Radiolabeled tyrosine derivatives and uses thereof

Abstract

Provided is a radiolabeled tyrosine derivative which has excellent LAT1 selectivity, has higher retention at tumor or cancer sites, has a clearance rate to a degree that does not cause side effects, and can be produced stably with good purity by a safe method suitable for industrial production. The present invention provides a radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof. # imgabs0 # [In the formula, each symbol is as defined in the description].

Description

Technical Field

[0001] The present invention relates to a radioactively labeled tyrosine derivative which can be used as a therapeutic drug for tumors or cancers and its use, as well as a method for producing the tyrosine derivative and a synthetic intermediate thereof. Background Art

[0002] Radiolabeled tyrosine derivatives are known, such as astatine (2 11 At)-α-methyl-L-tyrosine (hereinafter also referred to as 211 At-AAMT) is a drug that is taken up into tumor cells via the amino acid transporter LAT1 (L-type amino acid transporter 1, hereinafter also referred to as LAT1) that is specifically expressed in tumors, and can be used as an anticancer agent (Patent Document 1).

[0003] but, 211 Although At-AAMT has been confirmed to have anti-tumor effects, it is cleared very quickly. Therefore, it is expected to develop compounds with higher retention in tumor or cancer sites. In addition, among the major obstacles in drug development, there is acquired drug resistance, but its original state is considered to be the overexpression of multidrug efflux transporters represented by ABC transporters. Compounds with high retention not only have stronger therapeutic effects at the same dose, but are also expected to be suitable for the treatment of tumors or cancers that have acquired anticancer drug resistance.

[0004] Patent Document 1 discloses that conventionally, α-methyl-L-tyrosine (AMT) is dissolved in sulfuric acid, mercuric sulfate is added to introduce mercury into the benzene ring, and then an astatine exchange reaction is performed to produce AMT. 211 At-AAMT (hereinafter also referred to as mercury method). This method uses mercury, a harmful substance, and therefore cannot be said to be a method suitable for manufacturing pharmaceuticals in terms of safety. In addition, in the above method, since the synthesis yield varies greatly between manufacturing batches, it is not suitable for industrial production. In addition, since by-product iodine substitution and halogen di-substitution are generated, there are problems in purity.

[0005] On the other hand, the inventors of the present invention reported that the boron group (-B(OH)2) after the introduction of the aromatic group has excellent astatine substitution ability (Patent Document 2)

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent document 1: WO2019 / 176505

[0009] Patent Document 2: WO2019 / 027059 Summary of the invention

[0010] Problem that the invention aims to solve

[0011] The present invention aims to provide a radiolabeled tyrosine derivative having excellent LAT1 selectivity, higher retention in tumor or cancer sites, a clearance rate at a level that does not cause side effects, and which can be produced stably and with high purity by a safe method suitable for industrial production.

[0012] Means of solving the problem

[0013] In order to solve the above problems, the present inventors conducted intensive research and found that the novel compound represented by the following formula (I) has excellent LAT1 selectivity, higher retention in tumor or cancer sites, and a clearance rate that does not cause side effects, and thus can exert better anti-tumor effects. In addition, surprisingly, it was also found that the above compound can also exert excellent anti-tumor effects on tumors or cancers that have acquired anticancer drug resistance.

[0014] In addition, the inventors of the present invention have discovered a method for producing a compound represented by the following formula (I) without using harmful substances such as mercury, by introducing a boron group (-B(OH)2) or its ester group as a new intermediate to produce a compound represented by the following formula (II) and a compound represented by the following formula (III).

[0015] As a result, the invention was completed.

[0016] That is, the present invention is as follows.

[0017] [1] a radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof (hereinafter also referred to as radiolabeled compound (I)),

[0018]

[0019] In formula (I), R 1 Represents C 1-4 alkyl;

[0020] R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl;

[0021] R 3 represents a fluorine atom or a chlorine atom;

[0022] m represents 0, 1 or 2;

[0023] n represents 0, 1 or 2;

[0024] X represents the selection 211 At 210At 131 I. 125 I and 77 Radioactive nuclide of Br.

[0025] [2] The compound according to [1] above, or a pharmaceutically acceptable salt thereof, wherein m is 1.

[0026] [3] The compound according to [1] above, or a pharmaceutically acceptable salt thereof, wherein n is 0.

[0027] [4] The compound according to [1] above or a pharmaceutically acceptable salt thereof, wherein R 2 C 1-6 alkyl.

[0028] [5] The compound according to [1] above or a pharmaceutically acceptable salt thereof, wherein R 1 It is methyl.

[0029] [6] The compound according to [1] above or a pharmaceutically acceptable salt thereof, wherein R 2 O is bonded to the 4-position of the benzene ring.

[0030] [7] The compound or pharmaceutically acceptable salt thereof according to [1] above, wherein X is bonded to the 3-position of the benzene ring.

[0031] [8] The compound according to [1] above or a pharmaceutically acceptable salt thereof, wherein X is 211 At.

[0032] [9] A pharmaceutical composition comprising the compound according to any one of [1] to [8] above or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0033]

[10] The pharmaceutical composition according to [9] above, further comprising at least one selected from ascorbic acid, an alkali metal ascorbic acid salt and an alkaline earth metal ascorbic acid salt.

[0034]

[11] A therapeutic agent for a tumor or cancer expressing the amino acid transporter LAT1, comprising the compound according to any one of [1] to [8] above or a pharmaceutically acceptable salt thereof.

[0035]

[12] The therapeutic agent according to the above

[11] , wherein the tumor or cancer expressing the amino acid transporter LAT1 is selected from pancreatic cancer, leukemia, melanoma, colorectal cancer, lung cancer, prostate cancer, gastric cancer, breast cancer, kidney cancer, laryngeal cancer, esophageal cancer, liver cancer, lymphoma, myeloma, head and neck cancer, ovarian cancer, bladder cancer, childhood cancer, childhood leukemia, brain tumor, osteosarcoma, soft tissue sarcoma and bone and soft tissue tumors.

[0036]

[13] The therapeutic agent according to

[11] above, wherein the tumor or cancer is a tumor or cancer that has acquired resistance to an anticancer agent.

[0037]

[14] The therapeutic agent according to

[11] above, wherein the tumor or cancer is a tumor or cancer that has acquired resistance to platinum agents.

[0038]

[15] The therapeutic agent according to

[11] above, wherein the tumor or cancer is a tumor or cancer that has acquired cisplatin resistance.

[0039]

[16] The therapeutic agent according to

[11] above, wherein the tumor or cancer is a tumor or cancer that has acquired resistance to a nucleic acid synthesis inhibitor.

[0040]

[17] The therapeutic agent according to

[11] above, wherein the tumor or cancer is a tumor or cancer that has acquired gemcitabine resistance.

[0041]

[18] a compound represented by formula (II) or a salt thereof (hereinafter also referred to as boronic acid compound (II)),

[0042]

[0043] In formula (II), R 1 Represents C 1-4 alkyl;

[0044] R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl;

[0045] R 3 represents a fluorine atom or a chlorine atom;

[0046] m represents 0, 1 or 2;

[0047] n represents 0, 1 or 2;

[0048] Y represents a boron group (-B(OH)2) or an ester group thereof.

[0049]

[19] The compound or salt thereof according to

[18] above, wherein Y is a boryl group (—B(OH) 2 ) or a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group.

[0050]

[20] A drug for boron neutron capture therapy, comprising the compound described in

[18] or

[19] above or a pharmaceutically acceptable salt thereof.

[0051]

[21] A method for producing a radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof, comprising the following steps:

[0052] Step 1: In the presence of a reagent selected from the group consisting of alkali metal iodides, alkali metal bromides, N-bromosuccinimide, N-chlorosuccinimide, N-iodosuccinimide and hydrogen peroxide, a compound represented by formula (II) or a salt thereof is reacted in water with a 211 At 210 At 131 I. 125 I, and 77 A step of obtaining a radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof by reacting a radionuclide of Br with Br,

[0053]

[0054] In the formula, R 1 Represents C 1-4 alkyl;

[0055] R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl;

[0056] R 3 represents a fluorine atom or a chlorine atom;

[0057] m represents 0, 1 or 2;

[0058] n represents 0, 1 or 2;

[0059] X represents the selection 211 At 210 At 131 I. 125 I and 77 Radionuclides of Br;

[0060] Y represents a boron group (-B(OH)2) or an ester group thereof.

[0061]

[22] a compound represented by formula (III) (hereinafter also referred to as boronic acid compound (III)),

[0062]

[0063] In formula (III),

[0064] R 1 Represents C 1-4 alkyl;

[0065] R 2 Represents C1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl;

[0066] R 3 represents a fluorine atom or a chlorine atom;

[0067] m represents 0, 1 or 2;

[0068] n represents 0, 1 or 2;

[0069] Y represents a boron group (-B(OH)2) or an ester group thereof;

[0070] P 1 represents a carboxyl protecting group;

[0071] P 2 represents an amino protecting group.

[0072]

[23] The compound or salt thereof according to

[22] above, wherein P 1 is benzyl, and P 2 It is tert-butyloxycarbonyl.

[0073]

[24] A compound represented by formula (IV'),

[0074]

[0075] In formula (IV'), R 2a C 1-3 Alkyl, P 1a Represents benzyl or C 1-3 alkyl.

[0076] [24a] A compound represented by formula (IV")

[0077]

[0078] In formula (IV"), P 1a Represents benzyl or C 1-2 alkyl.

[0079] Effects of the Invention

[0080] According to the present invention, a compound having excellent LAT1 selectivity, higher retention in tumor or cancer sites, and clearance without causing side effects can be provided, thereby being able to exert a better anti-tumor effect. In addition, a compound capable of exerting an excellent anti-tumor effect on tumors or cancers that have acquired anticancer drug resistance can also be provided.

[0081] In addition, since the compound is a tyrosine derivative (boronic acid compound (III) and boronic acid compound (II)) into which a boron group (-B(OH)2) or its ester group is introduced, the above-mentioned compound can be produced stably and with good purity by a safe method suitable for industrial production of pharmaceuticals without using harmful substances. BRIEF DESCRIPTION OF THE DRAWINGS

[0082] Figure 1 The HPLC chart of (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-methyl-α-methyltyrosine hydrochloride (6) (Bpin-AMT-OMe) obtained in Example 1 is shown.

[0083] Figure 2 The results obtained in Example 2 are shown in FIG. 211 A graph showing the results of thin layer chromatography (TLC) analysis of a crude At-AAMT-OMe solution.

[0084] Figure 3 The results obtained in Example 2 are shown in FIG. 211 A diagram showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OMe before purification, immediately after production, 3 hours after production, and 24 hours after production.

[0085] Figure 4 The (R)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-methyl-α-methyltyrosine hydrochloride ( 12 HPLC chart of )((R)-Bpin-AMT-OMe).

[0086] Figure 5 is a graph showing the (R)- 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OMe after purification.

[0087] Figure 6 is a graph showing the (R)- 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OMe 24 hours after production.

[0088] Figure 7 The (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-ethyl-α-methyltyrosine hydrochloride ( 18 )HPLC chart of Bpin-AMT-OEt.

[0089] Figure 8 The results obtained in Example 6 are shown in FIG. 211A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OEt after purification.

[0090] Fig. 9 The results obtained in Example 6 are shown in FIG. 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OEt 24 hours after production.

[0091] Fig.10 The (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-propyl-α-methyltyrosine hydrochloride ( 24 )HPLC chart of Bpin-AMT-OPr.

[0092] Fig.11 The results obtained in Example 8 are shown in FIG. 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OPr after purification.

