Novel compound and use thereof

Abstract

To provide a novel compound and use thereof.SOLUTION: A compound is used that comprises: a nuclear translocation unit having the property of migrating into a nucleus of a cell; a functional unit having a specific structure; and a linker unit that links the nuclear translocation unit and the functional unit .SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a novel compound and its use. [Background technology]

[0002] Until now, nuclear medicine therapy has mainly used nuclides that emit beta rays. In recent years, advances in drug delivery technology have made it possible to deliver drugs more precisely to the treatment site, and as a result, clinical applications of alpha rays, which have a shorter radiation range than beta rays and a greater linear energy transfer rate, have been promoted.

[0003] Auger electrons have an even shorter range than alpha rays, so in order to damage therapeutic target cells (e.g., cancer cells), it is thought that a drug that emits Auger electrons must be delivered into the nucleus of the therapeutic target cells. On the other hand, it is expected that damage to cells other than the therapeutic target cells that have taken up the drug can be suppressed. Therefore, attempts are being made to develop various compounds that will enable nuclear medicine therapy using Auger electrons.

[0004] For example, Auger electrons are emitted 125 A compound has been developed in which I-labeled BODIPY (boron dipyrromethene) is bound to Hoechst (see, for example, Non-Patent Documents 1 and 2). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Ryotaro Onoue et. al., "Hoechst-tagged radioiodinated BODIPY derivative for Auger-electron cancer therapy" Chem. Commun., 2023, 59(7), 928-931 [Non-patent document 2] Ryotaro Onoue et. al., "Synthesis and biological evaluation of bimodal BODIPY-conjugated Hoechst applicable for Auger-electron and photodynamic cancer therapy" Bioorg. Med. Chem. Lett., 2023, 96, 129534 Summary of the Invention [Problem to be solved by the invention]

[0006] However, the above-mentioned conventional compounds are not sufficient for clinical application, and the development of novel compounds is desired.

[0007] An object of one aspect of the present invention is to provide a novel compound and use thereof. [Means for solving the problem]

[0008] The present inventors 77 The present inventors have found that a compound having a Br-labeled functional unit having a specific structure, a nuclear transport unit, and a linker unit connecting the nuclear transport unit and the functional unit has excellent in vivo stability, intracellular and nuclear uptake, and cytotoxicity, and have completed the present invention.

[0009] [1] A compound having a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit:

[0010] [ka]

[0011] (In the above formula (1), R1 and R 2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.

[0012] [2] A pharmaceutical composition comprising, as an active ingredient, a compound having a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit:

[0013] [ka]

[0014] (In the above formula (1), R 1 and R 2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.

[0015] [3] The pharmaceutical composition according to [2], wherein the pharmaceutical composition is for use in nuclear medicine therapy and / or photodynamic therapy.

[0016] [4] The pharmaceutical composition according to [2] or [3], wherein the pharmaceutical composition is for treating cancer. [Effects of the Invention]

[0017] According to one aspect of the present invention, a novel compound and its use can be provided. [Brief explanation of the drawings]

[0018] [Figure 1] 1 is a graph showing the test results of stability evaluation of compounds synthesized in the examples of the present invention. [Figure 2]1 is a graph showing the test results of evaluating the uptake of synthesized compounds into cells and nuclei in an example of the present invention. [Figure 3] 1 is a graph showing the test results of evaluating the cytotoxicity of compounds synthesized in the examples of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0019] One embodiment of the present invention will be described below, but the present invention is not limited thereto. The present invention is not limited to the respective configurations described below, and various modifications are possible within the scope of the claims. Embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention. Furthermore, all academic literature and patent documents described in this specification are incorporated herein by reference. Furthermore, unless otherwise specified in this specification, "A to B" representing a numerical range means "greater than or equal to A and less than or equal to B."

[0020] [1. Compound] A compound according to one embodiment of the present invention comprises a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit:

[0021] [ka]

[0022] (In the above formula (1), R 1 and R 2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.

[0023] An embodiment of the present invention has the following advantages (a) to (f): (a) The range of X-rays in tissue is several centimeters or more, the range of β-rays in tissue is 2 to 10 mm, and the range of α-rays in tissue is 28 to 100 μm. On the other hand, the range of Auger electrons in tissue is less than 0.5 μm. One embodiment of the present invention uses a compound that emits Auger electrons, which have an extremely short range, and therefore can specifically damage target cells that have taken up the compound (drug) and prevent damage to non-target cells that have not taken up the compound (drug).

