straight or branched alkyl or halogen; Y is oxygen; Z is a chelator including a radioactive metal, where the radioactive metal is Ga-68, Cu-64 or Lu-177 and the chelator is 2. The compound, its stereoisomer or its pharmaceutically acceptable salt according to claim 1, - 39 041468 where L1 is -(CH2) _a -, where a is an integer 1~6; U is a bond or -C(O)-; R1 is -L5-CO2H, where L5 is -(CH2) _b -, where b is an integer 1~4; X is a bond or -C(O)-; W is a bond; L2 is a bond or -(CH2)d-, where d is an integer 1~6; Αν^Α Αχ^Α Ν=Ν or ^N ~ ^N i L3 is C _1-10 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms; L4 is -(CH2)e-, where e is an integer 2~4; n is an integer 0~1; R2 is hydrogen, C1.3 straight or branched alkyl or halogen; Y is oxygen; Z is a radioactive metal chelator, where the radioactive metal is Ga-68, Cu-64 and Lu-177 and is a chelator HO ₂ C—\ / \ ^CO ₂ H _N N NN' , but ₂ s—/ \__/ 3. Connection, its stereoisomer or its pharmaceutically acceptable salt according to claim 1, where L1 is -(CH ₂ ) _a -, where a is an integer 2~4; U is a bond or -C(O)-; R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~2; X is a bond or -C(O)-; W is a bond; L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~2; Tz is a bond, N=N or i L ₃ is C _1-8 straight alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms; L4 is -(CH ₂ ) ₃ -; n is an integer 0~1; R2 is hydrogen, methyl or halo; Y is oxygen; Z is a chelator including a radioactive metal, where the radioactive metal is Ga-68, Cu-64 or Lu-177 and the chelator is BUT ₂ C-x / \ ^co ₂ n NN A A g BUT ₂ C - \ __ / ^x - I 4. The compound, its stereoisomer or its pharmaceutically acceptable salt according to claim 1, where the compound represented by formula 1 is selected from the group consisting of the following compounds: - 40 041468 - 41 041468 - 42 041468 where in the above formulas M is a radioactive metal and the radioactive metal is as defined in claim 1. 5. A compound represented by formula 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof Formula 2 where L ₁ is -(CH ₂ ) _a -, where a is an integer 1~8; U is a bond or -C(O)-; R ₁ is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~6; X is a bond or -C(O)-; W is a bond; L2 is a bond or -(CH ₂ )d-, where d is an integer 1~8; or Tz is a bond, L ₃ is C _1-12 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms; L4 is -(CH ₂ )e-, where e is an integer 1~6; n is an integer 0~1; R2 is hydrogen, C _1-5 straight or branched alkyl or halogen; Y is oxygen; Z' is a chelator and a chelator is NN > HO ₂ C- ^/ 1 6. Connection, its stereoisomer or its pharmaceutically acceptable salt according to claim 5, where L ₁ is -(CH ₂ ) _a -, where a is an integer 1~6; U is a bond or -C(O)-; R ₁ is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~4; X is a bond or -C(O)-; W is a bond; L2 is a bond or -(CH ₂ ) _d - where d is an integer 1~6; or Tz is L ₃ is C _1-10 straight or branched alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms; L4 is -(CH ₂ ) _e -, where e is an integer 2~4; - 43 041468 n is an integer 0~1; R2 is hydrogen, C _1-3 straight or branched alkyl or halogen; Y is oxygen; Z' is a chelator and a chelator is HO ₂ C - \ / \ CO ₂ H N N NO ₂ S ^Y \__/ 7. Connection, its stereoisomer or its pharmaceutically acceptable salt according to claim 5, where L1 is -(CH ₂ ) _a -, where a is an integer 2~4; U is a bond or -C(O)-; R1 is -L5-CO2H, where L5 is -(CH ₂ ) _b -, where b is an integer 1~2; X is a bond or -C(O)-; W is a bond; L2 is a bond or -(CH ₂ )d-, where d is an integer 1~2; An^A Α^^νΑ Tz is a bond, N=N or ^NN ; L ₃ is C _1-8 direct alkylene, where one or more carbon atoms in the alkylene may be replaced by oxygen atoms; L ₄ is -(CH ₂ ) ₃ -; n is an integer 0~1; R2 is hydrogen, methyl or halo; Y is oxygen; Z' is a chelator and a chelator is BUT ₂ C—\ / \ /—co ₂ n ^N N HO ₂ C— ^/ \_/ V 8. A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 5, wherein the compound represented by formula 2 is selected from the group consisting of the following compounds: (3) - 44 041468 - 45 041468 - 46 041468 9. Composition for diagnosing prostate cancer, containing an effective amount of the compound, its stereoisomer or its pharmaceutically acceptable salt according to claim 1 as an active ingredient and a pharmaceutically acceptable diluent or excipient. 10. A pharmaceutical composition for the prevention or treatment of prostate cancer, containing an effective amount of a compound, stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 1 as an active ingredient and a pharmaceutically acceptable diluent or excipient. 11. The use of a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 1 for the preparation of a medicament for the diagnosis of prostate cancer. 12. The use of a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 1 for the preparation of a medicament for the treatment of prostate cancer.