[0093] Fig.12 The results obtained in Example 8 are shown in FIG. 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMT-OPr 24 hours after production.

[0094] Fig.13 The (S)-α-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-methoxyphenylalanine hydrochloride ( 28 HPLC chart of )(Bpin-AMPhe-OMe).

[0095] Fig.14 The results obtained in Example 10 are shown in FIG. 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMPhe-OMe after purification.

[0096] Fig.15 The results obtained in Example 10 are shown in FIG. 211 A graph showing the results of thin layer chromatography (TLC) analysis of At-AAMPhe-OMe 24 hours after production.

[0097] Fig.16 This is a diagram showing the 211 At-AAMT-OMe or 211 Graph showing the uptake of At-AAMT into cells (HEK293).

[0098] Fig.17 This is a graph showing that in Experimental Example 2, PANC1 was subcutaneously transplanted into mice. 211 At-AAMT-OMe or211 Figure 2 shows the in vivo distribution of At-AAMT after 2 hours.

[0099] Fig.18 This is a graph showing that in Experimental Example 3, PANC1 was subcutaneously transplanted into mice. 211 At-AAMT-OMe or 211 Graph of tumor weight 28 days after At-AAMT.

[0100] Fig.19 This is a graph showing that in Experimental Example 3, PANC1 was subcutaneously transplanted into mice and the 211 At-AAMT-OMe or 211 Figure 2 shows the body weight of mice 28 days after At-AAMT administration.

[0101] Fig. 20 It is shown that in Test Example 4 211 At-AAMT-OMe, 211 At-AAMT-OEt, 211 At-AAMT-OPr or 211 Graph of At-AAMT uptake into cells (PANC1).

[0102] Fig.21 This is a graph showing that in Experimental Example 6, PANC1 was subcutaneously transplanted into mice and the 211 At-AAMT-OMe or 211 Figure 3. Tumor size of At-AAMT-OEt at 21 days after administration.

[0103] Fig. 22 This is a graph showing that in Experimental Example 6, PANC1 was subcutaneously transplanted into mice and the 211 At-AAMT-OMe or 211 Figure 2 shows the body weight of mice 21 days after the administration of At-AAMT-OEt.

[0104] Fig.23 This is a graph showing the cell viability of A549 and CDDP-resistant A549 (A549 CDDP-r) in Test Example 7.

[0105] Fig.24 It is shown that in Test Example 8, by applying 211 At-AAMT-OMe or 211 Graph of cell viability of At-AAMT, A549 and CDDP-resistant A549 (A549 CDDP-r).

[0106] Fig.25 This is a diagram showing that the administration of the drug to A549-transplanted mice and CDDP-resistant A549 (A549CDDP-r)-transplanted mice in Experimental Example 9211 At-AAMT-OMe or 211 Graph of tumor weight 30 days after At-AAMT.

[0107] Fig.26 This is a graph showing the expression level of LAT1 on the cell surface of A549 and CDDP-resistant A549 (A549 CDDP-r) in Test Example 10.

[0108] Fig. 27 This is a graph showing the cell survival rates of PANC1 and gemcitabine-resistant PANC1 (PANC1Gem-r) in Test Example 11.

[0109] Fig.28 This is a graph showing that the administration of the drug to PANC1 transplanted mice and gemcitabine-resistant PANC1 (PANC1 Gem-r) transplanted mice in Experimental Example 12 211 Graph of tumor weights of mice 30 days after At-AAMT-OMe.

[0110] Fig.29 This is a graph showing the effect of the PANC1 transplanted mice and gemcitabine-resistant PANC1 (PANC1 Gem-r) transplanted mice immediately after administration of 211 Graph of tumor size in mice 14 days after administration of At-AAMT-OMe.

[0111] Fig.30 This is a graph showing the effect of the PANC1 transplanted mice and gemcitabine-resistant PANC1 (PANC1 Gem-r) transplanted mice immediately after administration of 211 Figure 3. Body weight of mice 14 days after administration of At-AAMT-OMe. DETAILED DESCRIPTION

[0112] Hereinafter, the present invention will be described in detail.

[0113] In the present specification, examples of the "halogen atom" include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

[0114] In this manual, “C 1-4 The "alkyl group" may, for example, be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.

[0115] In this manual, “C 1-6 The “alkyl group” includes, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 1-ethylpropyl, hexyl, isohexyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl and 2-ethylbutyl.

[0116] In this manual, "C 1-6 A "haloalkyl" is a "C" substituted with one or more "halogen atoms". 1-6 The "alkyl group" includes, for example, fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, trichloromethyl, 2-fluoroethyl, 2-bromoethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 2,2-difluoropropyl, 3,3,3-trifluoropropyl, 4,4,4-trifluorobutyl, 5,5,5-trifluoropentyl, 6,6,6-trifluorohexyl, etc.

[0117] In this manual, “C 6-10 The "aryl group" includes, for example, phenyl, 1-naphthyl and 2-naphthyl.

[0118] In this manual, "C 7-14 Arylalkyl" is replaced by "C 6-10 Aryl" substituted "C 1-4 The "alkyl group" includes, for example, benzyl, phenethyl, 2-phenethyl, naphthylmethyl, phenylpropyl and the like.

[0119] The "boryl group (-B(OH)2)" in this specification is also referred to as a dihydroxyboryl group.

[0120] In the present specification, examples of the "boryl ester group" include the following groups.

[0121]

[0122] [Where R 4 Represents C 1-6 alkyl]

[0123] In the present specification, examples of the “amino-protecting group” include tert-butoxycarbonyloxy, benzyloxycarbonyl, and 9-fluorenylmethyloxycarbonyl.

[0124] In the present specification, examples of the "carboxyl protecting group" include benzyl, C 1-2 Alkyl (methyl, ethyl), tert-butyl, etc.

[0125] The radiolabeled compound (I) of the present invention is the compound shown below.

[0126]

[0127] R 1 Represents C 1-4 alkyl.

[0128] As one embodiment, R1 is preferably a methyl group.

[0129] R 2 Represents C 1-6Alkyl (preferably methyl, ethyl, propyl), C 1-6 Haloalkyl (preferably trifluoromethyl, 2-fluoroethyl) or C 7-14 Aralkyl (preferably benzyl).

[0130] As a way, R 2 Preferably, it is methyl, ethyl, propyl, trifluoromethyl, 2-fluoroethyl or C 7-14 The aralkyl group is more preferably a methyl group, an ethyl group, a propyl group or a trifluoromethyl group, further preferably a methyl group, an ethyl group or a propyl group, and particularly preferably a methyl group.

[0131] As another way, R 2 Preferably C 1-6 Alkyl, more preferably C 1-4 The alkyl group is more preferably a methyl group, an ethyl group or a propyl group, and particularly preferably a methyl group.

[0132] R3 represents a fluorine atom or a chlorine atom.

[0133] As a way, R 3 Preferred is a fluorine atom.

[0134] n represents 0, 1, or 2.

[0135] As one aspect, n is preferably 0.

[0136] m represents 0, 1, or 2.

[0137] As one aspect, m is preferably 1.

[0138] X represents the selection 211 At (nuclides emitting alpha rays), 210 At (nuclides emitting alpha rays), 131 I (beta-emitting nuclide), 125 I (X-ray emitting nuclide) and 77 A radionuclide of Br (an Auger electron-emitting nuclide).

[0139] The half-lives of these radionuclides are: 211 At is 7.2 hours, 210 At is 8.3 hours, 131 I is 8.04 days, 125 I is 59.4 days, 77 Br is 57 hours.

[0140] As one approach, X is preferably 211 At.

[0141] R 2 The bonding position of O on the benzene ring is not particularly limited, but is preferably the 3-position or 4-position, and particularly preferably the 4-position.

[0142] The bonding position of X on the benzene ring is not particularly limited. For example, when R 2 When O is bonded to the 4-position of the benzene ring, X is preferably at the 3-position. 2 When O is bonded to the 3-position of the benzene ring, X is preferably at the 4-position.

[0143] R 3 The bonding position on the benzene ring is not particularly limited, but is preferably the 2-position, 5-position or 6-position, and particularly preferably the 5-position.

[0144] The stereo configuration in formula (I) is not particularly limited, and may be any of D-form, L-form, and DL-form (i.e., may be any of R-form, S-form, and R / S-form). That is, the radiolabeled compound (I) may be any of the radiolabeled compound (Ia), compound (Ib), and racemic body represented by the following formula.

[0145]

[0146] [The symbols in the formula are the same as above.]

[0147] As one embodiment, the radiolabeled compound (I) is preferably the radiolabeled compound (Ia) from the viewpoint of the amount taken up into tumor or cancer cells.

[0148] Specific examples of the radiolabeled compound (I) include the following compounds.

[0149]

[0150]

[0151]

[0152]

[0153] [wherein, Me is a methyl group, Et is an ethyl group, Pr is a propyl group, Bn is a benzyl group, and X has the same meaning as above, and is preferably 211 At, m has the same meaning as above, and is preferably 1.]

[0154] The compound represented by formula (I) may be in the form of a pharmaceutically acceptable salt thereof. Pharmaceutically acceptable salts include inorganic salts such as alkali metal salts (e.g., sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g., calcium salts, magnesium salts, barium salts, etc.), ammonium salts, salts with inorganic acids such as hydrogen chloride, hydrogen bromide, nitric acid, sulfuric acid, phosphoric acid, or salts with organic acids such as acetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, etc.

[0155] Next, the method for producing the radiolabeled compound (I) of the present invention is described.

[0156] In the present specification, when the raw material compound is a salt, examples of such salts include metal salts (e.g., alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts, magnesium salts and barium salts), ammonium salts, salts with organic bases (trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine), salts with inorganic acids (e.g., hydrogen chloride, hydrogen bromide, nitric acid, sulfuric acid, phosphoric acid), salts with organic acids (e.g., formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, p-toluenesulfonic acid), and the like.

[0157] The radiolabeled compound (I) can be produced by a method comprising the following step 1.

[0158]

[0159] [In the formula, Y represents a boron group (-B(OH)2) or an ester group thereof, and each symbol has the same meaning as above.]

[0160] In one embodiment, Y is preferably a boron group (—B(OH) 2 ) or a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, and particularly preferably a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group.

[0161] Step 1 is to react a boronic acid compound (II) with a boronic acid compound (II) in water in the presence of a reagent selected from an alkali metal iodide, an alkali metal bromide, N-bromosuccinimide, N-chlorosuccinimide, N-iodosuccinimide and hydrogen peroxide. 211 At 210 At 131 I. 125 I, and 77 A step of obtaining a radiolabeled compound (I) by reacting the radionuclide of Br.

[0162] Boric acid compound (II) is a new type of compound.

[0163] The boronic acid compound (II) may be any of a boronic acid compound (IIa), a compound (IIb), and a racemic form represented by the following formula.

[0164]

[0165] [The symbols in the formula are the same as above.]

[0166] As one embodiment, the boronic acid compound (II) is preferably a boronic acid compound (IIa).

[0167] Specific examples of the boronic acid compound (II) include the following compounds.

[0168]

[0169]

[0170]

[0171]

[0172] [In the formula, Y has the same meaning as above, preferably a boron group (-B(OH)2) or a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, and particularly preferably a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, m has the same meaning as above, preferably 1, and Me, Et, Pr and Bn have the same meaning as above.]

[0173] The boronic acid compound (II) can be produced by the following method.

[0174] Since the reaction in this step is carried out in water, the boric acid compound (II) may be in a free form or in a salt form as long as it is soluble in water. Alternatively, it may be dissolved in a weakly alkaline aqueous solution such as a sodium bicarbonate aqueous solution and used.