[0024] (b) X-rays have a low linear energy transfer (LET), with β-rays having a linear energy transfer of 0.1 to 10 keV / μm and α-rays having a linear energy transfer of 50 to 230 keV / μm. On the other hand, Auger electrons have a linear energy transfer of 4 to 26 keV / μm. One embodiment of the present invention uses a compound that emits Auger electrons with a sufficiently large linear energy transfer, and therefore can sufficiently damage target cells that have taken up the compound (drug).

[0025] (c) 77 Br can be produced by a small medical cyclotron. 77 Since a compound is used that has a functional unit labeled with Br and having a structure derived from BODIPY, a nuclear transport unit, and a linker unit that connects the nuclear transport unit and the functional unit, the compound can be easily produced in an environment where a small medical cyclotron is installed.

[0026] (d) One embodiment of the present invention is 77 This method uses a compound that includes a Br-labeled functional unit having a BODIPY-derived structure, a nuclear localization unit, and a linker unit that connects the nuclear localization unit to the functional unit. The compound translocates into the nucleus of a target cell by the function of the nuclear localization unit and irradiates the genome in the nucleus with Auger electrons from an extremely short distance. Therefore, one embodiment of the present invention can efficiently damage target cells that have taken up the compound (drug).

[0027] (e) One embodiment of the present invention is 77 A functional unit having a structure derived from BODIPY and labeled with Br is used. Because the functional unit has a structure derived from BODIPY, reactive oxygen species can be generated by irradiating the functional unit with light. Therefore, one embodiment of the present invention can be used in nuclear medicine therapy and / or photodynamic therapy. For example, the combined use of nuclear medicine therapy and photodynamic therapy can more effectively damage target cells.

[0028] (f) 125 I has a long half-life (about 60 days) and remains in the body for a long period of time. 125 I is structurally unstable and is easily released from the labeled compound. 125 I can accumulate in the thyroid gland and cause nonspecific exposure. 77 Br has a moderate half-life (approximately 57 hours) and does not remain in the body for a long period of time. 77 Br is 125 It is structurally more stable than I and is difficult to be released from the labeled form. 77 Even if Br is released, there is no concern that non-specific exposure will occur. Therefore, one embodiment of the present invention is 125 Compared to the technology using I, it is possible to provide a technology with extremely high safety.

[0029] The functional unit, nuclear transport unit, and linker unit are each explained below.

[0030] [1-1. Functional Unit] The functional unit is represented by the following formula (1) or a derivative thereof:

[0031] [ka]

[0032] (In the above formula (1), R 1 and R2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.) In the above formula (1), the functional unit and the linker unit can be bonded via the position indicated by the wavy line.

[0033] The functional unit emits Auger electrons. 77 The functional unit has a structure derived from Br and BODIPY, which generates reactive oxygen species when exposed to light. Therefore, the functional unit can enable nuclear medicine therapy and / or photodynamic therapy.

[0034] In the above formula (1), R 1 and R 2 At least one of 77 Br. Specifically, (i) R 1 but 77 Br and R 2 may be any functional group, and (ii) R 1 is any functional group, and R 2 but 77 Br; and (iii) R 1 and R 2 Both 77 It may also be Br.

[0035] In the above formula (1), R 1 and R 2 may be any functional group. Examples of the functional group include, but are not limited to, H, a sulfo group, a hydroxyl group, a carbonyl group, an acetyl group, an alkyl group, an alkoxy group, an alkylthio group, an aryl group, an aryloxy group, an arylthio group, an arylalkyl group, an arylalkoxy group, an arylalkylthio group, an arylalkenyl group, an arylalkynyl group, an allyl group, an amino group, a substituted amino group, a silyl group, a substituted silyl group, a silyloxy group, a substituted silyloxy group, an arylsulfonyloxy group, an alkylsulfonyloxy group, and a nitro group.

[0036] The functional unit may be a derivative of the functional unit represented by formula (1). The derivative is a derivative resulting from the substitution of a part of the functional unit represented by formula (1) with another functional group or another atom, and has at least one 77 Any derivative containing Br may be used, and the specific structure is not limited.

