KR102412174B1 - Pretargeting radiopharmaceuticals for diagnosis and therapy of cancer or inflammantory disease - Google Patents

Abstract

The compound of Formula 1 of the present invention can be diagnosed or treated through a pre-targeting method, and by binding to the albumin protein, the concentration in the blood can be maintained for a relatively long time, and the retention time can be adjusted according to the magnitude of the binding force with the albumin. It can be used for both diagnostic and therapeutic purposes.

Description

Pretargeting radiopharmaceuticals for diagnosis and therapy of cancer or inflammantory disease

The present invention relates to a pre-targeted radiopharmaceutical for diagnosis and treatment of cancer or inflammatory disease.

Due to their high specificity and binding power, antibodies have long been of interest as materials for delivering radionuclides for diagnosis and treatment of diseased tissues, and studies have been conducted on antibodies labeled with several radionuclides.

However, due to the slow distribution and poor pharmacokinetic properties of antibodies in vivo, the problem of radiation exposure has been raised from the perspective of nuclear medicine. Specifically, the antibody has a long half-life in the blood and it takes several days to reach the optimal biodistribution in the living body. In this respect, long-lived nuclides, such as Zr-89 (half-life: 3.3 days) and I-124 (half-life: 4.2 days), may remain significantly after the antibody has reached optimal biodistribution, but in normal organs for a long period of time. High radiation exposure can occur, which can cause side effects. In particular, when a therapeutic nuclide emitting high-energy beta or alpha particle radiation is used together with an antibody, it is highly likely to cause damage to normal organs due to the slow biodistribution of the antibody. Therefore, various efforts are being made to preserve the excellent properties of the antibody, but to lower the disadvantages in terms of nuclear medicine such as radiation exposure.

As part of this, low molecular weight antibody fragments with faster excretion rate have been studied bioengineering because the slow biodistribution characteristic of the antibody is due to the large molecular weight of the antibody, but radionuclide-labeled antibody fragments often show low tumor uptake. or the high intake in the kidneys did not achieve the desired purpose.

Another useful method, in vivo pretargeting, was devised in 1985. In this method, the antibody and radionuclide are used separately, and the antibody with a slow biodistribution is first injected into the human body to reach the optimal biodistribution, and then the radionuclide is injected to induce selective binding between the antibody and the radionuclide. will be.

In this regard, various methods for specifically binding two substances in vivo have been studied. Methods using the high binding affinity of biotin and Streptavidine or using the binding affinity between an antibody and radioligand introduced with complementary oligonucleotides, respectively, were also developed, and organic chemical reactions that induce irreversible covalent bonding were also developed.

Click chemistry is a reactive group capable of forming covalent bonds between specific functional groups under mild conditions and aqueous solutions, and has been applied to the binding between antibody-radioligands in vivo. Traceless Staudinger ligation using the Staudinger ligation reaction has been reported, but it has the disadvantage of low in vivo chemical stability and reaction rate [Saxon, E., JI Armstrong, and CR Bertozzi, A "traceless" Staudinger ligation for the chemoselective synthesis of amide. bonds. Org Lett, 2000. 2 (14): p. 2141-3.].

In addition, it has been reported that the 1,2,3-triazole synthesis method between an azido compound and a cyclooctyne derivative as a bio-applicable metal-free click chemistry has a significant effect for application to the pretargeting method. The use of radioligand concentrations was required [van den Bosch, SM, et al., Evaluation of strained alkynes for Cu-free click reaction in live mice. Nucl Med Biol, 2013. 40 (3): p. 415-23].

On the other hand, 1,2,4,5-Tetrazine compounds cause inverse-electron-demand Diels-Alder (IeDDA) reactions with various dienophiles, and particularly, very fast IeDDA reactions with trans-cyclooctene (TCO) compounds with high ring-strain. [Blackman, ML, M. Royzen, and JM Fox, Tetrazine ligation: fast bioconjugation based on inverse-electron-demand Diels-Alder reactivity. J Am Chem Soc, 2008. 130 (41): p. 13518-9.]. There have been many studies using this for biocompounds, and significant results have been reported in pretargeting studies. Usually, it is applied as a combination of antibody-TCO and radioisotope-Tetrazine compound.

However, the tetrazine compound to which a radioisotope is bound, which has been reported so far, has a disadvantage in that it has a high non-specific binding in vivo and a bad nuclear medicine image due to excretion through the intestine.

In addition, some radioligands without non-specific binding have a characteristic that they are excreted very quickly by the kidneys. There is this. This means that a high radiation dose is used when applied to the human body, and it is consistent with the original concept of the Pretargeting method, which makes the radioligand labeled with radioactive isotopes removed relatively quickly by compensating for the shortcomings caused by the slow biodistribution of the antibody. There is a problem in that it is difficult to diagnose the disease at an early stage and to accurately diagnose the disease at an early stage of recurrence because the sensitivity of

Accordingly, the present inventors developed a radioligand introduced with an albumin conjugate that binds to an albumin protein in the blood and applied it in vivo. As a result, the in vivo retention time of the radioligand can be adjusted according to the structure of the albumin conjugate, and this is applied to the pre-targeting method Thus, it was confirmed that it is suitable for diagnosis and treatment according to the disease, and the present invention was completed.

It is an object of the present invention to provide a diagnostic composition having excellent pharmacokinetic properties in vivo.

Another object of the present invention is to provide a compound to which a radioisotope, an albumin conjugate, and a tetrazine functional group are bound.

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

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

In order to achieve the above object,

One aspect of the present invention provides a compound comprising

1) tetrazine;

2) a ligand labeled with a radioisotope; and

3) Albumin binder.

More specifically, an aspect of the present invention provides a compound represented by the following formula (1), a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

(In Formula 1,

A, R ^a , L ¹ , L ² , L ³ , Tz, and the Albumin binder are as defined herein).

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

[Formula 2]

(In Formula 2,

A, R ^b , L ¹ , L ² , L ³ , Tz, and the Albumin binder are as defined herein).

Another aspect of the present invention provides a composition for diagnosis of cancer or inflammatory disease comprising the compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect of the present invention provides a pharmaceutical composition for the treatment of cancer or inflammation comprising the compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another aspect of the present invention, the step of injecting a compound consisting of TCO (trans-cyclooctene) bound antibody or antibody fragments (antibody fragments) (step 1); and injecting the diagnostic composition (step 2); provides a method for diagnosing cancer or inflammatory disease through pre-targeting, comprising a. Here, the antibody fragment may be a single chain fragment variable (scFv), an antigen-binding fragment (Fab), or a single-domain antibody (nanobody).

Another aspect of the present invention is cancer or inflammatory disease, comprising administering the compound represented by Formula 1, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof, to an individual or subject in need thereof treatment methods are provided.

Another aspect of the present invention provides the compound, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof for use in the treatment of cancer or inflammatory disease.

Another aspect of the present invention provides the use of the compound, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for the treatment of cancer or inflammatory disease.

The compound of Formula 1 of the present invention can bind to a pre-targeted substance in vivo through the IeDDA reaction, so it can be applied to the pre-targeting method, and since the albumin conjugate is introduced, the concentration in the blood can be maintained for a relatively long time. . In addition, since the retention time can be adjusted according to the size of the binding force with albumin, when an albumin conjugate having a low binding force with albumin is removed relatively quickly, it can be used for diagnosis. It can be maintained and used for treatment. In addition, the lipophilicity is weak, the hydrophilicity is strong, the intake in the diseased tissue is high, and the diagnosis and treatment effect are excellent.

1 is a MicroPET/CT image of a BALB/c mouse obtained for 60 minutes after administration of [ ⁶⁸ Ga] 1a .
2 is a MicroPET/CT image of a BALB/c mouse obtained for 60 minutes after administration of [ ⁶⁸ Ga] 1b .
3 is a MicroPET/CT image of a BALB/c mouse obtained for 60 minutes after administration of [ ⁶⁸ Ga] 1c .
4 is a MicroPET/CT image of a BALB/c mouse obtained for 60 minutes after administration of [ ⁶⁸ Ga] 1d .
5 is a MicroPET/CT image of a BALB/c mouse obtained for 60 minutes after administration of [ ⁶⁸ Ga] 1e .
6 is a time-radiation dose curve in the heart of BALB/c mice administered with [ ⁶⁸ Ga] 1a -[ ⁶⁸ Ga] 1e .

Hereinafter, the present invention will be described in detail.

Meanwhile, the embodiment of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiment described below. In addition, the embodiment of the present invention is provided in order to more completely explain the present invention to those of ordinary skill in the art. Furthermore, in the entire specification, "including" a certain element means that other elements may be further included, rather than excluding other elements, unless otherwise stated.

In order to provide a diagnostic composition having excellent pharmacokinetic properties in vivo, one aspect of the present invention provides a compound comprising the following.

1) tetrazine;

2) a ligand labeled with a radioisotope; and

3) Albumin binder.

More specifically, the present invention provides a compound represented by the following formula (1), a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

A is -CH- or N;

R ^a is a radioisotope-labeled ligand;

L ¹ , L ² and L ³ are independently spacers;

Tz is tetrazinyl; and

Albumin binder is an albumin binding moiety.

In Formula 1, A may be -CH-.

In Formula 1, the radioactive isotope is a positron emitting isotope, a single photon emitting isotope, an Auger electron emitting isotope, a beta particle emitting isotope, an alpha particle emitting isotope, or two or more radiations It may be an isotope of, F-18, Sc-43, Sc-44, Sc-47, Cu-62, Cu-64, Cu-67, Ga-67, Ga-68, Cr-51, Y-88 , Y-90, Ru-97, Ru-103, Tc-99m, Rh-105, Pd-109, Sn-117m, In-111, I-123, I-125, I-131, La-140, Ce -141, Pm-149, Tb-152, Tb-152, Tb-149, Tb-155, Tb-161, Sm-153, Ho-166, Dy-165, Dy-166, Tm-167, Yb-168 , Yb-175, Lu-177, Re-186, Re-188, Au-198, Au-199, Pb-203, At-211, Bi-211, Bi-212, Bi-213, Bi-214, Ra -223, Ac-225, Th-227, Th-231, and may be any one selected from the group consisting of Th-234.