[0175] Examples of the alkali metal iodide include potassium iodide and sodium iodide. Among them, potassium iodide is preferably used.

[0176] Examples of the alkali metal bromide include sodium bromide and potassium bromide.

[0177] Preferred combinations of the radionuclide and the above reagent include:

[0178] (1) Radionuclides are 211 At or 210 At, and the above reagent is selected from the combination of potassium iodide, sodium bromide, N-bromosuccinimide, N-chlorosuccinimide, N-iodosuccinimide and hydrogen peroxide;

[0179] (2) Radionuclides are 125 I or 131 I, and the above reagent is selected from the combination of N-bromosuccinimide and N-chlorosuccinimide;

[0180] (3) Radionuclides are 77 Br, and the above reagent is a combination of N-chlorosuccinimide.

[0181] The above reagents may be used alone or in combination of two or more. The above reagents are usually used in the form of aqueous solutions.

[0182] As a preferred embodiment, the radionuclide is 211 At or 131 I, and the reagent is selected from potassium iodide and N-bromosuccinimide.

[0183] As a more preferred embodiment, the radionuclide may be 211 At and the reagent is potassium iodide, and the radionuclide is 131 I and the reagent is N-bromosuccinimide.

[0184] The reagents may be used in an amount capable of oxidizing or reducing the radionuclide and are usually used in a large excess relative to the radionuclide. However, from the viewpoint of reaction efficiency and economic efficiency, the concentration is preferably 0.0001 to 0.2 mol / L, more preferably 0.001 to 0.1 mol / L.

[0185] The radioactive nuclide is usually used in the reaction in an aqueous solution state. If necessary, an alkaline aqueous solution such as sodium hydroxide or a buffer solution may be added to the aqueous solution in order to stabilize the radioactive nuclide.

[0186] When the radionuclide is 211 At 28 MeV helium particles accelerated by a cyclotron are irradiated onto bismuth. 209 Bi(α, 2n) 211 At nuclear reaction to produce 211 After At, the target substance 209 Bi is heated and melted, 211 At evaporates and is collected in a cold trap, which is dissolved in water to prepare 211 At stock solution. As needed, in order to stabilize 211 At, an alkaline aqueous solution such as sodium hydroxide or a buffer solution may also be added.

[0187] When the radionuclide is 210 At, helium particles accelerated to 29 MeV or more by a cyclotron are irradiated onto bismuth. 209 Bi(α, 3n) 210 At nuclear reaction to produce 210 After At, the same operation as above was performed to prepare 210 At aqueous solution.

[0188] When the radionuclide is 125 I, can be used as Na 125 I aqueous solution was obtained.

[0189] When the radionuclide is 131 I, can be used as Na 131 I aqueous solution was obtained.

[0190] When the radionuclide is 77 Br, proton particles accelerated by a cyclotron are irradiated onto tellurium. 77 Se(p,n) 77 Br is produced by nuclear reaction 77 Br, dissolve the target substance 77 Se, preparation 77 Br in sodium hydroxide solution.

[0191] because 211 The half-life of At is as short as 7.2 hours. 210 At has a short half-life of 8.3 hours, so the radionuclide must be used in the reaction immediately after preparation. 125 The half-life of I is 59.4 days. 131 The half-life of I is 8.04 days. 77 Br has a relatively long half-life of 57 hours, but these radionuclides are preferably used in the reaction quickly after preparation.

[0192] The boronic acid compound (II) is usually used in a large excess relative to the radioactive nuclide, but from the viewpoint of reaction efficiency and economic efficiency, it is preferably used at a concentration of 0.00001 mol / L to 0.5 mol / L, more preferably 0.0001 mol / l to 0.2 mol / L relative to 1 Bq to 1000 GBq of the radioactive nuclide.

[0193] The above reaction is carried out by mixing the boronic acid compound (II), the above reagent and the radioactive nuclide, and there is no particular restriction on the order of mixing them. Preferably, the method comprises adding an aqueous solution of radioactive nuclide to an aqueous solution of the boronic acid compound (II) and then adding an aqueous solution of the above reagent, or the method comprises adding an aqueous solution of radioactive nuclide to an aqueous solution of the boronic acid compound (II) and then adding an aqueous solution of the above reagent, and more preferably, the method comprises adding an aqueous solution of radioactive nuclide to an aqueous solution of the boronic acid compound (II) and then adding an aqueous solution of the above reagent.

[0194] The above reaction is carried out in water, that is, in a system without containing an organic solvent.

[0195] The above reaction is carried out at 0 to 95° C., preferably 10 to 80° C. The reaction time is 1 minute to 3 hours, preferably 1 minute to 1 hour.

[0196] The completion of the reaction was confirmed by the disappearance of free radionuclides by thin layer chromatography (TLC) analysis.

[0197] In the production method of the present invention, the radioactive labeled compound (I) can be obtained with a high radiochemical yield of 60% or more, particularly 80% or more, and especially 90% or more.

[0198] After the reaction is completed, since the reaction solution does not contain organic solvents or toxic reagents, the radioactive labeled compound (I) can be directly prepared into an injection or the like without isolating it.

[0199] The reaction between the boronic acid compound (II) and the radioactive nuclide is an electrophilic substitution reaction and / or a nucleophilic substitution reaction. Since the introduction site of the radioactive nuclide in the boronic acid compound (II) is the benzene ring, especially in 211 At or 210 In the case of At, the benzene ring can be introduced well.

[0200] In addition, in order to remove by-products, the radioactive labeled compound (I) can be purified as needed. The purification is preferably carried out using a solid phase extraction column. As the solid phase extraction column, a solid phase extraction column commonly used in the technical field can be used.

[0201] Furthermore, after the above purification, ascorbic acid or an alkali metal salt or alkaline earth metal salt of ascorbic acid may be added to a final concentration of 0.01% to 10%, preferably 0.1% to 5%. This can suppress the decomposition of the radiolabeled compound (I) and preserve it for a long time.

[0202] The boronic acid compound (II) can be produced by a method comprising the following steps 2 to 5.

[0203]

[0204] [wherein, Z is an iodine atom or a bromine atom, P 1 represents a carboxyl protecting group, P 2 represents an amino protecting group, and other symbols have the same meanings as above.]

[0205] As one aspect, Z is preferably an iodine atom.

[0206] As a way, P 1 Preferably benzyl or C 1-2 The alkyl group (methyl group, ethyl group) is more preferably a benzyl group.

[0207] As a way, P 2 Preferred is tert-butoxycarbonyloxy or benzyloxycarbonyloxy, more preferred is tert-butoxycarbonyloxy.

[0208] Step 2 is a step of iodinating or brominating a compound represented by formula (VI) or a salt thereof (hereinafter also referred to as compound (VI)) to obtain a compound represented by formula (V) or a salt thereof (hereinafter also referred to as compound (V)).

[0209] Iodination or bromination can be performed by reacting compound (VI) with an iodinating agent or a brominating agent.

[0210] As compound (VI), α-methyltyrosine, α-methyl-m-tyrosine, 4-hydroxy-α-methylphenylglycine, 3-hydroxy-α-methylphenylglycine, 2-amino-4-(4-hydroxyphenyl)-2-methylbutyric acid, 2-amino-4-(3-hydroxyphenyl)-2-methylbutyric acid, etc. can be listed, and they can be any of L-type, D-type, and DL-type. Among them, α-methyltyrosine and α-methyl-m-tyrosine are preferably used, and α-methyltyrosine is particularly preferably used. Compound (VI) can be manufactured using commercial products or by known methods.

[0211] Examples of the iodination agent include iodine and N-iodosuccinimide.

[0212] Examples of the brominating agent include bromine and N-bromosuccinimide.

[0213] The amount of the iodinating agent or brominating agent used is generally 1 to 5 mol, preferably 1 to 2 mol, based on 1 mol of compound (VI).

[0214] When iodine is used, the reaction is carried out in the presence of potassium iodide and concentrated ammonia (28%). The amount of potassium iodide used is usually 0.5 to 5 moles, preferably 1 to 2 moles, relative to 1 mole of iodine, and the amount of concentrated ammonia used is usually 5 to 200 moles, preferably 20 to 50 moles, relative to 1 mole of iodine.

[0215] The reaction is usually carried out in a solvent. As the solvent used in the present invention, there is no particular limitation as long as it has no adverse effect on the reaction, and examples thereof include water; alcohol solvents such as ethanol, methanol, isopropanol; halogen solvents such as carbon tetrachloride, etc., and two or more of them may be mixed for use. Among them, water is preferably used.

[0216] The amount of the solvent used is usually 0.1 to 100 times by volume relative to compound (VI).

[0217] For example, when iodine is used, the reaction is preferably carried out by adding (preferably dropping) a mixture of compound (VI) and concentrated ammonia (and a solvent as necessary) to a mixture of iodine, potassium iodide and a solvent.

[0218] The reaction is usually carried out at -100 to 20° C., preferably -20 to 10° C. The reaction time also depends on the reaction temperature, but is usually about 30 minutes to about 24 hours, preferably about 1 hour to about 12 hours.

[0219] Completion of the reaction can be confirmed by thin layer chromatography, liquid chromatography or the like.

[0220] After the reaction is completed, compound (V) can be isolated and / or purified from the reaction mixture by conventional methods by separation means such as concentration, crystallization, recrystallization, distillation, solvent extraction, fractionation, chromatography, etc.

[0221] Through this reaction, an iodine atom or a bromine atom is introduced at the 3-position of the benzene ring in the case of α-methyltyrosine (the hydroxyl group is at the 4-position of the benzene ring), and at the 4-position or 6-position of the benzene ring in the case of α-methyl-m-tyrosine (the hydroxyl group is at the 3-position of the benzene ring).

[0222] Step 3 is to protect the amino and carboxyl groups of compound (V) (introducing P 1 and P 2 ), and R 2 A step of introducing a hydroxyl group to obtain a compound represented by formula (IV) or a salt thereof (hereinafter also referred to as compound (IV)).

[0223] R 2 The introduction of can be carried out by reacting with a corresponding iodide or bromide in the presence of a base.

[0224] The amount of the corresponding iodide or bromide used is usually 1 to 5 moles, preferably 1 to 2 moles, relative to 1 mole of the substrate.

[0225] Examples of the base include potassium carbonate, sodium carbonate, sodium hydrogen carbonate, and cesium carbonate.

[0226] The amount of the base used is usually 1 to 5 moles, preferably 1 to 2 moles, relative to 1 mole of the substrate.

[0227] The reaction is usually carried out in a solvent. As the solvent used in the present invention, there is no particular limitation as long as it has no adverse effect on the reaction, and examples thereof include amide solvents such as dimethylformamide, dimethylacetamide, and N-methylpyrrolidone; ether solvents such as tetrahydrofuran and 1,4-dioxane, and the like, and two or more of them may be mixed for use. Among them, dimethylformamide is preferably used.

[0228] The amount of the solvent used is usually 0.1 to 100 times the volume of the substrate.

[0229] The reaction is usually carried out at 0 to 100° C., preferably at room temperature to 50° C. The reaction time also depends on the reaction temperature, but is usually about 30 minutes to about 24 hours, preferably about 1 hour to about 17 hours.

[0230] Completion of the reaction can be confirmed by thin layer chromatography, liquid chromatography or the like.

[0231] After the reaction is completed, the target product can be separated and / or purified from the reaction mixture by conventional methods such as concentration, crystallization, recrystallization, distillation, solvent extraction, fractionation, chromatography, etc.

[0232] Carboxyl protecting group (P 1 ) and amino protecting group (P 2 ) Select a method that can be deprotected under mild conditions in step 5 to prevent the introduction of R 2 A protecting group that is detachable from a boron group or its ester group.