[0037] Examples of the other functional groups include a sulfo group, a hydroxyl group, a carbonyl group, an acetyl group, an alkyl group, an alkoxy group, an alkylthio group, an aryl group, an aryloxy group, an arylthio group, an arylalkyl group, an arylalkoxy group, an arylalkylthio group, an arylalkenyl group, an arylalkynyl group, an allyl group, an amino group, a substituted amino group, a silyl group, a substituted silyl group, a silyloxy group, a substituted silyloxy group, an arylsulfonyloxy group, an alkylsulfonyloxy group, and a nitro group. Examples of the other atoms include a carbon atom, a hydrogen atom, an oxygen atom, a nitrogen atom, a sulfur atom, a phosphorus atom, and a halogen atom.

[0038] [1-2. Nuclear Import Unit] The nuclear transport unit has the property of transporting into the nucleus of a cell.

[0039] The nuclear transport unit may be any unit that has the property of transporting into the nucleus of a cell, and the specific configuration is not limited.

[0040] Examples of the above-mentioned transfer units include Hochest (e.g., Hochest 33258, Hochest 33342, Hochest 34580), DAPI (4',6-diamidino-2-phenylindole), ethidium bromide, propidium iodide, acridine orange, and derivatives of these compounds. Taking into consideration the ability to accumulate in the nucleus and / or cell membrane permeability, it is preferable to select a compound (transfer unit) with an appropriate molecular weight and / or lipid solubility.

[0041] The derivatives of the nuclear transport unit may be derivatives resulting from the substitution of a portion of the compound with other functional groups or other atoms. Note that the specific structures of the other functional groups and atoms in the nuclear transport unit are the same as those of the other functional groups and atoms in the functional units described in [1-1. Functional Unit], and therefore will not be described here.

[0042] More specifically, the nuclear transport unit may be one represented by the following formula (2) or a derivative thereof. In the following formula (2), the nuclear transport unit and the linker unit may be bonded via the position indicated by the wavy line:

[0043] [ka]

[0044] [1-3. Linker Unit] The linker unit connects the nuclear transport unit and the functional unit.

[0045] The linker unit may be any unit capable of linking the nuclear transport unit and the functional unit, and the specific configuration of the linker unit is not limited. The configuration of the linker unit can be appropriately selected from the viewpoint of the synthesis procedure of the compound according to one embodiment of the present invention and / or the viewpoint of facilitating the synthesis of the compound according to one embodiment of the present invention.

[0046] The linker unit may be, for example, (i) one having 1 to 30, 1 to 20, 1 to 15, or 10 to 15 carbon atoms; (ii) one having one or more ring structures (e.g., a 3- to 6-membered ring, a 4- to 6-membered ring, a 5- to 6-membered ring, or a 6-membered ring); (iii) one having a functional group (e.g., -NH- and / or -CO-); or (iv) any combination of the above (i) to (iii).

[0047] More specifically, the linker unit may be one represented by the following formula (3) or a derivative thereof. In the following formula (3), the nuclear transport unit and the linker unit may be bonded via the position indicated by two wavy lines (for example, the wavy line on the left side of formula (3)), and the functional unit and the linker unit may be bonded via the position indicated by two wavy lines (for example, the wavy line on the right side of formula (3)):

[0048] [ka]

[0049] The derivative of the linker unit may be a derivative obtained by substituting a part of the linker unit represented by formula (3) with another functional group or another atom. Note that the specific configurations of the other functional group and the other atom in the linker unit are the same as the specific configurations of the other functional group and the other atom in the functional unit described in [1-1. Functional Unit], and therefore the description thereof will be omitted here.

[0050] More specifically, the compound according to one embodiment of the present invention may be represented by the following formula (4) or a derivative thereof:

[0051] [ka]

[0052] (In the above formula (4), R 1 and R 2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.) Note that R 1 and R 2 The specific configuration of has been explained in the above section [1-1. Functional Unit], so the explanation will be omitted here.

[0053] The derivative of the compound according to one embodiment of the present invention may be a derivative obtained by substituting a part of the compound represented by formula (4) with another functional group or another atom. Note that the specific configurations of the other functional group and the other atom in the compound according to one embodiment of the present invention are the same as the specific configurations of the other functional group and the other atom in the functional unit described in [1-1. Functional unit], and therefore the description thereof will be omitted here.

[0054] 2. Pharmaceutical Compositions A pharmaceutical composition according to one embodiment of the present invention comprises, as an active ingredient, a compound having a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit:

[0055] [ka]

[0056] (In the above formula (1), R 1 and R 2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.