In Formula 1, the spacer is absent; or amino acids, straight or branched C _1-20 alkylene, straight or branched C _{2-20 alkenylene, straight or branched C 2-20 alkynylene , -O-} , -S-. -S(=O)- -SO ₂ -, -NH-, -N=, -C(=S)-, -C(=O)- and one selected from the group consisting of C _6-10 arylene A linker consisting of a combination of the above linkers; may be.

At this time, the amino acids are alanine (Ala, A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine (Cys, C), glutamic acid (Glu, E), Glutamine (Gln, Q), Glycine (Gly, G), Histidine (His, H), Isoleucine (Ile, I), Leucine (Leu, L), Lysine (Lys, K), Methionine (Met, M), consisting of phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y) and valine (Val, V) It is one or more selected from the group, and preferably, it may be arginine (Arg, R) or glutamic acid (Glu, E).

In Formula 1, the ligand is a C _6-10 aryl, N, O, and S 5 to 10 membered heteroaryl including at least one heteroatom selected from the group consisting of, or a chelator, and the C _6- Aryl of ₁₀ may be phenyl, and heteroaryl may be pyridinyl.

In addition, the aryl and heteroaryl may be substituted with one or more substituents selected from the group consisting of -OH, straight or branched C _1-5 alkyl and straight or branched C _1-5 alkoxy, -OH, may be substituted with one or more substituents selected from the group consisting of straight-chain or branched C _1-3 alkyl and straight-chain or branched C _1-3 alkoxy, and one or more substituents selected from the group consisting of —OH and methoxy can be replaced with

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In this case, the ligand of the aryl or heteroaryl may be directly bound to a radioactive isotope, and may be labeled with a radioactive isotope, specifically, F-18, I-123, I-125 or I-131 may be bound to it. have. The ligand of the chelator may be labeled with a radioactive isotope, including a metal radioisotope.

이고, R¹은 수소, C_1-20의 지방족 탄화수소, C_6-10의 아릴, 또는 N, O 및 S로 이루어지는 군으로부터 선택되는 하나이상의 헤테로원자를 포함하는 5 내지 10원자의 헤테로아릴이고, 여기서, 지방족 탄화수소의 탄소는 질소, 산소 및 황으로 이루어진 군으로부터 선택되는 하나 이상의 헤테로 원자로 대체될 수 있고, 상기 지방족 탄화수소는 포화 또는 불포화 상태일 수 있다. 또한, 상기 R¹은 수소 또는 메틸일 수 있다.In Formula 1, tetrazinyl is

and R ¹ is hydrogen, C _1-20 aliphatic hydrocarbon, C _6-10 aryl, or 5 to 10 membered heteroaryl including at least one heteroatom selected from the group consisting of N, O and S, Here, the carbon of the aliphatic hydrocarbon may be replaced with one or more hetero atoms selected from the group consisting of nitrogen, oxygen and sulfur, and the aliphatic hydrocarbon may be in a saturated or unsaturated state. In addition, R ¹ may be hydrogen or methyl.

delete

In addition, the albumin binding moiety is C _6-10 aryl or 5 to 10 membered heteroaryl containing at least one N, and the aryl and heteroaryl are halogen, straight or branched C _1-5 alkyl, phenyl and N It may be substituted with one or more substituents selected from the group consisting of 5 or 6 membered heteroaryl containing one or more heteroatoms selected from the group consisting of , O and S.

Further, the albumin binding moiety is phenyl, naphthalenyl or pyridinyl, wherein said phenyl, naphthalenyl and pyridinyl are halogen, straight or branched chain C _1-5 alkyl, phenyl and from the group consisting of N, O and S It may be substituted with one or more substituents selected from the group consisting of 5 or 6 membered heteroaryl containing one or more selected heteroatoms.

Further, the albumin binding moiety is phenyl, naphthalenyl or pyridinyl, and the phenyl, naphthalenyl and pyridinyl may be substituted with one or more substituents selected from the group consisting of halogen, methyl and phenyl.

The compound represented by Formula 1 may be any one compound selected from the following compound group. In the following compounds, M is a metal radioactive isotope, and specifically, may be ⁶⁸ Ga.

;(One)

;

;(2)

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;(3)

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;(4)

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.(5)

.

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

[Formula 2]

In Formula 2,

A, L ¹ , L ² , L ³ , Tz, and the Albumin binder are as defined in Formula 1 above; and

R ^b is C _6-10 aryl, 5 to 10 membered heteroaryl including at least one heteroatom selected from the group consisting of N, O and S, or a chelator.

In Formula 2, R ^b is C _6-10 aryl, N, O, and S 5 to 10 membered heteroaryl including one or more heteroatoms selected from the group consisting of, or a chelator, the C _6- Aryl of ₁₀ may be phenyl, and heteroaryl may be pyridinyl.

In addition, the aryl and heteroaryl may be substituted with one or more substituents selected from the group consisting of -OH, straight or branched C _1-5 alkyl and straight or branched C _1-5 alkoxy, -OH, may be substituted with one or more substituents selected from the group consisting of straight-chain or branched C _1-3 alkyl and straight-chain or branched C _1-3 alkoxy, and one or more substituents selected from the group consisting of —OH and methoxy can be replaced with

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The compound represented by Formula 2 may be any one compound selected from the following compound group.

;(One)

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;(2)

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;(3)

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;(4)

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.(5)

.

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

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate formed by dissolving the derivative of Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid It can be prepared by filtration and drying, or by distilling the solvent and excess acid under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, a pharmaceutically acceptable metal salt can be prepared using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. The corresponding salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg silver nitrate).

The term “hydrate” refers to a compound of the present invention comprising a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolec μLar forces. or salts thereof. The hydrate of the compound represented by Formula 1 of the present invention may include a stoichiometric or non-stoichiometric amount of water that is bound by non-covalent intermolecular forces. The hydrate may contain 1 equivalent or more, preferably, 1 to 5 equivalents of water. Such a hydrate may be prepared by crystallizing the compound represented by Formula 1 of the present invention, an isomer thereof, or a pharmaceutically acceptable salt thereof from water or a solvent containing water.

The term “isomer” refers to a compound of the present invention or a salt thereof that has the same chemical formula or molecular formula but differs structurally or sterically. Such isomers include structural isomers such as tautomers, stereoisomers such as R or S isomers having an asymmetric carbon center, geometric isomers (trans, cis), and optical isomers (enantiomers). All these isomers and mixtures thereof are also included within the scope of the present invention.

The compound represented by Chemical Formula 1 or 2 according to the present invention may be prepared according to the preparation method shown in the following Examples, but this is only an example, and the present invention is not limited thereto. have.

Another aspect of the present invention provides a composition for diagnosis of cancer or inflammatory disease comprising the compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The cancer is pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, labial cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, Ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla Barter cancer, bladder cancer, peritoneal cancer ; Duodenal cancer, malignant soft tissue cancer, malignant bone cancer, lymphoma malignant, mesothelioma malignant, melanoma, eye cancer, vulvar cancer, ureter cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoma, gastrointestinal stromal cancer, Wilm Cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational chorionic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, acoustic schwannoma , pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, hematological cancer, and thymus It may be at least one selected from the group consisting of cancer.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, inflammatory bowel disease, lupus, hepatitis, cystitis, nephritis, Sjogren's syndrome (sjogren's syndrome), arteriosclerosis, multiple sclerosis, and may be at least one selected from the group consisting of acute and chronic inflammatory diseases.

In the diagnostic composition of the present invention, the albumin-binding moiety of the compound represented by Formula 1 can bind to albumin in the blood, so that the concentration in the blood can be maintained for a relatively long time, and also maintained according to the size of the binding force with albumin Since the time can be controlled, if an albumin conjugate having a low binding force to albumin is removed relatively quickly, it can be used for diagnosis. In addition, the hydrophilicity is strong, the intake in the diseased tissue is high, and the diagnostic effect is excellent.

In addition, since Tz can react with TCO (trans-cyclooctene) and IeDDA (Inverse electron demand Diels-Alder reactions), it can bind to a compound consisting of an antibody or antibody fragments to which TCO is bound. The diagnostic composition of the present invention can be used by applying a pretargeting method.

Another aspect of the present invention provides a pharmaceutical composition for the treatment of cancer or inflammation comprising the compound represented by Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The cancer is pseudomyxoma, intrahepatic biliary tract cancer, hepatoblastoma, liver cancer, thyroid cancer, colon cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, labial cancer, mycosis fungoides, acute myeloid leukemia, acute lymphoblastic leukemia, basal cell carcinoma, Ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple myeloma, gallbladder cancer, biliary tract cancer, colorectal cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, choroidal melanoma, ampulla Barter cancer, bladder cancer, peritoneal cancer ; Duodenal cancer, malignant soft tissue cancer, malignant bone cancer, lymphoma malignant, mesothelioma malignant, melanoma, eye cancer, vulvar cancer, ureter cancer, urethral cancer, cancer of unknown primary site, gastric lymphoma, gastric cancer, gastric carcinoma, gastrointestinal stromal cancer, Wilm Cancer, breast cancer, sarcoma, penile cancer, pharyngeal cancer, gestational chorionic disease, cervical cancer, endometrial cancer, uterine sarcoma, prostate cancer, metastatic bone cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoma, vaginal cancer, spinal cord cancer, acoustic schwannoma , pancreatic cancer, salivary gland cancer, Kaposi's sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, hematological cancer, and thymus It may be at least one selected from the group consisting of cancer.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, inflammatory bowel disease, lupus, hepatitis, cystitis, nephritis, Sjogren's syndrome (sjogren's syndrome), arteriosclerosis, multiple sclerosis, and may be at least one selected from the group consisting of acute and chronic inflammatory diseases.

In the pharmaceutical composition of the present invention, the albumin-binding moiety of the compound represented by Formula 1 can bind to albumin in the blood, so that the concentration in the blood can be maintained for a relatively long time. Also, depending on the size of the binding force with albumin Since the retention time can be controlled, if the albumin conjugate having a low binding force with albumin is removed relatively quickly, it can be used as a therapeutic composition. In addition, the hydrophilicity is strong, the intake in the diseased tissue is high, and the therapeutic effect is excellent.

In addition, since Tz can react with TCO (trans-cyclooctene) and IeDDA (Inverse electron demand Diels-Alder reactions), it can bind to a compound consisting of an antibody or antibody fragments to which TCO is bound. The pharmaceutical composition of the present invention can be used by applying a pretargeting method.