[0233] In addition, since the boronic acid compound (II) can be obtained as a crystal, it is easy to purify and a product of higher purity can be obtained, it is particularly preferably obtained as a boronic acid pinacol ester (Y is 4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl). Therefore, it is best to select a protecting group that can be deprotected in step 5 under the condition that Y is not hydrolyzed.

[0234] A preferred amino protecting group is tert-butoxycarbonyloxy.

[0235] Preferred carboxyl protecting groups are benzyl or C 1-2 The alkyl group (methyl group, ethyl group) is particularly preferably a benzyl group.

[0236] The introduction of each protecting group can be carried out according to a method known per se.

[0237] R 2 The introduction of the hydroxyl group is carried out by reaction with the corresponding iodide or bromide as described above, so the R 2 is also introduced into the amino group. 2 Previously, the amino protecting group (P 2 ). In addition, when importing the above R 2 When R is introduced into the carboxyl group 2 , but you can also remove the R if necessary 2 After that, the desired carboxyl protecting group (P 1 ).

[0238] As a preferred combination, R 2 C 1-6 Alkyl (preferably methyl, ethyl, propyl) or C 1-6 A haloalkyl group (preferably trifluoromethyl or 2-fluoroethyl), a carboxyl protecting group (P 1 ) is benzyl, the amino protecting group (P 2 ) is a combination of tert-butoxycarbonyloxy, R 2 C 7-14 Arylalkyl (preferably benzyl), carboxyl protecting group (P 1 ) is C 1-2 Alkyl (methyl, ethyl), amino protecting group (P2 ) is a combination of tert-butoxycarbonyloxy.

[0239] After the reaction is completed, compound (IV) can be isolated and / or purified from the reaction mixture by conventional methods by concentration, crystallization, recrystallization, distillation, solvent extraction, fractionation, chromatography and the like.

[0240] Among the compounds (IV), the compound represented by the following formula (IV') is a novel compound.

[0241]

[0242] [Where R 2a Represents C 1-3 Alkyl, P 1a Represents benzyl or C 1-3 alkyl.]

[0243] Step 4 is a step of reacting compound (IV) with a boronic acid introducing reagent in the presence of a palladium catalyst and a base to obtain a boronic acid compound (III).

[0244] Examples of the boronic acid introduction reagent include 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-1,3,2-dioxaborolane (also known as bi-boronic acid pinacol ester), 4,4,5,5-trimethyl-1,3,2-dioxaborolane, tetrahydroxydiborane, etc. Among them, bi-boronic acid pinacol ester is preferably used. These boronic acid introduction reagents can be commercially available.

[0245] The amount of the boronic acid-introducing agent used is generally 1 to 10 mol, preferably 1 to 3 mol, based on 1 mol of compound (IV).

[0246] Examples of the palladium catalyst include [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (PdCl2(dppf)) or its dichloromethane adduct, palladium acetate, and tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4). Among them, PdCl2(dppf) is preferably used.

[0247] The amount of the palladium catalyst to be used is generally 0.001 to 1 mol, preferably 0.01 to 0.2 mol, based on 1 mol of compound (IV).

[0248] Examples of the base include alkali metal acetates such as potassium acetate and sodium acetate. Among them, alkali metal acetates are preferably used, and potassium acetate is particularly preferably used.

[0249] The amount of the base to be used is generally 0.5 to 10 mol, preferably 1 to 5 mol, based on 1 mol of compound (IV).

[0250] The reaction is usually carried out in a solvent. As the solvent used in the present invention, there is no particular limitation as long as it has no adverse effect on the reaction, and examples thereof include sulfoxide solvents such as dimethyl sulfoxide; amide solvents such as dimethylformamide, dimethylacetamide, and N-methylpyrrolidone; and ether solvents such as tetrahydrofuran and 1,4-dioxane, and two or more of them may be mixed for use. Among them, sulfoxide solvents and amide solvents are preferably used, and dimethyl sulfoxide is particularly preferably used.

[0251] The amount of the solvent used is usually 0.1 to 100 times the volume of Compound (IV).

[0252] The reaction is preferably carried out, for example, by adding (preferably dropping) a boronic acid-introducing reagent to a mixture of compound (IV), a palladium catalyst, a base and a solvent.

[0253] The reaction is usually carried out at 0 to 200° C., preferably at room temperature to 120° C. The reaction time also depends on the reaction temperature, but is usually about 10 minutes to about 48 hours, preferably about 1 hour to about 12 hours.

[0254] Completion of the reaction can be confirmed by thin layer chromatography, liquid chromatography or the like.

[0255] After the reaction is completed, the boronic acid compound (III) can be separated and / or purified from the reaction mixture by conventional methods by concentration, crystallization, recrystallization, distillation, solvent extraction, fractionation, chromatography and the like.

[0256] Boronic acid compound (III) is a new type of compound.

[0257] The boronic acid compound (III) may be any of the boronic acid compound (IIIa), compound (IIIb) and racemic form represented by the following formula.

[0258]

[0259] [The symbols in the formula have the same meanings as above.]

[0260] In one embodiment, the boronic acid compound (III) is preferably a boronic acid compound (III a).

[0261] Step 5 is to remove the carboxyl protecting group (P 1 ) and amino protecting group (P 2 ) is a step of obtaining a boronic acid compound (II).

[0262] The removal of each protecting group is carried out according to a method known per se. 2 C 1-6 Alkyl (preferably methyl, ethyl, propyl) or C 1-6A haloalkyl group (preferably trifluoromethyl or 2-fluoroethyl) and a carboxyl protecting group (P 1 ) is benzyl, P 1 The removal is carried out by catalytic hydrogenation.

[0263] In this case, when Y is 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl, Y is not hydrolyzed, and thus the boronic acid compound (II) can be obtained as boronic acid pinacol ester.

[0264] In R 2 C 7-14 Arylalkyl (preferably benzyl) and carboxyl protecting group (P 1 ) is C 1-2 In the case of alkyl (methyl, ethyl), P 1 The removal is carried out by treating with an alkali such as lithium hydroxide or sodium hydroxide.

[0265] In the case of amino protecting groups (P 2 When ) is tert-butyloxycarbonyl, the removal is carried out by treating with an acid such as trifluoroacetic acid or hydrogen chloride.

[0266] The removal of the above protecting groups is performed under mild conditions and R 2 and the detachment of the boron group (-B(OH)2) or its ester group.

[0267] After the reaction is completed, the boronic acid compound (II) can be separated and / or purified from the reaction mixture by conventional methods by concentration, crystallization, recrystallization, distillation, solvent extraction, fractionation, chromatography and the like.

[0268] The reaction conditions such as solvent and reaction temperature in each step of the production method of the present invention described above are described in detail as representative examples in the embodiments described below, but are not necessarily limited to these. As long as a technician in this technical field can select them appropriately based on general knowledge in organic synthesis.

[0269] The radiolabeled compound (I) thus produced specifically binds to LAT1 whose expression is enhanced in tumor or cancer cells, and then is taken up into the cells and stably accumulated.

[0270] Thus, since the radiolabeled compound (I) targets tumor or cancer cells expressing LAT1, the radiolabeled compound (I) containing a therapeutically effective radionuclide can be used to treat tumors or cancers expressing LAT1. Examples of therapeutically effective radionuclides include 211 At 210 At 131 I. 125 I and 77 Br.

[0271] Examples of tumors or cancers expressing LAT1 include pancreatic cancer, leukemia, melanoma, colorectal cancer, lung cancer, prostate cancer, stomach cancer, breast cancer, kidney cancer, laryngeal cancer, esophageal cancer, liver cancer, lymphoma, myeloma, head and neck cancer, ovarian cancer, bladder cancer, childhood cancer, childhood leukemia, brain tumors (including primary brain tumors and metastatic brain tumors), osteosarcoma, soft tissue sarcoma, bone and soft tissue tumors, and the like.

[0272] In addition, the radiolabeled compound (I) has a higher retention rate in tumors or cancer sites and has a clearance rate that does not cause side effects, and can therefore exert a better antitumor effect in the living body.

[0273] Furthermore, the radiolabeled compound (I) can treat tumors or cancers expressing LAT1 while showing almost no side effects.

[0274] Furthermore, since the radiolabeled compound (I) is stable in blood and urine, it is considered to be stable until it reaches the tumor and is excreted.

[0275] In addition, since the radiolabeled compound (I) can also exert excellent antitumor effects on tumors or cancers that have acquired anticancer drug resistance (for example, platinum preparations (such as cisplatin) resistance, nucleic acid synthesis inhibitors (such as gemcitabine) resistance), the necessity of conventional treatment strategies for tumors or cancers that have acquired anticancer drug resistance, such as increasing the drug dose, changing the drug, changing the treatment method, etc., may be reduced.

[0276] The dosage of the radiolabeled compound (I) for therapeutic purposes is usually determined by the radionuclide used, the patient's weight, age, sex, treatment site, etc. For example, in the case of human subjects, 211 The standard amount of At radioactively labeled compound (I) considered to be effective per administration is about 100 MBq to 900 MBq.

[0277] The radiolabeled compound (I) is usually mixed with a pharmaceutically acceptable carrier for use as a pharmaceutical composition. A pharmaceutically acceptable carrier refers to a biocompatible solution that takes into full consideration sterility, pH, isotonicity, stability, etc., and may include various solvents, diluents (including sterile saline, sodium chloride injection, Ringer's injection, glucose injection, glucose and sodium chloride injection, lactated Ringer's injection, and other aqueous buffers), dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, etc. Pharmaceutically acceptable carriers may also include stabilizers, preservatives, antioxidants, or other additives known to those skilled in the art.

[0278] The dosage form of the pharmaceutical composition is not particularly limited, and can be prepared as a pharmaceutical composition for oral administration in the form of granules, fine granules, powders, hard capsules, soft capsules, syrups, emulsions, suspensions or liquids, or a pharmaceutical composition for parenteral administration in the form of injections, drips, percutaneous absorption agents, transmucosal absorption agents, nasal drops, inhalants, suppositories, etc. for intravenous administration, intramuscular administration or subcutaneous administration. These preparations can be prepared according to conventional methods. Preferably, they are liquid preparations for oral administration or injection.

[0279] The liquid preparation is prepared by dissolving the radiolabeled compound (I) in water, but can also be dissolved in physiological saline or glucose solution as required, and a buffer or preservative can also be added in addition. In addition, as mentioned above, reducing agents such as ascorbic acid or ascorbic acid alkali metal salts or alkaline earth metal salts can also be contained. In particular, in order to manufacture injection, the effective ingredient can be dissolved in injection distilled water together with the isotonic agents of pH adjusting agents such as hydrochloric acid, sodium hydroxide, lactose, lactic acid, sodium, sodium monohydrogen phosphate, sodium dihydrogen phosphate, sodium chloride, glucose, etc. as required, and filled into ampoule after aseptic filtration, further, mannitol, dextrin, cyclodextrin, gelatin, etc. can also be added for vacuum freeze drying, as the injection dissolved when used. In addition, lecithin, polysorbate 80, polyoxyethylene hydrogenated castor oil, etc. can also be added to the effective ingredient, emulsified in water, and injection emulsion is made.

[0280] The half-life of the radionuclide contained in the radiolabeled compound (I) is 211 At is as short as 7.2 hours, 210 At is as short as 8.3 hours, 131 I is short for 8.04 days, 125 I is short at 59.4 days, 77 Br is as short as 57 hours, so it is desirable to prepare the pharmaceutical composition in a form containing the radiolabeled compound (I) in the amount required for administration prior to administration to a subject.