[0057] The compound according to one embodiment of the present invention has been explained above in [1. Compound], so the explanation thereof will be omitted here.

[0058] The pharmaceutical composition may be for use in nuclear medicine therapy and / or photodynamic therapy.

[0059] The functional unit contained in the compound that is the active ingredient of the pharmaceutical composition emits Auger electrons. 77The pharmaceutical composition has a structure derived from Br and BODIPY, which generates reactive oxygen species when exposed to light. Therefore, the pharmaceutical composition can be used in nuclear medicine therapy and / or photodynamic therapy.

[0060] Recent advances in drug delivery technology have made it possible to deliver drugs to treatment sites. By selecting a known drug delivery technology, the pharmaceutical composition according to one embodiment of the present invention can be delivered to target cells that cause the disease. In other words, a known drug delivery technology can be selected appropriately depending on the type of disease to be treated. Therefore, the diseases that can be treated by the pharmaceutical composition according to one embodiment of the present invention are not limited.

[0061] The pharmaceutical composition can be used, for example, to treat diseases such as cancer.

[0062] The amount of the active ingredient contained in the pharmaceutical composition is not particularly limited, and for example, when the pharmaceutical composition is taken as 100% by mass, it may be 0.00001% by mass to 100% by mass, 0.0001% by mass to 100% by mass, 0.0001% by mass to 100% by mass, 0.001% by mass to 100% by mass, 0.01% by mass to 100% by mass, 0.1% by mass to 100% by mass, 0.1% by mass to 95% by mass, 0.1% by mass to 90% by mass, 0.1% by mass to 80% by mass, 0.1% by mass to 70% by mass, 0.1% by mass to 60% by mass, 0.1% by mass to 50% by mass, 0.1% by mass to 40% by mass, 0.1% by mass to 30% by mass, 0.1% by mass to 20% by mass, or 0.1% by mass to 10% by mass.

[0063] From another perspective, the amount of the active ingredient contained in the pharmaceutical composition is not particularly limited, and may be, for example, 1.0 × 10 -3 ~1.0×10 12 kBq, or 1.0 x 10 -2 ~1.0×10 12 kBq, or 1.0 x 10-1 ~1.0×10 12 kBq, 1.0 to 1.0 × 10 9 kBq, 1.0 to 1.0 × 10 6 kBq, 1.0 to 1.0 × 10 3 It may be kBq.

[0064] The pharmaceutical composition may contain ingredients other than the above-mentioned active ingredients.

[0065] The ingredients other than the active ingredient are not particularly limited and may be, for example, a buffering agent, a pH adjusting agent, an isotonicity agent, a preservative, an antioxidant, a high molecular weight polymer, an excipient, a solvent, an antibacterial agent, or the like.

[0066] Examples of the buffering agent include phosphoric acid or phosphate salts, boric acid or borates, citric acid or citrate salts, acetic acid or acetate salts, carbonic acid or carbonate salts, tartaric acid or tartrate salts, ε-aminocaproic acid, and trometamol. Examples of the phosphate salts include sodium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate. Examples of the borates include borax, sodium borate, and potassium borate. Examples of the citrate salts include sodium citrate, disodium citrate, and trisodium citrate. Examples of the acetate salts include sodium acetate and potassium acetate. Examples of the carbonate salts include sodium carbonate and sodium bicarbonate. Examples of the tartrate salts include sodium tartrate and potassium tartrate.

[0067] Examples of the pH adjuster include hydrochloric acid, phosphoric acid, citric acid, acetic acid, sodium hydroxide, and potassium hydroxide.

[0068] Examples of the isotonic agent include ionic isotonic agents (eg, sodium chloride, potassium chloride, calcium chloride, magnesium chloride) and non-ionic isotonic agents (eg, glycerin, propylene glycol, sorbitol, mannitol).

[0069] Examples of the preservative include benzalkonium chloride, benzalkonium bromide, benzethonium chloride, sorbic acid, potassium sorbate, methyl parahydroxybenzoate, propyl parahydroxybenzoate, and chlorobutanol.

[0070] Examples of the antioxidant include ascorbic acid, tocopherol, dibutylhydroxytoluene, butylhydroxyanisole, sodium erythorbate, propyl gallate, and sodium sulfite.