In the pharmaceutical composition of the present invention, the albumin-binding moiety of the compound represented by Formula 1 can bind to albumin in the blood, so that the concentration in the blood can be maintained for a relatively long time. Also, depending on the size of the binding force with albumin Since the retention time can be controlled, if it has an albumin conjugate having high binding strength with albumin, it can be maintained in the blood for a long time and used for treatment. In addition, the hydrophilicity is strong, the intake in the diseased tissue is high, and the therapeutic effect is excellent.

In addition, since Tz can react with TCO (trans-cyclooctene) and IeDDA (Inverse electron demand Diels-Alder reactions), it can bind to a compound consisting of an antibody or antibody fragments to which TCO is bound. The therapeutic pharmaceutical composition of the present invention can be used by applying a pretargeting method.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof according to the present invention may be administered in various oral and parenteral formulations during clinical administration, more preferably parenteral formulations. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include one or more compounds and at least one excipient, for example, starch, calcium carbonate, sucrose or lactose ( lactose), gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, and emulsions. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be administered in various oral and parenteral formulations during clinical administration. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations include one or more compounds and at least one excipient, for example, starch, calcium carbonate, sucrose or lactose ( lactose), gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, and emulsions. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

A pharmaceutical composition comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient may be administered parenterally, and parenteral administration is administered by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. depending on how

At this time, in order to formulate a formulation for parenteral administration, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed with water together with a stabilizer or buffer to prepare a solution or suspension, which is an ampoule or vial unit dosage form. can be manufactured with The composition may be sterilized and/or contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, salts and/or buffers for regulating osmotic pressure, and other therapeutically useful substances, and mixing, granulation, in the usual manner It can be formulated according to the method of formulation or coating.

Formulations for oral administration include, for example, tablets, pills, hard/soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, troches, etc. , dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg silica, talc, stearic acid and its magnesium or calcium salts and/or polyethylene glycol). Tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine and the like, optionally starch, agar, alginic acid or sodium salt thereof, etc. It may contain releasing or boiling mixtures and/or absorbents, colorants, flavoring agents, and sweetening agents.

A pharmaceutical composition for the prevention or treatment of cancer containing the compound represented by Formula 1, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient is administered as an individual therapeutic agent, or combined with another anticancer agent in use. It can be used in combination.

Another aspect of the present invention, the step of injecting a compound consisting of TCO (trans-cyclooctene) bound antibody or antibody fragments (antibody fragments) (step 1); and injecting the diagnostic composition (step 2); provides a method for diagnosing cancer or inflammatory disease through pre-targeting, comprising a. Here, the antibody fragment may be a single chain fragment variable (scFv), an antigen-binding fragment (Fab), or a single-domain antibody (nanobody).

Another aspect of the present invention is cancer or inflammatory disease, comprising administering the compound represented by Formula 1, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof, to an individual or subject in need thereof treatment methods are provided.

Another aspect of the present invention provides the compound, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof for use in the treatment of cancer or inflammatory disease.

Another aspect of the present invention provides the use of the compound, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for the treatment of cancer or inflammatory disease.

Hereinafter, the present invention will be described in detail through Examples and Experimental Examples to be described later.

However, the Examples and Experimental Examples to be described later only partially exemplify the present invention, and the present invention is not limited thereto.

<Example 1> Preparation of compound 8a

Step 1: Preparation of compound 5a

Fmoc-Gly-OH ( 3a , 200 mg, 0.67 mmol) was dissolved in dichloromethane (20 mL), and HOBt (130 mg, 1.00 mmol), TBTU (320 mg, 1.00 mmol), DIEA (0.23 mL, 1.35 mmol) was added. After adding, after stirring at room temperature for 10 minutes, compound 4 (88 mg, 0.67 mmol) was added and stirred for 1 hour more. After adding water (20 mL), organic compounds were extracted with dichloromethane (20 mL x 2). The organic layer was concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 5a (250 mg, 91%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.35 (t, J = 4.8 Hz, 2H), 3.48-3.54 (m, 3H), 3.58 (t, J = 4.8 Hz, 2H), 3.68 (t, J = 4.4 Hz, 2H), 3.89 (d, J = 5.6 Hz, 2H), 4.23 (t, J = 7.2 Hz, 1H), 4.43 (d, J = 7.2 Hz, 2H), 5.37 (br, 1H), 6.25 (br, 1H), 7.32 (t, J = 7.2 Hz, 2H), 7.41 (t, J = 7.6 Hz, 2H), 7.60 (d, J = 7.2 Hz, 2H), 7.70 (d, J = 7.6 Hz) , 2H); MS (ESI) m / z 410 (M+H) ⁺

Step 2: Preparation of compound 6a

Compound 5a (250 mg, 0.61 mmol) obtained in step 1 was dissolved in dichloromethane (2 mL), diethylamine (1 mL, 9.15 mmol) was added, and the mixture was stirred at room temperature for 18 hours. The organic solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 6a (97 mg, 85%).

MS (ESI) m / z 188 (M+H) ⁺

Step 3: Preparation of compound 7a

Fmoc-Gly-OH ( 3a , 150 mg, 0.52 mmol) was dissolved in dichloromethane (15 mL) and HOBt (110 mg, 0.78 mmol), TBTU (250 mg, 0.78 mmol), DIEA (0.18 mL, 1.04 mmol) were added. After stirring at room temperature for 10 minutes, the compound 6a (97 mg, 0.52 mmol) obtained in step 2 was added and stirred for an additional hour. After adding water (15 mL), organic compounds were extracted with dichloromethane (15 mL x 2). The organic solvent was concentrated under reduced pressure, and the concentrate was separated by column chromatography (2% methanol/dichloromethane) to obtain compound 7a (200 mg, 82%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 2.80 (s, 6H), 3.34 (t, J = 5.2 Hz, 2H), 3.47-3.51 (m, 2H), 3.57 (t, J = 4.8 Hz, 2H) , 3.66 (t, J = 5.6 Hz, 2H), 3.90 (d, J = 5.6 Hz, 2H), 3.95 (d, J = 5.2 Hz, 2H), 4.23 (t, J = 6.8 Hz, 1H), 4.46 (d, J = 6.8 Hz, 2H), 5.44 (br, 1H), 6.34 (br, 1H), 6.57 (br, 1H), 7.32 (dt, J = 7.6, 1.2 Hz, 2H), 7.41 (t, J = 7.2 Hz, 2H), 7.59 (d, J = 7.2 Hz, 2H), 7.77 (d, J = 8.0 Hz, 2H); MS (ESI) m / z 467 (M+H) ⁺

Step 4: Preparation of compound 8a

Compound 7a (200 mg, 0.43 mmol) obtained in step 3 was dissolved in dichloromethane (4 mL), diethylamine (0.83 mL, 6.43 mmol) was added thereto, and the mixture was stirred at room temperature for 18 hours. The organic solvent was removed under reduced pressure, the concentrate was separated using HPLC, and the purified solution was freeze-dried to obtain a solid compound 8a (65 mg, 62%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 3.37 (t, J = 4.8 Hz, 2H), 3.41 (t, J = 5.2 Hz, 2H), 3.57 (t, J = 6.0 Hz, 2H), 3.65 (t, J = 4.8 Hz, 2H), 3.75 (s, 2H), 3.93 (s, 2H); MS (ESI) m / z 245 (M+H) ⁺

<Example 2> Preparation of compound 8b

Step 1: Preparation of compound 5b

Dissolve Fmoc-Lys(Pbf)-OH ( 3b , 500 mg, 0.77 mmol) in dichloromethane (25 mL), HOBt (160 mg, 1.16 mmol), TBTU (370 mg, 1.16 mmol), DIEA (0.27 mL, 1.54) mmol) and stirred at room temperature for 10 minutes, then compound 4 (150 mg, 1.16 mmol) was added and stirred for an additional hour. After adding water (20 mL), organic compounds were extracted with dichloromethane (20 mL x 2). The organic solvent was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 5b (550 mg, 93%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.42 (s, 6H), 1.50-1.62 (m, 2H), 1.63-1.75 (m, 1H), 1.79-1.81 (m, 1H), 2.06 (s, 3H) ), 2.50 (s, 3H), 2.58 (s, 3H), 2.90 (s, 2H), 3.26-3.33 (m, 4H), 3.34-3.46 (m, 2H), 3.47-3.60 (m, 4H), 4.11-4.16 (m, 1H), 4.18-4.27 (m, 1H), 4.32 (d, J = 7.2 Hz, 2H), 6.04 (d, J = 7.6 Hz, 1H), 6.17 (br, 1H), 6.24 -6.38 (s, 2H), 7.06 (br, 1H), 7.23 (t, J = 7.2 Hz, 2H), 7.36 (t, J = 7.2 Hz, 2H), 7.54 (d, J = 7.6 Hz, 2H) , 7.72 (d, J = 7.6 Hz, 2H); MS (ESI) m / z 761 (M+H) ⁺

Step 2: Preparation of compound 6b

Compound 5b (530 mg, 0.70 mmol) obtained in step 1 was dissolved in dichloromethane (10 mL), diethylamine (1.08 mL, 10.4 mmol) was added thereto, and the mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 6b (330 mg, 88%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.40 (s, 6H), 1.51-1.79 (m, 6H), 2.09 (s, 3H), 2.50 (s, 3H), 2.57 (s, 3H), 2.95 ( s, 2H), 3.16-3.28 (m, 2H), 3.36 (t, J = 4.4 Hz, 2H), 3.37-3.48 (m, 3H), 3.56 (t, J = 5.2 Hz, 2H), 3.64 (t) , J = 4.4 Hz, 2H), 6.31 (s, 3H), 7.60-7.66 (m, 1H); MS (ESI) m / z 539 (M+H) ⁺

Step 3: Preparation of compound 7b

Fmoc-Gly-OH ( 3a , 72 mg, 0.24 mmol) was dissolved in dichloromethane (10 mL) and HOBt (50 mg, 0.36 mmol), TBTU (120 mg, 0.36 mmol), DIEA (84 μL, 0.48 mmol) was added. After adding, after stirring at room temperature for 10 minutes, compound 6b (130 mg, 0.24 mmol) was added and stirred for an additional hour. Water (10 mL) was added to terminate the reaction, and the organic compound was extracted with dichloromethane (10 mL x 2). The organic layer was concentrated under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 7b (190 mg, 96%).