[0281] In addition, the boric acid compound (II) is a compound containing boron that targets tumor or cancer cells expressing LAT1 and can be used as a drug for boron neutron capture therapy (BNCT) combined with neutron irradiation. This therapy utilizes boron ( 10 B) It has a large capture cross-section for thermal neutrons. 10 B releases helium nuclei from nuclear reactions with thermal neutrons ( 4 He 2+ ) and lithium core ( 7 Li +) to destroy cancer cells. Both the helium nucleus and the lithium nucleus have high linear energy and a short range close to the size of cells. Therefore, by irradiating neutrons to the boric acid compound (II) taken into tumor or cancer cells, only the tumor or cancer cells can be effectively destroyed.

[0282] [Example]

[0283] The present invention is further described in detail by the following examples, but they are only examples and do not limit the present invention, and changes may be made without departing from the scope of the present invention.

[0284] In the following Examples, the radiochemical yield was calculated by the following formula.

[0285] Radiochemical yield (%) = (radioactivity of target compound in thin layer plate / total radioactivity in thin layer plate) × 100

[0286] Example 1

[0287] (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-methyl-α-methyltyrosine hydrochloride ( 6 )(Compound( 6 Synthesis of )Bpin-AMT-OMe)

[0288]

[0289] a)(S)-3-iodo-α-methyltyrosine( 2 )(Compound( 2 Synthesis of

[0290] 7.1 g of iodine (28.1 mmol) was added to a mixture of 5.3 g of potassium iodide (32.1 mmol) and 14 mL of water, and the mixture was stirred for 2 hours. 5.2 g of (S)-α-methyltyrosine ( 1 ) (26.8mmol), 62mL of concentrated ammonia water (28%) and 8mL of water were stirred at -7 to -5°C for 3 hours. 15mL of 15% sodium sulfite aqueous solution was added to the reaction solution, and after warming to room temperature, 6M hydrochloric acid was added under ice-cooling to adjust the pH to 6.5 to 7. After stirring under ice-cooling for 1 hour, the precipitated solid was filtered. After washing with cold water and acetone, it was dried under reduced pressure to obtain 7.4g of compound ( 2 ) (yield 86%).

[0291] 1H-NMR(300MHz,DMSO-d6+TFA,TMS):8.25(3H,br),7.51(1H,d,J=2.1Hz),7.03(1H,dd,J=2.1 ,8.1Hz),6.84(1H,d,J=8.1Hz),3.03(1H,d,J=14.1Hz),2.87(1H,d,J=14.1Hz),1.44(3H,s)

[0292] b) (S)-N-Boc-3-iodo-O-methyl-α-methyltyrosine methyl ester ( 3 )(Compound( 3 Synthesis of

[0293] In 4.3g of compound ( 2 )(13.4mmol), 13mL 1M sodium hydroxide aqueous solution and 20mL 1,4-dioxane mixed solution, add 8.8g di-tert-butyl dicarbonate (40.2mmol), and stir at 60℃ for 20 hours. After cooling the reaction solution to room temperature, concentrate the reaction solution under reduced pressure, add 30mL water and 60mL methyl tert-butyl ether-heptane (1:2) to the residue, and perform liquid separation extraction. Back-extract the organic layer with 20mL 5% sodium bicarbonate aqueous solution and 20mL water. Add 0.5M sodium bisulfate aqueous solution to the combined water layer to adjust the pH to 1-2, and extract with 60mL methyl tert-butyl ether-ethyl acetate (1:1). Wash the extract with water and saturated brine. Remove the solvent by distillation under reduced pressure to obtain 2.9g of light brown solid. The obtained 2.9 g of light brown solid was dissolved in 20 mL of N,N-dimethylformamide, and 2.1 g of potassium carbonate (15 mmol) and 2.1 g of iodomethane (15 mmol) were added, and stirred at room temperature for 17 hours. 60 mL of ethyl acetate was added, and the solvent was removed by vacuum distillation after washing with 60 mL of water and 30 mL of saturated brine. The residue was purified by silica gel column chromatography to obtain 2.6 g of a white amorphous compound ( 3 ) (yield 43%).

[0294] 1 H-NMR (300MHz, CDCl3, TMS): 7.50 (1H, d, J = 2.1Hz), 7.00 (1H, dd, J = 2.1, 8.4Hz), 6.71 (1H, d, J = 8.4Hz), 5.17 ( 1H,brs),3.85(3H,s),3.77(3H,s),3.33(1H,d,J=13.5Hz),3.10(1H,d,J=13.5Hz),1.55(3H,s),1.49(9H,s)

[0295] c) (S)-N-Boc-3-iodo-O-methyl-α-methyltyrosine benzyl ester (4 )(Compound( 4 Synthesis of

[0296] In the compound ( 3 ) 7mL of methanol (2.6g (5.8mmol) and 13mL of tetrahydrofuran) were added with 7.0mL (7.0mmol) of 1M sodium hydroxide aqueous solution, and stirred at room temperature for 4 hours and at 60°C for 19 hours. After cooling the reaction solution to room temperature, the reaction solution was concentrated. 30mL of water and 30mL of methyl tert-butyl ether were added to the residue for separation, and 0.5M sodium bisulfate aqueous solution was added to the water layer to adjust the pH to 2-3, and then extracted with 30mL of methyl tert-butyl ether. After washing the extract with water, the solvent was removed by vacuum distillation to obtain 2.3g of white amorphous. The obtained 2.3g of white amorphous (5.3mmol) was dissolved in 14mL of N,N-dimethylformamide, 880mg of potassium carbonate (6.3mmol) and 0.75mL of benzyl bromide (6.3mmol) were added, and stirred at 50°C for 4 hours. The mixture was cooled to room temperature, 30 mL of ethyl acetate was added, and the solvent was removed by distillation under reduced pressure after washing with water and saturated brine. The residue was purified by silica gel column chromatography to obtain 2.4 g of a white amorphous compound ( 4 ) (yield 79%).

[0297] 1 H-NMR (300MHz, DMSO-d6, TMS): 7.52-7.30 (7H, m), 7.00 (1H, dd, J = 2.1, 8.4Hz), 6.92 (1H, d, J = 8.4Hz), 5.12 (1H, d, J = 1 2.6Hz),5.03(1H,d,J=12.6Hz),3.79(3H,s),3.28(1H,d,J=15.0Hz),2.72(1H,d,J=15.0Hz),1.42(9H,s),1.16(3H,s)

[0298] d) (S)-N-Boc-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-O-methyl-α-methyltyrosine benzyl ester ( 5 )(Compound( 5 Synthesis of

[0299] Under nitrogen atmosphere, 2.4 g of compound ( 4)(4.6mmol), 0.29g [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (II) dichloromethane adduct (0.35mmol), 1.79g potassium acetate (18.3mmol) and 19mL dimethyl sulfoxide mixed solution, add 2.32g of boric acid pinacol ester (9.1mmol), after stirring at room temperature for 1 hour, stir at 55°C for 2 hours. The reaction solution was cooled to room temperature, 50mL of methyl tert-butyl ether and 50mL of water were added, and filtered through diatomaceous earth. After separation, the aqueous layer was extracted with 30mL of methyl tert-butyl ether, the organic layers were combined, washed with 30mL of water and 30mL of saturated brine, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography to obtain 1.6g of a white amorphous compound ( 5 ) (yield 67%).

[0300] 1 H-NMR (300MHz, CDCl3, TMS): 7.5-7.3 (6H, m), 7.06 (1H, dd, J = 2.1, 8.4Hz), 6.72 (1H, d, J = 8.4Hz), 5.21 (1H, d, J = 12.6Hz), 5 .13(1H,d,J=12.6Hz),3.80(3H,s),3.31(1H,d,J=13.5Hz),3.16(1H,d,J=13.5Hz),1.46(9H,s),1.32(12H,s),1.26(3H,s)

[0301] e) (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-O-methyl-α-methyltyrosine hydrochloride ( 6 )(Compound( 6 Synthesis of )Bpin-AMT-OMe)

[0302] 1.3 g of compound ( 5 )(2.5mmol), 0.33g 10% palladium-carbon (55% water) and 13mL tetrahydrofuran were stirred for 3 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth and the solvent was distilled off under reduced pressure to obtain 1.1g of white amorphous. 4mL of 4M hydrochloric acid-ethyl acetate solution was added to the obtained mixture of 1.1g amorphous (2.5mmol) and 4.0mL ethyl acetate, and stirred at room temperature for 18 hours. The precipitated solid was filtered and washed with ethyl acetate to obtain 0.51g of the compound as a white powder ( 6 ) (yield 55%).

[0303] 1H-NMR (300MHz, DMSO-d6, TMS): 8.35 (3H, br), 7.39 (1H, d, J = 2.4Hz), 7.28 (1H, dd, J = 2.4, 8.4Hz), 6.94 ( 1H,d,J=8.4Hz),3.72(3H,s),3.08(1H,d,J=13.8Hz),2.98(1H,d,J=13.8Hz),1.46(3H,s),1.27(12H,s)

[0304] HPLC analysis conditions

[0305] Sample solution: 1 mg of compound ( 6 ) was dissolved in 1 mL of 67% acetonitrile water to prepare a sample solution.

[0306] Detector: UV 275nm

[0307] Chromatographic column: YMC-Pack Pro C18 RS ( 5μm,YMC)

[0308] Column temperature: 30°C

[0309] Mobile phase: H2O (0.1% TFA) / MeCN (0.1% TFA)

[0310] Mode: Gradient

[0311] 0min H2O(0.1%TFA) / MeCN(0.1%TFA)=95 / 5

[0312] 30min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0313] 40min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0314] Flow rate: 1mL / min

[0315] Injection volume: 5μL

[0316] Analysis time: 40 minutes

[0317] Holding time: Compound ( 6 )8.1 minutes

[0318] HPLC chart Figure 1 shown.

[0319] Reference Example 1

[0320] 211 Preparation of aqueous solution of At

[0321] According to the method described in Reference Example 1 of Patent Document 2, 211 At aqueous solution. That is, 211 At is a method of irradiating bismuth with helium particles (28 MeV) accelerated by a cyclotron. 209 Bi(α, 2n) 211 After irradiation, the target substance 209 Bi is heated and melted, 211 At evaporates and is collected in a cold trap and dissolved with a small amount of water.

[0322] Example 2

[0323] (S)-3-Astatine( 211 At)-O-methyl-α-methyltyrosine ( 211 Synthesis of At-AAMT-OMe

[0324]

[0325] Sodium bicarbonate (Meylon) (7%) was used to dissolve the compound ( 6 ) (Bpin-AMT-OMe), prepare an aqueous solution (1 mg / mL), and place the aqueous solution (0.1 mL) in a 1.5 mL polypropylene (PP) tube. Then, add the 211 At aqueous solution (100 μL, 10 MBq, measured with a Curie meter (IGC-7, Hitachi)) and 0.1 M KI aqueous solution (20 μL) were added, followed by sterile distilled water to make the total volume 200 μL. The mixture was reacted at 50°C for 45 minutes to obtain (S)-3-astatine ( 211 At)-O-methyl-α-methyltyrosine ( 211 At-AAMT-OMe) crude product solution.

[0326] The crude product solution (1 μL) was analyzed by thin layer chromatography (TLC). The sample was applied to a thin layer plate (silica gel 60F 254 ) was used as the developing solvent using a mixture of acetonitrile and water (2 / 1). After the thin layer plate was exposed on an imaging plate (BAS IP MS2025E, GE Healthcare) for about 15 minutes, the plate was analyzed by a bio-image analyzer (Tyhoon TM FLA-7000, Cytiva) was used for analysis. The results are shown in Figure 2 As shown. The crude product solution was detected at Rf0.80 211 At-AAMT-OMe, radiochemical yield (RCY) was 81.3%.