[0071] Examples of the high molecular weight polymer include methyl cellulose, ethyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose acetate succinate, hydroxypropyl methyl cellulose phthalate, carboxymethyl ethyl cellulose, cellulose acetate phthalate, polyvinylpyrrolidone, polyvinyl alcohol, carboxyvinyl polymer, polyethylene glycol, and atelocollagen.

[0072] Examples of the excipient include lactose, sucrose, D-mannitol, xylitol, sorbitol, erythritol, starch, and crystalline cellulose.

[0073] Examples of the solvent include water, physiological saline, and alcohol.

[0074] Examples of the antibacterial agents include β-lactam, aminoglycoside, tetracycline, lincomycin, chloramphenicol, macrolide, ketolide, polypeptide, and glycopeptide antibiotics; and pyridonecarboxylic acid (quinolone), new quinolone, oxazolidinone, and sulfonamide synthetic antibacterial agents.

[0075] The amount of ingredients other than the active ingredient contained in the pharmaceutical composition is not particularly limited, and may be, for example, 0% to 99.99999% by mass, 0% to 99.9999% by mass, 0% to 99.9999% by mass, 0% to 99.9999% by mass, 0% to 99.999% by mass, 0% to 99.99% by mass, 0% to 99.99% by mass, 5% to 99.9% by mass, 10% to 99.9% by mass, 20% to 99.9% by mass, 30% to 99.9% by mass, 40% to 99.9% by mass, 50% to 99.9% by mass, 60% to 99.9% by mass, 70% to 99.9% by mass, 80% to 99.9% by mass, or 90% to 99.9% by mass.

[0076] The dosage form of the pharmaceutical composition is not limited, and may be, for example, an oral drug, a transdermal drug, an injection, or an infusion.

[0077] The administration interval of the pharmaceutical composition is not limited, and may be, for example, once every hour, once every two hours, once every three hours, once every six hours, once every 12 hours, once every day, once every two days, once every three days, once every four days, once every five days, once every six days, once every week, once every two weeks, once every three weeks, once every month, once every two months, once every three months, once every four months, once every five months, or once every six months.

[0078] The subjects to which the pharmaceutical composition is administered are not limited, and include, for example, humans and non-human organisms (e.g., livestock, pets, and laboratory animals). Non-human organisms include, for example, monkeys, chimpanzees, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, mice, and rats.

[0079] [3. Other] <1> A nuclear medicine therapy and / or photodynamic therapy, comprising a step of administering to a subject (e.g., a human or non-human organism) a pharmaceutical composition containing a compound having a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit:

[0080] [ka]

[0081] (In the above formula (1), R 1 and R 2 teeth, 77 Br or any functional group, and R 1 and R 2 At least one of 77 Br.

[0082] <2> The subject is a cancer patient. <1> Nuclear medicine therapy and / or photodynamic therapy as described in .

[0083] This invention may also contribute to achieving Goal 3 of the United Nations' Sustainable Development Goals (SDGs), including "Ensure good health and promote well-being for all." [Example]

[0084] <1. Compound Synthesis> <1-1. Synthesis of cold form (stable isotope) of compound 7> The cold form of compound 7 was synthesized according to the following scheme.

[0085] [ka]

[0086] With reference to the above-mentioned scheme, the procedure for synthesizing the cold forms of compounds 1 to 7 will be described below.

[0087] (Compound 2) To a THF (27 mL) solution containing compound 1 (0.53 g, 2.80 mmol) and triphenylphosphine (1.43 g, 5.46 mmol), carbon bromide (1.81 g, 5.46 mmol) was added in small portions and allowed to react at room temperature for 1 day. The resulting solution was filtered through Celite, and the resulting residue was separated and purified by silica gel chromatography (chloroform / methanol system) to give compound 2 (0.27 g, 1.07 mmol) in a 38% yield.

[0088] (Compound 4) Potassium carbonate (45 mg, 0.32 mmol) was added to a DMF (1.0 mL) solution containing compound 3 (53 mg, 0.10 mmol) and compound 2 (86 mg, 0.34 mmol), and the mixture was stirred at 60°C overnight. 15 mL of water was added to the reaction solution, and the mixture was extracted with chloroform (15 mL x 3). The resulting extract was dehydrated using sodium sulfate, and then separated and purified by silica gel chromatography (chloroform / methanol system) to obtain compound 4 (6.7 mg, 0.011 mmol) in an 11% yield.