MS (ESI) m / z 819 (M+H) ⁺

Step 4: Preparation of compound 8b

Compound 7b (190 mg, 0.23 mmol) obtained in step 3 was dissolved in dichloromethane (4 mL), diethylamine (0.36 mL, 3.48 mmol) was added thereto, and the mixture was stirred at room temperature for 18 hours. The organic solvent was removed under reduced pressure, the concentrate was separated using HPLC, and the purified solution was freeze-dried to obtain a solid compound 8b (0.13 g, 94%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.46 (s, 6H), 1.54-1.72 (m, 3H), 1.77-1.84 (m, 1H), 2.09 (s, 3H), 2.51 (s, 3H) ), 2.58 (s, 3H), 3.01 (s, 2H), 3.13-3.23 (m, 2H), 3.24-3.26 (m, 1H), 3.33-3.38 (m, 2H), 3.39-3.46 (m, 1H) ), 3.56 (t, J = 5.2 Hz, 2H), 3.63 (t, J = 5.2 Hz, 2H), 3.73 (d, J = 2.8 Hz, 2H), 4.42 (dd, J = 8.4, 5.2 Hz, 1H) ); MS (ESI) m / z 956 (M+H) ⁺

<Example 3> Preparation of compound 8c

Step 1: Preparation of compound 7c

Compound 3c (132 mg, 0.25 mmol) was dissolved in dichloromethane (20 mL), and HOBt (50 mg, 0.375 mmol), TBTU (120 mg, 0.375 mmol), DIEA (131 μL, 0.75 mmol) were added in this order, and then at room temperature was stirred for 10 minutes. Compound 6a obtained in step 2 of Example 1 was diluted in dichloromethane (5 mL), slowly added dropwise to the reaction solution, and stirred at room temperature for 30 minutes. After removing the solvent under reduced pressure, the concentrate was subjected to column chromatography (2% methanol/dichloromethane) to obtain compound 7c (171 mg, 98%).

MS (ESI) m/z 720 [M+Na] ⁺ .

Step 2: Preparation of compound 8c

Compound 7c (115 mg, 0.165 mmol) obtained in step 1 was dissolved in dichloromethane (50 mL), and diethylamine (256 μL, 2.475 mmol) was added. After stirring for 12 hours, the mixture was concentrated under reduced pressure and then column chromatography (5% methanol/dichloromethane) was performed to obtain compound 8c (74 mg, 94%) in the form of a white powder.

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.37-1.46 (m, 2H), 1.47-1.54 (m, 2H), 1.54-1.62 (m, 1H), 1.79-1.86 (m, 1H), 1.94 (p , J = 7.6, 2H), 2.16 (t, J = 7.6, 2H), 2.31 (s, 3H), 2.60 (t, J = 7.6, 2H), 3.24 (q, J = 6.4, 2H), 3.37 ( t, J = 4.8, 2H), 3.41 (dd, J = 4.6, 7.8 1H), 3.45-3.50 (m, 2H), 3.56 (t, J = 5.0 2H), 3.66 (t, J =5.0, 2H) , 3.92 (dd, J = 3.6, 5.6, 2H), 5.73 (t, J = 5.4, 1H), 6.45 (t, J =5.2, 1H), 7.05-7.10 (m, 4H), 7.88 (t, J = 5.6, 1H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 21.0, 22.8, 27.3, 29.3, 34.3, 34.8, 35.9, 38.9, 39.2, 42.8, 50.6, 54.8, 69.6, 70.1, 128.4, 129.1, 135.4, 138.4, 169.1, 172.9 , 175.7; MS (ESI) m/z 476 [M+H] ⁺ .

<Example 4> Preparation of compound 8d

Step 1: Preparation of compound 5c

Fmoc-Glu(OtBu)-OH ( 3d , 300 mg, 0.71 mmol) was dissolved in dichloromethane (20 mL), HOBt (140 mg, 1.06 mmol), TBTU (340 mg, 1.06 mmol), and DIEA (0.25 mL, 1.41 mmol) were added, followed by stirring at room temperature for 10 minutes. Then, compound 4 (40 mg, 1.06 mmol) was added and stirred for 1 hour more. Water (20 mL) was added, and organic compounds were extracted with dichloromethane (20 mL x 2). The organic layer was concentrated under reduced pressure, and the concentrate was separated by column chromatography (60% ethyl acetate/ n -hexane) to obtain compound 5c (350 mg, 92%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.46 (s, 9H), 1.88-2.02 (m, 1H), 2.04-2.16 (m, 1H), 2.24-2.36 (m, 1H), 2.37-2.46 (m, 1H), 3.36 (t, J = 4.4 Hz, 2H), 3.46-3.53 (m, 2H), 3.56-3.60 (m, 2H), 3.65 (t, J = 4.8 Hz, 2H), 4.21 ( t, J = 6.8 Hz, 2H), 4.38 (d, J = 7.2 Hz, 2H), 5.70 (d, J = 6.8 Hz, 1H), 6.53 (br, 1H) 7.32 (t, J = 7.2 Hz, 2H) ), 7.40 (t, J = 7.2 Hz, 7.60 (d, J = 7.2 Hz, 2H), 7.76 (d, J = 7.6 Hz, 2H); MS (ESI) m / z 538 (M+H) ⁺

Step 2: Preparation of compound 6c

5c (350 mg, 0.65 mmol) obtained in step 1 was dissolved in dichloromethane (6 mL) and diethylamine (1 mL, 9.77 mmol) was added thereto, followed by stirring at room temperature for 18 hours. The organic solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 6c (190 mg, 93%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.45 (s, 9H), 1.75-1.84 (m, 1H), 1.87-1.96 (m, 1H), 2.31 (t, J = 8.0 Hz, 2H), 3.32-3.48 (m, 5H), 3.58 (t, J = 5.6 Hz, 2H), 3.66 (t, J = 4.8 Hz, 2H); MS (ESI) m / z 316 (M+H) ⁺

Step 3: Preparation of compound 7d

Compound 3c (130 mg, 0.25 mmol) was dissolved in dichloromethane (10 mL), HOBt (50 mg, 0.37 mmol), TBTU (120 mg, 0.37 mmol), DIEA (0.086 mL, 0.49 mmol) were added, and then at room temperature After stirring for 10 minutes, compound 6c (0.078 g, 0.25 mmol) obtained in step 2 was added and stirred for an additional hour. Water (10 mL) was added, and organic compounds were extracted with dichloromethane (10 mL x 2). The organic layer was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 7d (180 mg, 88%).

MS (ESI) m / z 827 (M+H) ⁺

Step 4: Preparation of compound 8d

Compound 7d (180 mg, 0.22 mmol) obtained in step 3 was dissolved in dichloromethane (2 mL) and diethylamine (0.34 mL, 3.27 mmol) was added thereto, followed by stirring at room temperature for 18 hours. The organic solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 8d (100 mg, 76%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.34-1.42 (m, 2H), 1.45 (s, 9H), 1.47-1.62 (m, 4H), 1.64-1.74 (m, 1H), 1.83-1.94 (m, 3H), 2.00-2.09 (m, 1H), 2.18 (t, J = 6.8 Hz, 2H), 2.28 (s, 3H), 2.30-2.34 (m, 2H), 2.57 (t, J = 7.6) Hz, 2H), 3.17 (t, J = 7.2 Hz, 2H), 3.33-3.40 (m, 3H), 3.41-3.47 (m, 2H), 3.54-3.57 (m, 2H), 3.62-3.65 (m, 2H), 4.34 (dd, J = 8.4, 5.6 Hz, 1H), 7.04-7.09 (m, 4H); MS (ESI) m / z 604 (M+H) ⁺

<Example 5> Preparation of compound 8e

Step 1: Preparation of compound 7e

Compound 3c (130 mg, 0.24 mmol) was dissolved in dichloromethane (10 mL) and HOBt (49 mg, 0.36 mmol), TBTU (120 mg, 0.36 mmol), DIEA (0.084 mL, 0.48 mmol) were added thereto, and then at room temperature After stirring for 10 minutes, compound 6b (130 mg, 0.24 mmol) obtained in Example 2 was added and stirred for an additional hour. Water (10 mL) was added, and organic compounds were extracted with dichloromethane (10 mL x 2). The organic layer was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 7e (230 mg, 91%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.34-1.38 (m, 4H), 1.41 (s, 6H), 1.47-1.58 (m, 4H), 1.61-1.72 (m, 2H), 1.73-1.82 (m, 2H), 1.83-1.90 (m, 2H), 2.05 (s, 3H), 2.17 (t, J = 7.6 Hz, 2H), 2.24 (s, 3H), 2.50 (s, 3H), 2.53 ( d, J = 8.0 Hz, 1H), 2.58 (s, 3H), 2.81 (s, 1H), 2.94 (s, 2H), 3.08-3.18 (m, 2H), 3.32-3.41 (m, 2H), 3.53 (t, J = 5.2 Hz, 2H), 3.57 (t, J = 5.2 Hz, 2H), 3.69-3.75 (m, 2H), 4.02-4.08 (m, 1H), 4.16 (t, J = 6.8 Hz, 1H), 4.32-4.36 (m, 3H), 6.99-7.40 (m, 4H), 7.29 (dt, J = 7.6, 1.2 Hz, 2H), 7.37 (t, J = 7.6 Hz, 2H), 7.64 (d) , J = 7.6 Hz, 2H), 7.78 (d, J = 7.2 Hz, 2H); MS (ESI) m / z 1050 (M+H) ⁺

Step 2: Preparation of compound 8e

Compound 7e (230 mg, 0.22 mmol) obtained in step 1 was dissolved in dichloromethane (10 mL), and diethylamine (0.34 mL, 3.29 mmol) was added thereto. After stirring at room temperature for 18 hours, the organic solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 8e (150 mg, 83%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.29 (t, J = 7.6 Hz, 2H), 1.35-1.42 (m, 2H), 1.45 (s, 6H), 1.47-1.82 (m, 8H), 1.83-1.91 (m, 2H), 2.08 (s, 3H), 2.18 (t, J = 6.8 Hz, 2H), 2.21 (s, 3H), 2.51 (s, 3H), 2.55 (d, J = 7.6 Hz) , 1H), 2.58 (s, 3H), 2.99 (s, 2H), 3.16 (t, J = 7.2 Hz, 3H), 3.35 (t, J = 5.2 Hz, 2H), 3.38-3.45 (m, 2H) , 3.53 (t, J = 5.6 Hz, 2H), 3.61 (t, J = 5.2 Hz, 2H), 4.36 (dd, J = 8.8, 5.6 Hz, 1H), 7.03-7.07 (m, 4H); MS (ESI) m / z 828 (M+H) ⁺