[0327] The crude product solution was injected into an HLB column (Oasis, WAT-186005125) equilibrated with sterile distilled water (1 mL), the target product was captured in the cartridge, and impurities were eluted by passing sterile distilled water. Then, a 30% ethanol solution (500 μL) was passed through the cartridge, and the target product was eluted into an aqueous sodium ascorbate solution (500 μL, final concentration 1%) to obtain a purified product. The radiochemical yield of the purified product was 81%, and the radiochemical purity was 98%.

[0328] In addition, the samples were analyzed by thin layer chromatography (TLC) in the same manner as above before purification, immediately after production, 3 hours after production, and 24 hours after production. 211 At-AAMT-OMe. The result is as follows Figure 3 As shown. Figure 3 It can be seen that even after 24 hours of purification, no 211 Decomposition of At-AAMT-OMe.

[0329] Example 3

[0330] (R)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-methyl-α-methyltyrosine hydrochloride ( 12 )(Compound( 12 Synthesis of (R)-Bpin-AMT-OMe)

[0331]

[0332] a)(R)-3-iodo-α-methyltyrosine ( 8 )(Compound( 8 Synthesis of

[0333] 6.8 g of iodine (26.9 mmol) was added to a mixture of 5.1 g of potassium iodide (30.7 mmol) and 14 mL of water, and the mixture was stirred for 2 hours. 5.0 g of (R)-α-methyltyrosine ( 7 ) (25.6mmol), 59mL of concentrated ammonia water (28%) and 8mL of water were stirred at -7 to -5°C for 3 hours. 15mL of 15% sodium sulfite aqueous solution was added to the reaction solution, and after warming to room temperature, 6M hydrochloric acid was added under ice-cooling to adjust the pH to 6.5 to 7. After stirring for 1 hour under ice-cooling, the precipitated solid was filtered. After washing with cold water and acetone, it was dried under reduced pressure to obtain 7.1g of compound ( 8 ) (yield 86%).

[0334] 1H-NMR (400MHz, DMSO-d6+TFA, TMS): 8.25 (3H, br), 7.51 (1H, d, J = 2.0Hz), 7.03 (1H, dd, J = 2.0 ,8.4Hz),6.84(1H,d,J=8.4Hz),3.03(1H,d,J=14.1Hz),2.87(1H,d,J=14.1Hz),1.44(3H,s)

[0335] b) (R)-N-Boc-3-iodo-O-methyl-α-methyltyrosine methyl ester ( 9 )(Compound( 9 Synthesis of

[0336] In the compound ( 8 )4.3g (13.4mmol), 13mL of 1M sodium hydroxide aqueous solution and 20mL of 1,4-dioxane were added with 8.8g (40.2mmol) of di-tert-butyl dicarbonate, and stirred at 60°C for 20 hours. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure, and 30mL of water and 60mL of methyl tert-butyl ether-heptane (1:2) were added to the residue for liquid separation and extraction. The organic layer was washed with 20mL of 5% sodium bicarbonate aqueous solution and 20mL of water. 0.5M sodium bisulfate aqueous solution was added to the combined washing water layer to adjust the pH to 1-2, and extracted with 60mL of methyl tert-butyl ether-ethyl acetate (1:1). The extract was washed with water and saturated brine. The solvent was removed by distillation under reduced pressure to obtain 4.2g of light brown solid. The obtained 4.2 g of light brown solid was dissolved in 20 mL of N,N-dimethylformamide, and 3.0 g of potassium carbonate (21.9 mmol) and 3.1 g (21.9 mmol) of methyl iodide were added, and the mixture was stirred at room temperature for 17 hours. 60 mL of ethyl acetate was added, and the solvent was removed by distillation under reduced pressure after washing with 60 mL of water and 30 mL of saturated brine. The residue was purified by silica gel column chromatography to obtain 3.2 g of a white amorphous compound ( 9 ) (yield 53%).

[0337] 1 H-NMR (400MHz, CDCl3, TMS): 7.50 (1H, d, J = 2.0Hz), 7.00 (1H, dd, J = 2.0, 8.4Hz), 6.71 (1H, d, J = 8.4Hz), 5.17 ( 1H,brs),3.85(3H,s),3.77(3H,s),3.33(1H,d,J=13.6Hz),3.10(1H,d,J=13.6Hz),1.55(3H,s),1.49(9H,s)

[0338] c) (R)-N-Boc-3-iodo-O-methyl-α-methyltyrosine benzyl ester ( 10 )(Compound( 10 Synthesis of

[0339] In the compound ( 9 )3.2g (7.1mmol) of methanol 13mL, tetrahydrofuran 15mL mixed solution, add 2M sodium hydroxide aqueous solution 12.8mL (25.6mmol), stir at 60℃ for 24 hours. After cooling the reaction solution to room temperature, concentrate the reaction solution. Add 30mL of water and 30mL of methyl tert-butyl ether to the residue for separation, add 0.5M sodium bisulfate aqueous solution to the water layer, adjust the pH to 2-3, and extract with 30mL of methyl tert-butyl ether. After washing the extract with water, remove the solvent by vacuum distillation to obtain 3.0g of white amorphous. Dissolve the obtained 3.0g of white amorphous (6.9mmol) in 20mL of N,N-dimethylformamide, add 1.2g of potassium carbonate (8.5mmol) and 1.0mL of benzyl bromide (8.5mmol), and stir at 50℃ for 4 hours. The mixture was cooled to room temperature, 30 mL of ethyl acetate was added, and the solvent was removed by distillation under reduced pressure after washing with 30 mL of water and 30 mL of saturated brine. The residue was purified by silica gel column chromatography to obtain 3.5 g of a white amorphous compound ( 10 ) (yield 94%).

[0340] 1 H-NMR (400MHz, DMSO-d6, TMS): 7.52-7.30 (7H, m), 7.00 (1H, dd, J = 2.0, 8.4Hz), 6.92 (1H, d, J = 8.4Hz), 5.12 (1H, d, J = 1 2.4Hz),5.03(1H,d,J=12.4Hz),3.79(3H,s),3.28(1H,d,J=13.6Hz),2.72(1H,d,J=13.6Hz),1.42(9H,s),1.16(3H,s)

[0341] d) (R)-N-Boc-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-methyl-α-methyltyrosine benzyl ester ( 11) (Compound( 11 Synthesis of

[0342] Under nitrogen atmosphere, 3.5 g of compound ( 10)(6.6mmol), 0.41g [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (II) dichloromethane adduct (0.51mmol), 2.62g potassium acetate (26.7mmol) and 26mL dimethyl sulfoxide mixed solution, add 3.38g of boric acid pinacol ester (13.3mmol), after stirring at room temperature for 1 hour, stir at 55°C for 2 hours. The reaction solution was cooled to room temperature, 50mL of methyl tert-butyl ether and 50mL of water were added, and filtered through diatomaceous earth. After separation, the aqueous layer was extracted with 30mL of methyl tert-butyl ether, the organic layers were combined, washed with 30mL of water and 30mL of saturated brine, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography to obtain 1.7g of a white amorphous compound ( 11 ) (yield 72%).

[0343] 1 H-NMR (400MHz, CDCl3, TMS): 7.5-7.3 (6H, m), 7.06 (1H, dd, J = 2.0, 8.4Hz), 6.72 (1H, d, J = 8.4Hz), 5.21 (1H, d, J = 12.4Hz), 5 .13(1H,d,J=12.4Hz),3.80(3H,s),3.31(1H,d,J=13.6Hz),3.16(1H,d,J=13.6Hz),1.46(9H,s),1.32(12H,s),1.26(3H,s)

[0344] e) (R)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-methyl-α-methyltyrosine hydrochloride ( 12 )(Compound( 12 Synthesis of (R)-Bpin-AMT-OMe)

[0345] 1.7 g of compound ( 11 )(3.2mmol), 0.42g 10% palladium-carbon (55% water) and 17mL tetrahydrofuran were stirred in a hydrogen atmosphere for 3 hours. The reaction solution was filtered through diatomaceous earth and the solvent was distilled off under reduced pressure to obtain 1.4g of white amorphous. 5mL of 4M hydrochloric acid-ethyl acetate solution was added to the obtained mixture of 1.4g amorphous (3.2mmol) and 5.0mL ethyl acetate, and stirred at room temperature for 17 hours. The precipitated solid was filtered and washed with ethyl acetate to obtain 0.66g of white powder. 15.0mL ethyl acetate was added and suspended for washing at room temperature for 1 hour. The solid was filtered and washed with ethyl acetate to obtain 0.53g of the compound as a white powder ( 12 ) (yield 44%).

[0346] 1H-NMR (400MHz, DMSO-d6, TMS): 8.35 (3H, br), 7.39 (1H, d, J = 2.4Hz), 7.28 (1H, dd, J = 2.4, 8.4Hz), 6.94 ( 1H,d,J=8.4Hz),3.72(3H,s),3.08(1H,d,J=14.0Hz),2.98(1H,d,J=14.0Hz),1.46(3H,s),1.27(12H,s)

[0347] HPLC analysis conditions

[0348] Sample solution: 1 mg of compound ( 12 ) was dissolved in 1 mL of 67% acetonitrile water to prepare a sample solution.

[0349] Detector: UV 275nm

[0350] Chromatographic column: YMC-Pack Pro C18 RS ( 5μm,YMC)

[0351] Column temperature: 30°C

[0352] Mobile phase: H2O (0.1% TFA) / MeCN (0.1% TFA)

[0353] Mode: Gradient

[0354] 0min H2O(0.1%TFA) / MeCN(0.1%TFA)=95 / 5

[0355] 30min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0356] 40min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0357] Flow rate: 1mL / min

[0358] Injection volume: 5μL

[0359] Analysis time: 40 minutes

[0360] Holding time: Compound ( 12 )7.3 minutes

[0361] HPLC chart Figure 4 shown.

[0362] Example 4

[0363] (R)-3-Astatine( 211At)-O-methyl-α-methyltyrosine ((R)- 211 Synthesis of At-AAMT-OMe

[0364]

[0365] Sodium bicarbonate (7%) was used to dissolve the compound ( 12 ) ((R)-Bpin-AMT-OMe), prepare an aqueous solution (1 mg / mL), and place the aqueous solution (0.1 mL) in a 1.5 mL polypropylene (PP) tube. Then, add the 211 At aqueous solution (0.1 mL, 5 MBq, measured with a Curie meter (IGC-7, Hitachi)) and 0.1 M KI aqueous solution (20 μL) were added, followed by sterile distilled water to make the total volume 200 μL. The mixture was reacted at 50°C for 45 minutes to obtain (R)-3-astatine ( 211 At)-O-methyl-α-methyltyrosine ((R)- 211 At-AAMT-OMe) crude product solution.

[0366] The crude product solution (1 μL) was analyzed by thin layer chromatography (TLC). The sample was applied to a thin layer plate (silica gel 60F 254 ) was used as the developing solvent using a mixture of acetonitrile and water (2 / 1). After the thin layer plate was exposed on an imaging plate (BAS IP MS2025E, GE Healthcare) for about 15 minutes, the plate was analyzed by a bio-image analyzer (Tyhoon TM FLA-7000, Cytiva) was used for analysis. (R)- 211 At-AAMT-OMe, radiochemical yield (RCY) was 79.6%.