[0089] (Compound 6) Compound 5 (50 mg, 0.11 mmol) was dissolved in 11.7 mL of dehydrated dichloromethane. A solution of N-bromosuccinimide (19 mg, 0.11 mmol) in dehydrated dichloromethane (4.7 mL) was added dropwise to the solution under ice cooling, and the solution was stirred for 30 minutes. The solution was then stirred at room temperature overnight. After removing the reaction solvent under reduced pressure, the resulting reaction product was separated and purified by silica gel chromatography (hexane / ethyl acetate system) to obtain compound 6 (49 mg, 0.90 mmol) in an 82% yield.

[0090] (Compound 7) Trifluoroacetic acid (0.50 mL) was added to an ice-cooled solution of compound 4 (6.7 mg, 0.011 mmol) in dichloromethane (0.75 mL), and the mixture was allowed to react at room temperature for 30 minutes. The solvent was removed from the reaction solution under reduced pressure using an evaporator, and then compound 6 (6.4 mg) in DMF (0.75 mL) and diisopropylethylamine (20 μL) were added to the resulting reaction product and allowed to react overnight at room temperature. 15 mL of water was added to the reaction solution, and the mixture was extracted with chloroform (15 mL × 3). The resulting organic layer was dehydrated using sodium sulfate, and then separated and purified by silica gel chromatography to obtain compound 7 (3.5 mg, 3.8 μmol) in a 34% yield.

[0091] <1-2. Synthesis of the Hot Form of Compound 7 (Radioactive Br-77 Labeled Form)> The Hot form of compound 9 was synthesized according to the following scheme.

[0092] [ka]

[0093] With reference to the above-mentioned scheme, the procedure for synthesizing the Hot form of compound 9 from compound 4 will be described below.

[0094] (Compound 8) Compound 8 was obtained in a 36% yield using Compound 4 and Compound 5 as starting materials in a similar manner to the synthesis method for Compound 7.

[0095] (Radioactive Br) Cyclotron 77 Se(p,n) 77 The Br reaction produces radioactive 77 Br was produced. 77 A tungsten target disk containing Br was heated to 1070°C and subjected to dry and wet distillation. 77 Br was recovered as a 0.5N aqueous ammonia solution.

[0096] (Compound 9) 77 Compound 8 (25 μg), N-chlorosuccinimide (12.5 μg), and 1% acetic acid-methanol solution (50 μL) were added to a vial containing 15.3 MBq of Br evaporated to dryness under reduced pressure. The reaction was carried out at room temperature for 5 minutes with shaking. The reaction solution was then injected into an HPLC column (Nacalai Tesque AR-II 4.6 x 150 mm, Eluent: water-acetonitrile mixed solvent (0.1% TFA), Flow rate: 1.0 mL / min). The peak around 18 minutes after the start of elution was collected to obtain compound 9 (2.6 MBq). The total run time was 23 minutes, with a radiochemical yield of 17% and a radiochemical purity of over 95%. Compound 9 was then solvent-exchanged using a Sep-pak C18 Light column, and used in subsequent experiments. The formation of Compound 9 was confirmed by comparing the retention time of Compound 9 with that of Compound 7 under the same HPLC analysis conditions.

[0097] <2. Physical property evaluation> <2-1. Stability evaluation> Blood collected from mice (BALB / cCrSlc) was stored at 4°C for 16 hours, and then centrifuged (1000 g, 25 minutes) to collect plasma.

[0098] A solution (1.0 MBq, 20 μL) of Compound 9 dispersed in PBS was added to the above plasma (180 μL) and mixed, and the mixture was stored at 37°C.

[0099] The mixture was sampled 5 minutes, 24 hours, and 48 hours after mixing the plasma and the solution. The sampled mixture was injected into HPLC (column: Nacalai Tesque AR-II 4.6 × 150 mm, eluent: water-acetonitrile mixed solvent (0.1% TFA), flow rate: 1.0 mL / min). The radioactivity in the fractions obtained from the peak at 18 minutes after mixing was measured using a gamma counter to evaluate the in vivo stability of Compound 9.

[0100] The test results are shown in Figure 1. As is clear from Figure 1, Compound 9 was extremely stable in vivo. More specifically, Compound 9 was found to be at least 77 It was extremely stable in vivo for a period equivalent to the half-life of Br (approximately 57 hours).