<Preparation Example 1> Preparation of compound 13

Step 1: Preparation of compound 10

Compound 9 (3.0 g, 19.21 mmol) was put in a pressure tube and dissolved in acetonitrile (9.7 mL, 185.38 mmol) (Nickel(II) trifluoromethanesulfonate (3.4 g, 9.605 mmol) and hydrazine hydrate (45 mL, 922.08 mmol) were added. Nitrogen gas was filled and stirred for 12 hours at 60 ^o C. After the temperature was lowered, tetrahydrofuran (60 mL) and acetic acid (140 mL) were slowly added. Sodium nitrite (26.5 g 384 mmol) was added to water. Dissolve in 100 mL to prepare an aqueous solution, and then slowly inject it using a dropping funnel.After stirring for 15 minutes, add water, extract organic compounds using ethyl acetate, and remove moisture with anhydrous sodium sulfate.Concentrate under reduced pressure, then column chromatography (2% ethyl acetate/dichloromethane) to obtain a red solid compound 10 (750 mg, 17%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 1.47 (s, 9H), 3.08 (s, 3H), 4.96 (s, 2H), 5.65 (br s, 1H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 21.1,28.3,43.4,80.3,166.4,168.4; MS (ESI) m/z 248 [M+Na] ⁺

Step 2: Preparation of compound 11

Compound 10 (64 mg, 0.284 mmol) obtained in step 1 was dissolved in 4.0 M HCl solution (in dioxane, 15 mL) and stirred at room temperature for 1 hour. The reaction solution was precipitated in diethyl ether, centrifuged and dried to obtain a red solid compound 11 (36 mg, 78%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 3.06 (s, 3H), 4.83 (s, 2H)

Step 3: Preparation of compound 12

Compound 11 (100 mg, 0.62 mmol) obtained in step 2 was dissolved in methanol (10 mL), succinic anhydride (74 mg, 0.74 mmol) and triethylamine (0.26 mL, 1.86 mmol) were added, followed by stirring for 2 hours. . The solvent was removed under reduced pressure, and the mixture was separated by column chromatography on silica (COOH) (5% methanol/dichloromethane) to obtain compound 12 (108 mg, 77%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.31 (t, J =7.6, 2H), 2.60 (s, 3H), 3.00 (s, 2H), 3.21 (q, J =7.5, 2H)

Step 4: Preparation of compound 13

Compound 12 (80 mg, 0.62 mmol) obtained in the above step was dissolved in dichloromethane (20 mL), EDCI (204 mg, 1.065 mmol) and hydroxyl succinimide (82 mg, 0.71 mmol) were added thereto, followed by stirring for 1 hour. Water was added, organic compounds were extracted using dichloromethane, and moisture was removed with anhydrous sodium sulfate. The solvent was removed under reduced pressure and the mixture was separated by column chromatography (3% methanol/dichloromethane) to obtain a red solid compound 13 (27 mg, 24%).

MS (ESI) m/z 345 [M+Na] ⁺

<Example 6> Preparation of compound 2a

Step 1: Preparation of compound 15a

Compound 14a (72 mg, 0.13 mmol) was dissolved in dichloromethane (4 mL), and HOBt (22 mg, 0.17 mmol), TBTU (53 mg, 0.17 mmol), DIEA (38 μL, 0.22 mmol) were added in sequence. , After stirring at room temperature for 10 minutes, compound 8a (27 mg, 0.11 mmol) obtained in Example 1 was added and stirred for an additional hour. The organic solvent was removed under reduced pressure, the concentrate was separated using HPLC, and the purified solution was freeze-dried to obtain a solid compound 15a (60 mg, 71%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.44-1.56 (m, 27H), 1.93-2.02 (m, 1H), 2.09-2.18 (m, 1H), 2.45 (t, J = 7.2 Hz, 2H) ), 2.72-2.95 (m, 2H), 2.96-3.16 (m, 6H), 3.17-3.29 (m, 4H), 3.37 (t, J = 4.8 Hz, 2H), 3.57 (t, J = 6.0 Hz, 3H), 3.65 (t, J = 4.8 Hz, 3H), 3.68-3.78 (m, 1H), 3.86-3.88 (m, 3H), 3.99 (br, 2H); MS (ESI) m / z 770 (M+H) ⁺

Step 2: Preparation of compound 16a

Compound 15a (60 mg, 0.078 mmol) obtained in step 1 was dissolved in MeOH (3 mL), Palladium on carbon (10 wt. % loading, 4 mg, 0.004 mmol) was added, and hydrogen gas was filled in the reaction vessel and at room temperature Stirred for 1 hour. The reaction product was filtered through celite, the solvent was removed under reduced pressure, and the concentrate was separated by silica (NH) column chromatography (5% methanol/dichloromethane) to obtain compound 16a (38 mg, 65%).

MS (ESI) m / z 745 (M+H) ⁺

Step 3: Preparation of compound 17a

Compound 16a (38 mg, 0.051 mmol) obtained in step 2 was added to 70% trifluoroacetic acid/dichloromethane (1 mL) and stirred for 3 hours. After precipitation by adding diethyl ether (20 mL), it was separated using a centrifuge. The precipitated compound was separated by HPLC, and the purified solution was lyophilized to obtain a solid compound 17a (27 mg, 92%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.99-2.09 (m, 1H), 2.14-2.24 (m, 1H), 2.51 (t, J = 7.6 Hz, 2H), 2.78-3.29 (m, 14H) ), 3.43 (t, J = 5.6 Hz, 2H), 3.58 (t, J = 5.6 Hz, 2H), 3.64 (t, J = 10.4 Hz, 1H), 3.69 (t, J = 5.2 Hz, 2H), 3.70-3.92 (m, 6H), 3.93-4.14 (m, 2H); MS (ESI) m / z 576 (M+H) ⁺

Step 4: Preparation of compound 2a

Compound 17a (10 mg, 0.017 mmol) obtained in step 3 was dissolved in DMF (0.2 mL), DIEA (9 μL, 0.052 mmol) was added, and compound 13 obtained in Preparation Example 1 (9 mg, 0.026 mmol) was dissolved in DMF (0.3 mL), added to the reaction solution, and stirred at room temperature for 2 hours. The reaction was filtered, diluted with water, separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 2a (5 mg, 37%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.90-2.07 (m, 1H), 2.12-2.23 (m, 1H), 2.49-2.55 (m, 4H), 2.61-2.64 (m, 2H), 2.50 -2.98 (m, 3H), 3.00 (s, 3H), 3.23-3.18 (m, 5H), 3.20-3.29 (m, 3H), 3.33-3.37 (m, 2H), 3.47-3.54 (m, 4H) , 3.62-3.66 (m, 1H), 3.67-3.77 (m, 1H), 3.78-3.83 (m, 1H), 3.84-3.90 (m, 4H), 3.92-4.05 (m, 2H), 4.94 (s, 2H); MS (ESI) m / z 783 (M+H) ⁺

<Example 7> Preparation of compound 2b

Step 1: Preparation of compound 15b

Compound 14a (44 mg, 0.08 mmol) was dissolved in dichloromethane (5 mL), HOBt (14 mg, 0.10 mmol), TBTU (32 mg, 0.10 mmol), DIEA (23 μL, 0.13 mmol) were added, and then at room temperature After stirring for 10 minutes, compound 8b (40 mg, 0.067 mmol) obtained in Example 2 was added, and the mixture was further stirred for 1 hour. The organic solvent was removed under reduced pressure, the concentrate was separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 15b (75 mg, 99%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.44-1.56 (m, 27H), 1.93-2.02 (m, 1H), 2.09-2.18 (m, 1H), 2.45 (t, J = 7.2 Hz, 2H) ), 2.72-2.95 (m, 2H), 2.96-3.16 (m, 6H), 3.17-3.29 (m, 4H), 3.37 (t, J = 4.8 Hz, 2H), 3.41 (t, J = 6.0 Hz, 2H), 3.57 (t, J = 6.0 Hz, 3H), 3.65 (t, J = 4.8 Hz, 3H), 3.68-3.78 (m, 1H), 3.86-3.88 (m, 3H), 3.99 (br, 2H) ); MS (ESI) m / z 1122 (M+H) ⁺

Step 2: Preparation of compound 16b

Compound 15b (75 mg, 0.067 mmol) obtained in step 1 was dissolved in MeOH (3 mL), Palladium on carbon (10 wt. % loading, 4 mg, 0.003 mmol) was added, followed by filling the reaction vessel with hydrogen gas and at room temperature Stirred for 1 hour. The reaction product was filtered through celite, the solvent was removed under reduced pressure, the concentrate was separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 16b (66 mg, 90%).