[0367] The crude product solution was injected into an HLB column (Oasis, WAT-186005125) equilibrated with sterile distilled water (1 mL), the target product was captured in the cartridge, and the sterile distilled water was passed to elute the impurities. Next, a 30% ethanol solution (500 μL) was passed through the cartridge to elute the target product into an aqueous sodium ascorbate solution (500 μL, final concentration 1%) to obtain a purified product. The purified product was analyzed by thin layer chromatography (TLC) in the same manner as above. The results are as follows: Figure 5 The radiochemical yield of the purified product was 79.6% and the radiochemical purity was 99.9%.

[0368] In addition, 24 hours after the production, (R)- 211 At-AAMT-OMe. The result is as follows Figure 6 As shown. Figure 6 It can be seen that even after 24 hours of purification, no (R)- 211 Decomposition of At-AAMT-OMe.

[0369] Example 5

[0370] (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-ethyl-α-methyltyrosine hydrochloride ( 18 )(Compound( 18 Synthesis of )Bpin-AMT-OEt)

[0371]

[0372] a)(S)-3-iodo-α-methyltyrosine( 14 )(Compound( 14 Synthesis of

[0373] 7.1 g of iodine (28.1 mmol) was added to a mixture of 5.3 g of potassium iodide (32.1 mmol) and 14 mL of water, and the mixture was stirred for 2 hours. 5.0 g of (S)-α-methyltyrosine ( 13 ) (25.6mmol), 62mL of concentrated ammonia water (28%) and 8mL of water were stirred at -7 to -5°C for 3 hours. 15mL of 15% sodium sulfite aqueous solution was added to the reaction solution, and after warming to room temperature, 6M hydrochloric acid was added under ice-cooling to adjust the pH to 6.5 to 7. After stirring under ice-cooling for 1 hour, the precipitated solid was filtered. After washing with cold water and acetone, it was dried under reduced pressure to obtain 8.4g of compound ( 14 ) (yield 98%).

[0374] 1 H-NMR (400MHz, DMSO-d6+TFA, TMS): 8.25 (3H, br), 7.51 (1H, d, J = 2.0Hz), 7.03 (1H, dd, J = 2.0 ,8.4Hz),6.84(1H,d,J=8.4Hz),3.03(1H,d,J=14.1Hz),2.87(1H,d,J=14.1Hz),1.44(3H,s)

[0375] b) (S)-N-Boc-3-iodo-O-ethyl-α-methyltyrosine ethyl ester ( 15 )(Compound( 15 Synthesis of

[0376] In 4.3g of compound ( 14)(13.4mmol), 31mL 1M sodium hydroxide aqueous solution and 50mL 1,4-dioxane, 25mL water mixed solution, add 3.3g di-tert-butyl dicarbonate (14.7mmol), stir at 60℃ for 22 hours. After cooling the reaction solution to room temperature, concentrate the reaction solution under reduced pressure, add 30mL water and 60mL methyl tert-butyl ether-heptane (1:2) to the residue, and perform liquid separation extraction. Wash the organic layer with 20mL 5% sodium bicarbonate aqueous solution and 20mL water. Add 0.5M sodium bisulfate aqueous solution to the combined washing water layer to adjust the pH to 1-2, and extract with 60mL methyl tert-butyl ether-ethyl acetate (1:1). Wash the extract with water and saturated brine. Remove the solvent by distillation under reduced pressure to obtain 3.1g of light brown solid. The obtained 3.1 g of light brown solid was dissolved in 20 mL of N,N-dimethylformamide, and 2.1 g of potassium carbonate (15 mmol) and 2.4 g of iodoethane (15 mmol) were added, and stirred at room temperature for 16 hours. 60 mL of ethyl acetate was added, and the solvent was removed by vacuum distillation after washing with 60 mL of water and 30 mL of saturated brine. The residue was purified by silica gel column chromatography to obtain 3.0 g of a white amorphous compound ( 15 ) (yield 47%).

[0377] 1 H-NMR (400MHz, CDCl3, TMS): 7.50 (1H, d, J = 2.0Hz), 6.99 (1H, dd, J = 2.0, 8.4Hz), 6.68 (1H, d, J = 8.4Hz), 5.18 (1H, brs), 4.21 (2H, q, J = 6.8Hz), 4. 06(2H,q,J=6.8Hz),3.33(1H,d,J=13.2Hz),3.10(1H,d,J=13.2Hz),1.5 4-1.57(3H,m),1.49(9H,s),1.46(3H,t,J=6.8Hz),1.32(3H,t,J=6.8Hz)

[0378] c) (S)-N-Boc-3-iodo-O-ethyl-α-methyltyrosine benzyl ester ( 16 )(Compound( 16 Synthesis of

[0379] In the compound ( 15)3.0g (6.3mmol) of methanol 15mL, tetrahydrofuran 15mL, 2M sodium hydroxide aqueous solution 11.3mL (22.6mmol) was added to the mixture, and stirred at 60°C for 17 hours. After the reaction solution was cooled to room temperature, the reaction solution was concentrated. 30mL of water and 30mL of methyl tert-butyl ether were added to the residue for separation, 0.5M sodium bisulfate aqueous solution was added to the water layer, and after the pH was adjusted to 2-3, it was extracted with 30mL of methyl tert-butyl ether. After washing the extract with water, the solvent was removed by vacuum distillation to obtain 2.9g of white amorphous. The obtained 2.9g of white amorphous (6.3mmol) was dissolved in 17mL of N,N-dimethylformamide, 1.0g of potassium carbonate (7.6mmol) and 0.87mL of benzyl bromide (7.6mmol) were added, and stirred at 50°C for 3 hours. The mixture was cooled to room temperature, 30 mL of ethyl acetate was added, and the solvent was removed by distillation under reduced pressure after washing with water and saturated brine. The residue was purified by silica gel column chromatography to obtain 3.2 g of a white amorphous compound ( 16 ) (yield 94%).

[0380] 1 H-NMR (400MHz, DMSO-d6, TMS): 7.52-7.30 (6H, m), 7.09 (1H, s), 7.07 (1H, dd, J = 2.0, 8.4Hz), 6.89 (1H, d, J = 8.4Hz), 5.07 (1H, d, J = 12.4Hz), 5 .04(1H,d,J=12.4Hz), 4.05(2H,q,J=6.8Hz), 3.28(1H,d,J=13.6Hz), 2.82(1H,d,J=13.6Hz), 1.41(9H,s), 1.34(3H,t,J=6.8Hz), 1.16(3H,s)

[0381] d) (S)-N-Boc-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-ethyl-α-methyltyrosine benzyl ester ( 17 )(Compound( 17 Synthesis of

[0382] Under nitrogen atmosphere, 3.2 g of compound ( 16)(5.9mmol), 0.37g [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (II) dichloromethane adduct (0.45mmol), 2.3g potassium acetate (23.7mmol) and 27mL dimethyl sulfoxide were added with 3.0g of diboric acid pinacol ester (11.8mmol), stirred at room temperature for 1 hour, and then stirred at 55°C for 2 hours. The reaction solution was cooled to room temperature, 50mL of methyl tert-butyl ether and 50mL of water were added, and diatomaceous earth was filtered. After separation, the aqueous layer was extracted with 30mL of methyl tert-butyl ether, the organic layers were combined, washed with 30mL of water and 30mL of saturated brine, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography to obtain 1.8g of a white amorphous compound ( 17 ) (yield 57%).

[0383] 1 H-NMR (400MHz, CDCl3, TMS): 7.42-7.30 (6H, m), 7.02 (1H, dd, J = 2.4, 8.4Hz), 6.70 (1H, d, J = 8.4Hz), 5.19 (1H, d, J = 12.4Hz), 5. 12(1H,d,J=12.4Hz),3.99(2H,q,J=6.8Hz),3.34-3.12(2H,m),1.45(9H,s),1.39(3H,t,J=6.8Hz),1.32(12H,s),1.26(3H,s)

[0384] e) (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-ethyl-α-methyltyrosine hydrochloride ( 18 )(Compound( 18 Synthesis of )Bpin-AMT-OEt)

[0385] 1.8 g of compound ( 17 )(3.3mmol), 0.45g 10% palladium-carbon (55% water) and 18mL tetrahydrofuran were stirred for 3 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth and the solvent was distilled off under reduced pressure to obtain 1.4g of white amorphous. 4.2mL of 4M hydrochloric acid-ethyl acetate solution was added to a mixture of 1.4g amorphous (3.1mmol) and 4.2mL ethyl acetate, and stirred at room temperature for 15 hours. The precipitated solid was filtered and washed with ethyl acetate to obtain 0.67g of white powder. 15.0mL ethyl acetate was added and suspended and washed at room temperature for 1 hour. The solid was filtered and washed with ethyl acetate to obtain 0.52g of the compound as a white powder ( 18 ) (yield 41%).

[0386] 1H-NMR (400MHz, DMSO-d6, TMS): 8.31 (3H, br), 7.31 (1H, d, J = 2.4Hz), 7.24 (1H, dd, J = 2.4, 8.4Hz), 6.91 (1H, d, J = 8.4Hz), 3.98(2H,q,J=6.8Hz), 3.07(1H,d,J=14.0Hz), 2.97(1H,d,J=14.0Hz), 1.45(3H,s), 1.29(3H,t,J=6.8Hz), 1.27(12H,s)

[0387] HPLC analysis conditions

[0388] Sample solution: 1 mg of compound ( 18 ) was dissolved in 1 mL of 67% acetonitrile water to prepare a sample solution.

[0389] Detector: UV 275nm

[0390] Chromatographic column: YMC-Pack Pro C18 RS ( 5μm,YMC)

[0391] Column temperature: 30°C

[0392] Mobile phase: H2O (0.1% TFA) / MeCN (0.1% TFA)

[0393] Mode: Gradient

[0394] 0min H2O(0.1%TFA) / MeCN(0.1%TFA)=95 / 5

[0395] 30min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0396] 40min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0397] Flow rate: 1mL / min

[0398] Injection volume: 5μL

[0399] Analysis time: 40 minutes

[0400] Holding time: Compound ( 18 ) 9.4 minutes

[0401] HPLC chart Figure 7 shown.

[0402] Example 6

[0403] (S)-3-Astatine( 211 At)-O-ethyl-α-methyltyrosine ( 211 Synthesis of At-AAMT-OEt)

[0404]

[0405] Sodium bicarbonate (7%) was used to dissolve the compound ( 18 ) (Bpin-AMT-OEt), prepare an aqueous solution (1 mg / mL), and put the aqueous solution (0.1 mL) into a 1.5 mL polypropylene (PP) tube. Then, add the 211 At aqueous solution (100 μL, 5 MBq, measured with a Curie meter (IGC-7, Hitachi)) and 0.1 M KI aqueous solution (20 μL) were added, followed by sterile distilled water to make the total volume 200 μL. The mixture was reacted at 50°C for 45 minutes to obtain (S)-3-astatine ( 211 At)-O-ethyl-α-methyltyrosine ( 211 At-AAMT-OEt) crude product solution.

[0406] The crude product solution (1 μL) was analyzed by thin layer chromatography (TLC). The sample was applied to a thin layer plate (silica gel 60F 254 ) was used as the developing solvent using a mixture of acetonitrile and water (2 / 1). After the thin layer plate was exposed on an imaging plate (BAS IP MS2025E, GE Healthcare) for about 15 minutes, the plate was analyzed by a bio-image analyzer (Tyhoon TM FLA-7000, Cytiva) was used for analysis. 211 At-AAMT-OEt, radiochemical yield (RCY) was 85.5%.