[0101] <2-2. Evaluation of intracellular and intranuclear uptake> The day before the experiment, 1 × 10 HeLa cells were 6 HeLa cells were seeded on a 6-well plate at 1000 cells / well and incubated in 2 mL of DMEM medium (25 mM HEPES, 0.5% EtOH, 0.2% BSA) containing compound 9 (37 kBq).

[0102] Two, four, and six hours after the start of incubation, HeLa cells were harvested using a scraper. The HeLa cells were then collected by centrifugation (2100 g, 3 minutes) and washed with PBS (1 mL x 3). The radioactivity remaining in the cell pellet was measured using a gamma counter to determine the amount of radioactivity incorporated into the cells.

[0103] To the collected cells, 2.0 mL of cell lysis buffer consisting of 10 mM Tris, 1.5 mM MgCl, 140 mM NaCl, and 0.1% IGEPAL-CA630 was added. The mixture was incubated at 0°C for 10 minutes and then centrifuged (1300 g, 2 minutes) to collect the precipitate (nuclear fraction). The radioactivity contained in the collected precipitate was measured using a gamma counter to determine the amount of radioactivity incorporated into the nucleus.

[0104] The test results are shown in Figure 2. In Figure 2, "Intracellular" indicates radioactivity taken up into the cells, and "Nuclear" indicates radioactivity taken up into the nucleus. As is clear from Figure 2, compound 9 was found to have a high ability to migrate into the cells and into the nucleus. More specifically, compound 9 was found to migrate into the nucleus in an extremely short time after contact with the cells.

[0105] Compound 9 and the compounds described in Non-Patent Documents 1 and 2 125 In comparison with the compound labeled with I, compound 9 125 It was superior to compounds labeled with I in that it took a shorter time to translocate into the nucleus after contact with the cell.

[0106] <2-3. Cytotoxicity evaluation> The day before the experiment, 1 × 10 HeLa cells were 4 The HeLa cells were seeded onto a 96-well plate at 100 μL / well. DMEM medium containing 1% EtOH was prepared so that the concentrations of compound 9 were 0 kBq / mL, 1.85 kBq / mL, 3.7 kBq / mL, 18.5 kBq / mL, 37 kBq / mL, 185 kBq / mL, or 370 kBq / mL. 100 μL of this medium was added to each well containing the HeLa cells.

[0107] The HeLa cells were cultured for 2 hours in an incubator maintained at 37°C and 5% CO. Then, 10 μL of MTT solution from an MTT cell count measurement kit manufactured by Nacalai Tesque was added to each well.

[0108] The HeLa cells were cultured for an additional 4 hours. The HeLa cells were then washed with PBS (150 μL × 2), and 100 μL of the solubilization solution included in the kit was added to each well. After confirming that the formazan formed at the bottom of the well had completely dissolved, the absorbance at 570 nm was measured using a plate reader, and the viability of the HeLa cells was calculated based on the absorbance.

[0109] The test results are shown in Figure 3. As is clear from Figure 3, compound 9 reduced the viability of HeLa cells as the radioactivity increased. In other words, compound 9 effectively killed the target cells.

[0110] Compound 9 and the compounds described in Non-Patent Documents 1 and 2 125 When compared with the compound labeled with I, the cytotoxicity was similarly high. This indicates that the use of the present invention can achieve both extremely high safety and extremely high cytotoxicity. [Industrial Applicability]

[0111] The present invention can be used in nuclear medicine therapy and / or photodynamic therapy, and more specifically, in the treatment of various diseases (for example, cancer).

Claims

1. A compound having a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit: 【Chemical 1】 (In the above formula (1), R １ and R ２ teeth, ７７ Br or any functional group, and R １ and R ２ At least one of ７７ Br.).

2. A pharmaceutical composition comprising, as an active ingredient, a compound having a nuclear transport unit capable of transporting into the nucleus of a cell, a functional unit represented by the following formula (1) or a derivative thereof, and a linker unit that connects the nuclear transport unit and the functional unit: 【Chemistry 2】 (In the above formula (1), R １ and R ２ teeth, ７７ Br or any functional group, and R １ and R ２ At least one of ７７ Br.).

3. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition is for use in nuclear medicine therapy and / or photodynamic therapy.

4. The pharmaceutical composition according to claim 2 or 3, which is for treating cancer.

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