MS (ESI) m / z 1096 (M+H) ⁺

Step 3: Preparation of compound 17b

Compound 16b (66 mg, 0.060 mmol) obtained in step 2 was dissolved in TFA:TIS:H ₂ O (95:2.5:2.5) solution (2.5 mL) and stirred at room temperature for 3 hours. Diethyl ether (20 mL) was added for precipitation, and the mixture was separated using a centrifuge. The crude compound was separated by HPLC, and the purified solution was lyophilized to obtain a solid compound 17b (22 mg, 54%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.56-1.78 (m, 3H), 1.84-1.96 (m, 1H), 1.98-2.08 (m, 1H), 2.12-2.24 (m, 1H), 2.51 (t, J = 6.8 Hz, 2H), 2.72-3.28 (m, 16H), 3.35-3.49 (m, 2H), 3.56-3.64 (m, 3H), 3.67 (t, J = 5.2 Hz, 2H), 3.72-3.90 (m, 4H), 3.91-4.10 (m, 2H), 4.32 (dd, J = 8.8, 5.6 Hz, 1H); MS (ESI) m / z 675 (M+H) ⁺

Step 4: Preparation of compound 2b

Compound 17b (10 mg, 0.015 mmol) obtained in step 3 was dissolved in DMF (0.4 mL), DIEA (8 μL, 0.044 mmol) was added, and compound 13 obtained in Preparation Example 1 (7 mg, 0.022 mmol) was dissolved in DMF (0.3 mL), added to the reaction solution, and stirred at room temperature for 2 hours. The reaction was filtered, diluted with water, separated by HPLC, and the purified solution was freeze-dried to obtain a purple solid compound 2b (6 mg, 46%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.56-1.76 (m, 3H), 1.84-1.94 (m, 1H), 1.96-2.06 (m, 1H), 2.14-2.25 (m, 1H), 2.49 -2.55 (m, 3H), 2.62-2.65 (m, 2H), 2.70-2.98 (m, 4H), 3.01 (s, 3H), 3.02-3.10 (m, 2H), 3.12-3.28 (m, 7H) , 3.33-3.46 (m, 4H), 3.48-3.54 (m, 7H), 3.58-3.61 (m, 1H), 3.62-3.76 (m, 1H), 3.78-3.99 (m, 4H), 4.38 (dd, J = 8.0, 5.6 Hz, 1H), 4.94 (s, 2H); MS (ESI) m / z 883 (M+H) ⁺

<Example 8> Preparation of compound 2c

Step 1: Preparation of compound 15c

Compound 14a (30 mg, 0.055 mmol) was dissolved in dichloromethane (10 mL), and HOBt (11 mg, 0.083 mmol), TBTU (27 mg, 0.083 mmol), DIEA (29 μL, 0.165 mmol) were added in this order, and then at room temperature stirred in. Compound 8c (31 mg, 0.066 mmol) obtained in Example 3 was dissolved in dichloromethane (5 mL), and then slowly added to the reaction solution and stirred at room temperature for 1 hour. The solvent was removed under reduced pressure, separated by HPLC, and the purified solution was freeze-dried to obtain compound 15c (24 mg, 43%) as a white solid.

MS (ESI) 1002 m/z [M+H] ⁺ ;

Step 2: Preparation of compound 16c

Compound 15c (24 mg, 0.024 mmol) obtained in step 1 was dissolved in methanol (5 mL), Palladium on carbon (10 wt. % loading, 1.3 mg, 0.0012 mmol) was added, and then the reaction vessel was filled with hydrogen gas. It was stirred at room temperature for 1 hour. The reaction solution was filtered through Celite, the solvent was removed under reduced pressure, and separated by HPLC. The purified solution was freeze-dried to obtain a white solid compound 16c (20 mg, 86%).

MS (ESI) m/z 976 [M+H] ⁺ .

Step 3: Preparation of compound 17c

Compound 16c (20 mg, 0.02 mmol) obtained in step 2 was dissolved in 60% trifluoroacetic acid/dichloromethane (1 mL), stirred for 4 hours, and then precipitated in diethyl ester. After centrifugation, the mixture was separated by HPLC, and the purified solution was freeze-dried to obtain a white solid compound 17c (12 mg, 73%).

MS (ESI) m/z 808 [M+H] ⁺

Step 4: Preparation of compound 2c

Compound 17c (12 mg, 0.015 mmol) obtained in step 3 was dissolved in DMF (0.5 mL), and compound 13 (7 mg, 0.022 mmol) obtained in Preparation Example 1 was added. DIEA (8 μL, 0.045 mmol) was added to the reaction solution and reacted at room temperature for 30 minutes. After removal of the solvent under reduced pressure and separation by HPLC, the purified solution was freeze-dried to obtain a purple solid compound 2c (8.4 mg, 56%).

MS (ESI) m/z 1013 [MH] ^-

<Example 9> Preparation of compound 2d

Step 1: Preparation of compound 15d

Compound 14a (43 mg, 0.08 mmol) was dissolved in dichloromethane (5 mL), and HOBt (14 mg, 0.10 mmol), TBTU (32 mg, 0.10 mmol), DIEA (23 μL, 0.13 mmol) were added in sequence. , After stirring at room temperature for 10 minutes, compound 8d (0.040 g, 0.067 mmol) obtained in Example 4 was added and stirred for an additional hour. The organic solvent was removed under reduced pressure, and the concentrate was separated by column chromatography (5% methanol/dichloromethane) to obtain compound 15d (66 mg, 88%).

MS (ESI) m / z 1130 (M+H) ⁺

Step 2: Preparation of compound 16d

Compound 15d (66 mg, 0.058 mmol) obtained in step 1 was dissolved in MeOH (2 mL), Palladium on carbon (10 wt. % loading, 3 mg, 0.003 mmol) was added, and then filled with hydrogen gas in a reaction vessel and then at room temperature stirred for 1 hour. The reaction product was filtered through celite, the solvent was removed under reduced pressure, the concentrate was separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 16d (40 mg, 62%).

MS (ESI) m / z 1104 (M+H) ⁺

Step 3: Preparation of compound 1 7d

Compound 16d (40 mg, 0.036 mmol) obtained in step 2 was added to 70% trifluoroacetic acid/dichloromethane (2 mL) and stirred for 3 hours. Diethyl ether (20 mL) was added for precipitation, and the mixture was separated using a centrifuge. The crude compound was separated by HPLC and the purified solution was freeze-dried to obtain a solid compound 17d (16 mg, 50%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.33-1.62 (m, 4H), 1.64-2.24 (m, 10H), 2.28 (s, 3H), 2.38-2.44 (m, 2H), 2.45-2.54 (m, 2H), 2.57 (t, J = 7.6 Hz, 2H), 2.70-3.28 (m, 16H), 3.32-3.49 (m, 2H), 3.55-3.60 (m, 3H), 3.64-3.90 (m) , 4H), 3.98 (br, 2H), 4.22 (dd, J = 8.4, 5.6 Hz, 1H), 4.30 (dd, J = 8.4, 5.6 Hz, 1H), 7.04-7.09 (m, 4H); MS (ESI) m / z 880 (M+H) ⁺

Step 4: Preparation of compound 2d

Compound 17d (12 mg, 0.014 mmol) obtained in step 3 was dissolved in DMF (0.4 mL), DIEA (7 μL, 0.041 mmol) was added, and compound 13 obtained in Preparation Example 1 (7 mg, 0.02 mmol) was added It was dissolved in DMF (0.3 mL), added to the reaction solution, and stirred at room temperature for 2 hours. The reaction solution was filtered, separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 2d (11 mg, 74%).

¹ H NMR (400 MHz, D ₂ O) δ 1.33-1.39 (m, 2H), 1.46-1.52 (m, 2H), 1.69-1.80 (m, 2H), 1.83-1.91 (m, 2H), 1.92- 2.00 (m, 2H), 2.05-2.12 (m, 2H), 2.21-2.24 (m, 2H), 2.27 (s, 3H), 2.37-2.48 (m, 4H), 2.54-2.57 (m, 4H), 2.64-2.68 (m, 2H), 2.96-3.08 (m, 6H), 3.12-3.18 (m, 5H), 3.19-3.26 (m, 4H), 3.30-3.43 (m, 4H), 3.50 (t, J ) = 7.2 Hz, 1H), 3.55-3.58 (m, 4H), 3.80-3.89 (m, 4H), 4.24 (dd, J = 8.4, 5.6 Hz, 1H), 4.31 (dd, J = 8.4, 5.6 Hz, 1H), 4.97 (s, 2H), 7.14 (dd, J = 13.2, 8.4 Hz, 4H); MS (ESI) m / z 1087 (M+H) ⁺

<Example 10> Preparation of compound 2e

Step 1: Preparation of compound 15e

Compound 14a (130 mg, 0.24 mmol) was dissolved in dichloromethane (10 mL), HOBt (49 mg, 0.36 mmol), TBTU (120 mg, 0.36 mmol), DIEA (0.084 mL, 0.48 mmol) were added, and then at room temperature After stirring for 10 minutes, compound 8e (130 mg, 0.24 mmol) obtained in Example 5 was added and stirred for an additional hour. After adding water (10 mL), the organic compound was extracted with dichloromethane (10 mL x 2), the organic layer was concentrated under reduced pressure, and the concentrate was separated by column chromatography (3% methanol/dichloromethane) to obtain compound 15e (230 mg). , 91%) was obtained.

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.34-1.38 (m, 4H), 1.41 (s, 6H), 1.47-1.58 (m, 4H), 1.61-1.72 (m, 2H), 1.73-1.82 (m, 2H), 1.83-1.90 (m, 2H), 2.05 (s, 3H), 2.17 (t, J = 7.6 Hz, 2H), 2.24 (s, 3H), 2.50 (s, 3H), 2.53 ( d, J = 8.0 Hz, 1H), 2.58 (s, 3H), 2.81 (s, 1H), 2.94 (s, 2H), 3.08-3.18 (m, 2H), 3.32-3.41 (m, 2H), 3.53 (t, J = 5.2 Hz, 2H), 3.57 (t, J = 5.2 Hz, 2H), 3.69-3.75 (m, 2H), 4.02-4.08 (m, 1H), 4.16 (t, J = 6.8 Hz, 1H), 4.32-4.36 (m, 3H), 6.99-7.40 (m, 4H), 7.29 (dt, J = 7.6, 1.2 Hz, 2H), 7.37 (t, J = 7.6 Hz, 2H), 7.64 (d) , J = 7.6 Hz, 2H), 7.78 (d, J = 7.2 Hz, 2H); MS (ESI) m / z 1050 (M+H) ⁺

Step 2: Preparation of compound 16e

Compound 15e (63 mg, 0.47 mmol) obtained in step 1 was dissolved in MeOH (3 mL), Palladium on carbon (10 wt. % loading, 3 mg, 0.0023 mmol) was added, and hydrogen gas was filled in a reaction vessel at room temperature. stirred for 1 hour. The reaction solution was filtered using Celite, the solvent was removed under reduced pressure, the concentrate was separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 16e (60 mg, 97%).

MS (ESI) m / z 1327 (M+H) ⁺

Step 3: Preparation of compound 17e

Compound 16e (60 mg, 0.045 mmol) obtained in step 2 was dissolved in TFA:TIS:H ₂ O (95:2.5:2.5) solution (2 mL) and stirred at room temperature for 2 hours. After precipitation by adding diethyl ether (20 mL), the mixture was separated using a centrifuge. The crude compound was separated by HPLC, and the purified solution was lyophilized to obtain a solid compound 17e (38 mg, 93%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.30-1.48 (m, 2H), 1.49-1.57 (m, 2H), 1.58-1.82 (m, 5H), 1.83-1.93 (m, 3H), 1.94 -2.06 (m, 1H), 2.08-2.23 (m, 3H), 2.28 (s, 3H), 2.48 (t, J = 6.8 Hz, 2H), 2.57 (t, J = 8.0 Hz, 2H), 2.72- 3.29 (m, 18H), 3.40 (t, J = 4.8 Hz, 2H), 3.53-3.58 (m, 3H), 3.67 (t, J = 4.8 Hz, 2H), 3.71-4.12 (m, 4H), 4.19 -4.22 (m, 1H), 4.28-4.34 (m, 1H), 7.04-7.09 (m, 4H); MS (ESI) m / z 907 (M+H) ⁺

Step 4: Preparation of compound 2e

Compound 17e (10 mg, 0.011 mmol) obtained in step 3 was dissolved in DMF (0.5 mL), DIEA (6 μL, 0.033 mmol) was added, and compound 13 obtained in Preparation Example 1 (5.3 mg, 0.017 mmol) was dissolved in DMF (0.5 mL), added to the reaction solution, and stirred at room temperature for 2 hours. The reaction solution was filtered, diluted with water, separated by HPLC, and the purified solution was freeze-dried to obtain a solid compound 2e (9 mg, 73%).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.31-1.46 (m, 2H), 1.47-1.56 (m, 2H), 1.57-1.81 (m, 5H), 1.82-1.92 (m, 3H), 1.94 -2.08 (m, 1H), 2.09-2.22 (m, 3H), 2.28 (s, 3H), 2.42-2.49 (m, 2H), 2.51-2.59 (m, 4H), 2.62-2.65 (m, 2H) , 2.78-2.96 (m, 3H), 3.00 (s, 3H), 3.02-3.11 (m, 10H), 3.34-3.45 (m, 3H), 3.46-3.59 (m, 5H), 3.64-3.89 (m, 5H), 3.92-4.10 (m, 2H), 4.20-4.26 (m, 1H), 4.31-4.40 (m, 1H)m 4.93 (s, 2H), 7.06 (dd, J = 10.0, 8.8 Hz, 4H) ; MS (ESI) m / z 1114 (M+H) ⁺

<Example 11> Preparation of Ga-chelated compound

<Example 11-1> Preparation of compound 1a

Compound 2a (3 mg, 0.004 mmol) obtained in Example 6 was dissolved in water (0.3 mL), and GaCl ₃ (20 mg, 0.11 mmol) was dissolved in water (0.3 mL) and added thereto, followed by 70 ^o for 10 minutes C was stirred. After confirming the reaction by HPLC, the reaction solution was filtered and separated using HPLC. The purified solution was lyophilized to obtain solid compound 1a (2 mg, 61%).

¹ H NMR (400 MHz, D ₂ O) δ 2.06-2.16 (m, 1H), 2.29-2.42 (m, 1H), 2.54-2.63 (m, 3H), 2.65-2.76 (m, 3H), 2.88- 2.99 (m, 1H), 3.00-3.18 (m, 6H), 3.22-3.31 (m, 2H), 3.32-3.41 (m, 7H), 3.42-3.54 (m, 5H), 3.56-3.60 (m, 4H) ), 3.61-3.67 (m, 1H), 3.74-3.84 (m, 4H), 3.91 (s, 2H), 3.95 (s, 2H), 4.98 (s, 2H); MS (ESI) m / z 850 (M+H) ⁺

<Example 11-2> Preparation of compound 1b

Solid Compound 1b (1 mg, 52%) in the same manner as in Example 11-1, except that Compound 2b (1.8 mg, 0.002 mmol) and GaCl ₃ (11 mg, 0.06 mmol) obtained in Example 7 were used. got

¹ H NMR (400 MHz, D ₂ O) δ 1.55-1.69 (m, 2H), 1.70-1.81 (m, 1H), 1.82-1.91 (m, 1H), 1.98-2.01 (m, 1H), 2.03- 2.16 (m, 2H), 2.32-2.44 (m, 2H), 2.57 (t, J = 6.4 Hz, 2H), 2.60-2.66 (m, 1H), 2.69 (t, J = 7.2 Hz, 2H), 2.90 -3.01 (m, 1H), 3.04 (s, 3H), 3.05-3.15 (m, 2H), 3.20 (t, J = 6.0 Hz, 2H), 3.24-3.70 (m, 18H), 3.74-3.89 (m) , 3H), 3.90-4.02 (m, 2H), 4.30-4.36 (m, 1H), 4.99 (s, 1H); MS (ESI) m / z 949 (M+H) ⁺

<Example 11-3> Preparation of compound 1c

Solid compound 1c (3.5 mg, 66%) was prepared in the same manner as in Example 11-1, except that Compound 2c (5 mg, 0.005 mmol) and GaCl₃ (3 mg, 0.015 mmol) obtained in Example 8 were used. got it

MS (ESI) m/z 1081 [M+H] ⁺

<Example 11-4> Preparation of compound 1d

Solid Compound 1d (1.4 mg, 44%) in the same manner as in Example 11-1, except that Compound 2d (3 mg, 0.003 mmol) and GaCl ₃ (15 mg, 0.08 mmol) obtained in Example 9 were used. got

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 1.34-1.46 (m, 2H), 1.48-1.53 (m, 2H), 1.61-1.73 (m, 2H), 1.74-1.81 (m, 1H), 1.83 -1.97 (m, 4H),, 2.01-2.16 (m, 3H), 2.19 (t, J = 7.6 Hz, 2H), 2.28 (s, 3H), 2.33-2.42 (m, 4H), 2.52-2.67 ( m, 9H), 2.94-3.06 (m, 4H), 3.16-3.19 (m, 3H), 3.24-3.26 (m, 4H), 3.35-3.39 (m, 4H), 3.44-3.58 (m, 6H), 3.67-3.71 (m, 4H), 4.23 (dd, J = 8.4, 4.4 Hz, 1H), 4.35 (dd, J = 9.2, 5.2 Hz, 1H), 4.94 (s, 2H), 7.06 (s, 4H) ; MS (ESI) m / z 1154 (M+H) ⁺

<Example 11-5> Preparation of compound 1e

Solid Compound 1e (1 mg, 39%) in the same manner as in Example 11-1, except that Compound 2e (2.4 mg, 0.002 mmol) and GaCl ₃ (12 mg, 0.07 mmol) obtained in Example 10 were used. made with

¹ H NMR (400 MHz, D ₂ O) δ 1.28-1.40 (m, 2H), 1.44-1.52 (m, 2H), 1.53-1.90 (m, 8H), 1.94-2.06 (m, 2H), 2.21 (t, J = 6.8 Hz, 2H), 2.26 (s, 3H), 2.44-2.60 (m, 5H), 2.65 (t, J = 6.8 Hz, 2H), 2.84-2.97 (m, 2H), 3.01 ( s, 3H), 3.02-3.19 (m, 5H), 3.20-3.64 (m, 18H), 3.67-3.85 (m, 5H), 4.19-4.23 (m, 1H), 4.25-4.29 (m, 1H), 4.95 (s, 2H), 7.13 (dd, J = 14.4, 8.8 Hz, 4H); MS (ESI) m / z 1180 (M+H) ⁺

<Example 12> Preparation of compound [ ⁶⁸ Ga] 1

<Example 12-1> Preparation of compound [ ⁶⁸ Ga] 1a

After flowing 0.1 N hydrochloric acid (5 mL) from the ⁶⁸ Ge/ ⁶⁸ Ga generator, 1 mL each was given to each test tube, and ⁶⁸ Ga solutions (15.3 mCi, 2 mL) were transferred to the reaction vessel in two test tubes with the highest radioactivity. Compound 2a (100 μg) obtained in Example 6 was dissolved in a 1 M sodium acetate solution (pH 4.88, 0.4 mL), and then put into a semi-container. After reacting at 80 °C for 10 minutes, cooled to room temperature, filtered, and separated by HPLC. The separated solution was diluted in water (10 mL), passed through a SepPak (C18) cartridge flowed with distilled water (10 mL), and then dried by blowing nitrogen gas. After eluting with ethanol (1 mL) and drying by blowing nitrogen gas at room temperature, compound [ ⁶⁸ Ga] 1a (4.3 mCi, 68%, decay corrected) was obtained.

HPLC conditions

Column: YMC-Triart Prep C18-S (S-10 μm, 12 nm, 250 mm x 10 mm),

Mobile phase: 8% acetonitrile/water (0.1% TFA),

flow rate: 5 mL/min;

UV detector: 220 mm, RI detector: Diode,

Holding time: 8 min.

<Example 12-2> Preparation of compound [ ⁶⁸ Ga] 1b

^{Compound [ _} _ ⁶⁸ Ga] 1b (3.2 mCi, 49%, decay corrected) was obtained.

HPLC conditions

Column: YMC-Triart Prep C18-S (S-10 μm, 12 nm, 250 mm x 10 mm),

Mobile phase: 9% acetonitrile/water (0.1% TFA),

flow rate: 5 mL/min; UV,

UV detector: 230 mm, RI detector: Diode,

Holding time: 10 min.

<Example 12-3> Preparation of compound [ ⁶⁸ Ga] 1c

^{Compound [ _} _ ⁶⁸ Ga] 1c (3.0 mCi, 49%, decay corrected) was obtained.

HPLC conditions

Column: YMC-Triart Prep C18-S (S-10 μm, 12 nm, 250 mm x 10 mm),

Mobile phase: 27% acetonitrile/water (0.1% TFA),

flow rate: 5 mL/min;

UV detector: 220 mm, RI detector: Diode,

hold time, 12 min.

<Example 12-4> Preparation of compound [ ⁶⁸ Ga] 1d

^{Compound [ _} _ ⁶⁸ Ga] 1d (1.4 mCi, 28%, decay corrected) was obtained.

HPLC conditions

Column: YMC-Triart Prep C18-S (S-10 μm, 12 nm, 250 mm x 10 mm),

Mobile phase: 28% acetonitrile/water (0.1% TFA),

flow rate: 5 mL/min;

UV detector: 230 mm, RI detector: Diode,

Holding time: 11 min.

<Example 12-5> Preparation of compound [ ⁶⁸ Ga] 1e

^{Compound [ _} _ ⁶⁸ Ga] 1e (0.9 mCi, 60%, decay corrected) was obtained.

HPLC conditions

Column: YMC-Triart Prep C18-S (S-10 μm, 12 nm, 250 mm x 10 mm),

Mobile phase: 26% acetonitrile/water (0.1% TFA),

flow rate: 4 mL/min;

UV detector: 230 mm, RI detector: Diode,

Holding time: 16 min.

<Experimental Example 1> logP measurement

The logP value is a measure of fat solubility, and the logP value was measured to evaluate the fat solubility of the radioisotope-labeled compound according to the present invention.

Specifically, after removing the solvent by taking a portion of each solution containing [ ⁶⁸ Ga] 1a -[ ⁶⁸ Ga] 1d obtained in Example 12, PBS buffer (pH 7.4) and n-octanol were added 0.5 mL each After adding, the mixture was mixed with vortex for 1 minute, and then left for 10 minutes to separate the layers. The radiation dose was measured by taking 700 μL from each layer, and the above experiment was repeated 3 times to obtain the logP value as shown in [Table 1] below.

[ ⁶⁸ Ga] logP of 1 [ ⁶⁸ Ga] 1a [ ⁶⁸ Ga] 1b [ ⁶⁸ Ga] 1c [ ⁶⁸ Ga] 1d logP -3.22±0.14 -2.68±0.11 -2.29±0.14 -1.97±0.048

As shown in Table 1, the compound of the present invention has a negative logP value, indicating that the lipophilicity is weak and the hydrophilicity is strong. From this, the compound represented by Formula 1 of the present invention overcomes the problem of high intake in normal organs and low intake in diseased tissues due to the lipophilicity of the radioactive isotope-labeled ligand in the case of the conventional pre-targeting method, By lowering the intake of normal organs and significantly improving the intake of diseased tissues, it has a very high diagnostic and therapeutic effect.

<Experimental Example 2> MicroPET image experiment

In order to evaluate the labeling efficiency and removal rate of the radioisotope-labeled compound according to the present invention, lMicroPET imaging experiments were performed.

Specifically, about 5.0 - 7.4 MBq (200 μL) of [ ⁶⁸ Ga] 1a - [ ⁶⁸ Ga] 1e obtained in Example 12 to male BALB / c mice (8 weeks old, weight 20 g-25 g) through the tail vein, respectively After injection, PET/CT images were obtained for 60 minutes using Inveon™ MicroPET/CT ((Siemens Medical Solutions, Inc, Knoxville, TN). Image results were analyzed using Inveon™ Research Workplace (IRW) program. The results are shown in Figs. 1 to 5. In addition, a graph obtained by quantitative analysis of the values is shown in Fig. 6 .

1 and 2 are MicroPET/CT images of [ ⁶⁸ Ga] 1a and [ ⁶⁸ Ga] 1b , which are compounds without albumin conjugate, and it can be seen that they are rapidly removed through the kidney-bladder.

On the other hand, FIGS. 3 to 5 are MicroPET/CT images of [ ⁶⁸ Ga] 1c , [ ⁶⁸ Ga] 1d and [ ⁶⁸ Ga] 1e , which are compounds containing albumin conjugates, which remain considerably in the blood and are gradually removed through the kidney-bladder it can be seen that

From this, it can be seen that the compound represented by Formula 1 of the present invention includes an albumin conjugate, thereby binding to albumin in the blood, and maintaining the concentration in the blood for a relatively long time.

In addition, it can be seen that the time until complete removal is different depending on the type of the albumin conjugate, so that the retention time can be adjusted according to the size of the binding force with the albumin.

Therefore, if the albumin conjugate having a low binding force with albumin is removed relatively quickly and can be used for diagnosis, if it has an albumin conjugate having a high binding force with albumin, it can be maintained in the blood for a long time and used for treatment.

On the other hand, from the results of FIGS. 1 to 6 , although there is a difference in the residence time in the blood, non-specific binding to normal organs is not observed, so it can be seen that it can specifically act only on diseased organs.

On the other hand, the compound represented by Formula 1 according to the present invention can be formulated in various forms depending on the purpose. The following exemplifies some formulation methods containing the compound represented by Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Formulation Example 1> Preparation of pharmaceutical formulations

1-1. Preparation of powders

500 mg of compound of formula 1

Lactose 100 mg

talc 10 mg

The above ingredients are mixed and filled in an airtight bag to prepare a powder.

1-2. manufacture of tablets

500 mg of compound of formula 1

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets are prepared by tableting according to a conventional manufacturing method of tablets.

1-3. Capsule preparation

500 mg of compound of formula 1

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled in a gelatin capsule to prepare a capsule.

1-4. manufacture of injections

500 mg of compound of formula 1

Appropriate amount of sterile distilled water for injection

Appropriate amount of pH adjuster

According to a conventional method for preparing injections, the content of the above ingredients per 1 ampoule (2 ml) is prepared.

1-5. Preparation of liquids

100 mg of compound of formula 1

10 g isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to a conventional liquid preparation method, each component is added to purified water to dissolve, an appropriate amount of lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. Sterilize to prepare a liquid.

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

Claims (22)

A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Formula 1]

(In Formula 1,
A is -CH- or N;
R ^a is a metal radioisotope-labeled chelator,
The chelator is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or,

ego;
L ¹ , L ² and L ³ are independently spacers,
The spacer is a member; or amino acids, straight or branched C1-20 alkylene, straight or branched C2-20 alkenylene, straight or branched C2-20 alkynylene, -O-, -S-. -S(=O)- -SO2-, -NH-, -N=, -C(=S)-, -C(=O)- and one or more linkers selected from the group consisting of C6-10 arylene a linker consisting of a combination of these;
Tz is tetrazinyl; and
Albumin binder (albumin binder) is an albumin binding residue,
The albumin binding moiety is C6-10 aryl or 5 to 10 membered heteroaryl containing at least one N, and the aryl and heteroaryl are halogen, straight or branched C1-5 alkyl, phenyl and N, O and may be substituted with one or more substituents selected from the group consisting of 5 or 6 membered heteroaryl containing one or more heteroatoms selected from the group consisting of S). The method of claim 1,
The metal radioactive isotope is a positron emitting isotope, a single photon emitting isotope, an Auger electron emitting isotope, a beta particle emitting isotope, an alpha particle emitting isotope, or an isotope emitting two or more radiations. A compound or a pharmaceutically acceptable salt thereof, characterized in that.
The method of claim 1,
The metal radioisotope is Sc-43, Sc-44, Sc-47, Cu-62, Cu-64, Cu-67, Ga-67, Ga-68, Cr-51, Y-88, Y-90, Ru-97, Ru-103, Tc-99m, Rh-105, Pd-109, Sn-117m, In-111, La-140, Ce-141, Pm-149, Tb-152, Tb-152, Tb- 149, Tb-155, Tb-161, Sm-153, Ho-166, Dy-165, Dy-166, Tm-167, Yb-168, Yb-175, Lu-177, Re-186, Re-188, Au-198, Au-199, Pb-203, At-211, Bi-211, Bi-212, Bi-213, Bi-214, Ra-223, Ac-225, Th-227, Th-231, and Th -234 compound or a pharmaceutically acceptable salt thereof, characterized in that any one selected from the group consisting of.
delete delete delete delete According to claim 1,
The tetrazinyl is

and R ¹ is hydrogen, C _1-20 aliphatic hydrocarbon, C _6-10 aryl, or 5 to 10 membered heteroaryl including one or more heteroatoms selected from the group consisting of N, O and S,
Here, the carbon of the aliphatic hydrocarbon may be replaced with one or more heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur,
The aliphatic hydrocarbon is a compound or a pharmaceutically acceptable salt thereof, characterized in that it may be in a saturated or unsaturated state.
delete delete According to claim 1,
the albumin binding moiety is phenyl, naphthalenyl or pyridinyl, wherein the phenyl, naphthalenyl and pyridinyl are selected from the group consisting of halogen, straight or branched C _1-5 alkyl, phenyl and N, O and S A compound or a pharmaceutically acceptable salt thereof, characterized in that it may be substituted with one or more substituents selected from the group consisting of 5 or 6 membered heteroaryl containing one or more heteroatoms.
According to claim 1,
The compound represented by Formula 1 is any one of the compounds consisting of the following compound groups, and M in the following compounds is a metal radioisotope or a pharmaceutically acceptable salt thereof:
(3)

;
(4)

; and
(5)

.
A compound represented by the following formula (2) or a pharmaceutically acceptable salt thereof:
[Formula 2]

(In Formula 2,
A, L ¹ , L ² , L ³ , Tz and the Albumin binder are as defined in Formula 1 of claim 1;
R ^b is a chelator defined in Formula 1 of claim 1).
14. The method of claim 13,
The compound represented by Formula 2 is a compound or a pharmaceutically acceptable salt thereof, characterized in that it is any one of the compounds consisting of the following compound groups:
(3)

;
(4)

; and
(5)

.
A composition for diagnosis of cancer or inflammatory disease, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
16. The method of claim 15,
The composition for diagnosis, characterized in that the albumin-binding moiety of the compound binds to albumin in the blood.
16. The method of claim 15,
Tz of the compound is a diagnostic composition, characterized in that TCO (trans-cyclooctene) and IeDDA (Inverse electron demand Diels-Alder reactions) reaction.
16. The method of claim 15,
The diagnostic composition is a diagnostic composition, characterized in that applicable to a pretargeting (pretargeting) method.
A pharmaceutical composition for the treatment of cancer or inflammatory diseases, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
20. The method of claim 19,
An albumin-binding moiety of the compound binds to albumin in blood.
20. The method of claim 19,
Tz of the compound is TCO (trans-cyclooctene) and IeDDA (Inverse electron demand Diels-Alder reactions) A pharmaceutical composition, characterized in that the reaction.
20. The method of claim 19,
The pharmaceutical composition is a pharmaceutical composition, characterized in that applicable to the pretargeting (pretargeting) method.

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