[0407] The crude product solution was injected into an HLB column (Oasis, WAT-186005125) equilibrated with sterile distilled water (1 mL), the target product was captured in the cartridge, and the impurities were eluted by passing the sterile distilled water. Next, a 30% ethanol solution (500 μL) was passed through the cartridge, and the target product was eluted into an aqueous sodium ascorbate solution (500 μL, final concentration 1%) to obtain a purified product. The purified product was analyzed by thin layer chromatography (TLC) in the same manner as above. The results are as follows: Figure 8 The radiochemical yield of the purified product was 75.9% and the radiochemical purity was 99.27%.

[0408] In addition, 24 hours after the production, the samples were analyzed by thin layer chromatography (TLC) in the same manner as above. 211 At-AAMT-OEt. The results are as follows Fig. 9 As shown. Fig. 9 It can be seen that even after 24 hours of purification, no 211 Decomposition of At-AAMT-OEt.

[0409] Example 7

[0410] (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-propyl-α-methyltyrosine hydrochloride ( 24 )(Compound( 24 Synthesis of )Bpin-AMT-OPr)

[0411]

[0412] a)(S)-3-iodo-α-methyltyrosine( 20 )(Compound( 20 Synthesis of

[0413] 7.1 g of iodine (28.1 mmol) was added to a mixture of 5.3 g of potassium iodide (32.1 mmol) and 14 mL of water, and the mixture was stirred for 2 hours. 5.0 g of (S)-α-methyltyrosine ( 19 ) (25.6mmol), 62mL of concentrated ammonia water (28%) and 8mL of water were stirred at -7 to -5°C for 3 hours. 15mL of 15% sodium sulfite aqueous solution was added to the reaction solution, and after warming to room temperature, 6M hydrochloric acid was added under ice-cooling to adjust the pH to 6.5 to 7. After stirring under ice-cooling for 1 hour, the precipitated solid was filtered. After washing with cold water and acetone, it was dried under reduced pressure to obtain 8.4g of compound ( 20 ) (yield 98%).

[0414] 1 H-NMR (400MHz, DMSO-d6+TFA, TMS): 8.25 (3H, br), 7.51 (1H, d, J = 2.0Hz), 7.03 (1H, dd, J = 2.0 ,8.4Hz),6.84(1H,d,J=8.4Hz),3.03(1H,d,J=14.1Hz),2.87(1H,d,J=14.1Hz),1.44(3H,s)

[0415] b) (S)-N-Boc-3-iodo-O-propyl-α-methyltyrosine propyl ester ( 21 )(Compound( 21 Synthesis of

[0416] In 4.2g of compound ( 20)(13.1mmol), 30mL 1M sodium hydroxide aqueous solution (2.3eq.), 25mL water and 50mL 1,4-dioxane mixed solution, add 3.1g di-tert-butyl dicarbonate (14.4mmol, 1.1eq.), stir at 60°C for 22 hours. After cooling the reaction solution to room temperature, concentrate the reaction solution under reduced pressure, add 30mL water and 60mL methyl tert-butyl ether-heptane (1:2), separate the liquid, extract the organic layer with 20mL 5% sodium bicarbonate aqueous solution and 20mL water, combine the aqueous layers, add 0.5M sodium bisulfate aqueous solution, adjust the pH to 1-2, and extract with 60mL methyl tert-butyl ether-ethyl acetate (1:1). After washing the extract with water and saturated brine, distill off the solvent under reduced pressure to obtain 2.1g of light brown solid. The obtained 2.1 g of light brown solid (5.0 mmol) was dissolved in 20 mL of N,N-dimethylformamide, and 1.5 g of potassium carbonate (11 mmol, 2.2 eq.) and 1.1 mL of iodopropane (11 mmol, 2.2 eq.) were added, and stirred at room temperature for 18 hours. 60 mL of ethyl acetate was added, and the solvent was removed by vacuum distillation after washing with 60 mL of water and 30 mL of saturated saline. The residue was purified by silica gel column chromatography (Yamazen 2L size column, ethyl acetate-heptane = 1 / 19 → 1 / 9 → 2 / 8 → 3 / 7 → 5 / 5) to obtain 2.0 g of the compound ( 21 ) (yield 30%).

[0417] 1 H-NMR (400MHz, CDCl3, TMS): 7.50 (1H, d, J = 2.0Hz), 6.98 (1H, dd, J = 2.0, 8.4Hz), 6.68 (1H, d, J = 8.4Hz), 5.18 (1H, brs), 4.18-4.04 (2H, m), 3.33(1H,d,J=13.6Hz),3.11(1H,d,J=13.6Hz),1.88-1.64(4H,m),1.56(3H,s),1.49(9H,s),1.09(3H,t,J=7.2Hz),0.98(3H,t,J=7.2Hz)

[0418] c) (S)-N-Boc-3-iodo-O-propyl-α-methyltyrosine benzyl ester ( 22 )(Compound( 22 Synthesis of

[0419] In the compound ( 21)3.1g (6.1mmol) of methanol 16mL, tetrahydrofuran 16mL, add 2M sodium hydroxide aqueous solution 11.0mL (22.0mmol, 3.6eq.) to the mixture, and stir at 60℃ for 17 hours. After cooling the reaction solution to room temperature, concentrate the reaction solution. Add 30mL of water and 30mL of methyl tert-butyl ether to the residue for separation, add 0.5M sodium hydrogen sulfate aqueous solution to the water layer, adjust the pH to 2-3, and extract with 30mL of methyl tert-butyl ether. After washing the extract with 20ml of water, remove the solvent by vacuum distillation to obtain 2.9g of white amorphous. Dissolve the obtained 2.9g of white amorphous in 16mL of N,N-dimethylformamide, add 1.0g of potassium carbonate (7.4mmol, 1.2eq.) and 1.3mL of benzyl bromide (7.4mmol, 1.2eq.), and stir at 50℃ for 3 hours. The mixture was cooled to room temperature, 30 mL of ethyl acetate was added, and the solvent was removed by distillation under reduced pressure after washing with 30 mL of water and 30 mL of saturated saline. The residue was purified by silica gel column chromatography (Yamazen 2L column, ethyl acetate-heptane = 1 / 9 → 2 / 8 → 3 / 7) to obtain 3.0 g of a white amorphous compound ( 22 ) (yield 89%).

[0420] 1 H-NMR(400MHz,DMSO-d6,TMS):7.44(1H,brs),7.38-7.30(5H,m),7.08(1H,brs ),7.07(1H,dd,J=2.0,8.4Hz),6.89(1H,d,J=8.4Hz),5.11(1H,d,J=12.4Hz),5. 03(1H,d,J=12.4Hz),3.95(2H,t,J=6.4Hz),3.28(1H,d,J=13.2Hz),2.72(1H,d ,J=13.2Hz),1.77-1.68(2H,m),1.42(9H,s),1.18(3H,s),1.02(3H,t,J=7.2Hz)

[0421] d) (S)-N-Boc-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-propyl-α-methyltyrosine benzyl ester ( 23 )(Compound( 23 Synthesis of

[0422] Under nitrogen atmosphere, 3.0 g of compound ( 22)(5.4mmol), 0.34g [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (II) dichloromethane adduct (0.41mmol, 0.076eq.), 2.1g potassium acetate (21.7mmol, 4eq.) and 22mL dimethyl sulfoxide were added with 2.8g of diboric acid pinacol ester (10.8mmol, 2eq.), stirred at room temperature for 1 hour, and then stirred at 55°C for 2 hours. The reaction solution was cooled to room temperature, 50mL of methyl tert-butyl ether and 50mL of water were added, and diatomaceous earth filtration was performed. After separation, the aqueous layer was extracted with 30mL of methyl tert-butyl ether, the organic layers were combined, washed with 30mL of water and 30mL of saturated brine, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (Yamazen 2 L size column, ethyl acetate-heptane = 1 / 19 → 1 / 9 → 2 / 8 → 3 / 7) to obtain 1.5 g of the compound ( 23 ) (yield 50%).

[0423] 1 H-NMR (400MHz, CDCl3, TMS): 7.40-7.32 (6H, m), 7.02 (1H, dd, J = 2.0, 8.4Hz), 6.69 (1H, d, J = 8.4Hz), 5.19 (1H, d, J = 12.4Hz), 5.13 (1H, d, J = 12.4Hz), 3. 89(2H,t,J=6.4Hz),3.28(1H,d,J=13.6Hz),3.16(1H,d,J=13.6Hz),1.85- 1.76(2H,m),1.57(3H,s),1.45(9H,s),1.31(12H,s),1.07(3H,t,J=7.2Hz)

[0424] e) (S)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-O-propyl-α-methyltyrosine hydrochloride ( 24 )(Compound( 24 Synthesis of )Bpin-AMT-OPr)

[0425] 600 mg of compound ( 23) (1.1mmol), 150mg 10% palladium-carbon (55% water) (Pd 1.5 wt%) and 6mL tetrahydrofuran were stirred for 3 hours under hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth and the solvent was distilled off under reduced pressure to obtain 530mg white amorphous. 2mL of 4M hydrochloric acid-ethyl acetate solution was added to the obtained mixture of 530mg amorphous (1.1mmol) and 4.0mL ethyl acetate, and stirred at room temperature for 3 hours. The precipitated solid was filtered and washed with 5.0mL ethyl acetate to obtain 0.18g of the compound as a white powder ( 24 ) (yield 43%).

[0426] 1 H-NMR (400MHz, DMSO-d6, TMS): 8.32 (3H, br), 7.33 (1H, d, J = 2.4Hz), 7.24 (1H, dd, J = 2.4, 8.4Hz), 6.90 (1H, d, J = 8.4Hz), 3-91-3 .85(2H,m),3.08(1H,d,J=14.4Hz),2.97(1H,d,J=14.4Hz),1.74-1.66(2H,m),1.46(3H,s),1.27(12H,s),1.02(3H,t,J=7.2Hz)

[0427] HPLC analysis conditions

[0428] Sample solution: 1 mg of compound ( 24 ) was dissolved in 1 mL of 67% acetonitrile water to prepare a sample solution.

[0429] Detector: UV 275nm

[0430] Chromatographic column: YMC-Pack Pro C18 RS ( 5μm,YMC)

[0431] Column temperature: 30°C

[0432] Mobile phase: H2O (0.1% TFA) / MeCN (0.1% TFA)

[0433] Mode: Gradient

[0434] 0min H2O(0.1%TFA) / MeCN(0.1%TFA)=95 / 5

[0435] 30min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0436] 40min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0437] Flow rate: 1mL / min

[0438] Injection volume: 5μL

[0439] Analysis time: 40 minutes

[0440] Holding time: Compound ( 24 )10.6 minutes

[0441] HPLC chart Fig.10 shown.

[0442] Example 8

[0443] (S)-3-Astatine( 211 At)-O-propyl-α-methyltyrosine ( 211 Synthesis of At-AAMT-OPr

[0444]

[0445] Sodium bicarbonate (7%) was used to dissolve the compound ( 24 ) (Bpin-AMT-OPr), prepare an aqueous solution (1 mg / mL), and place the aqueous solution (0.1 mL) in a 1.5 mL polypropylene (PP) tube. Then, add the 211 At aqueous solution (100 μL, 20 MBq, measured with a Curie meter (IGC-7, Hitachi)) and 0.1 M KI aqueous solution (20 μL) were added, followed by sterile distilled water to make the total volume 200 μL. The mixture was reacted at 50°C for 45 minutes to obtain (S)-3-astatine ( 211 At)-O-propyl-α-methyltyrosine ( 211 At-AAMT-OPr) crude product solution.

[0446] The crude product solution (1 μL) was analyzed by thin layer chromatography (TLC). The sample was applied to a thin layer plate (silica gel 60F 254 ) was used as the developing solvent using a mixture of acetonitrile and water (2 / 1). After the thin layer plate was exposed on an imaging plate (BAS IP MS2025E, GE Healthcare) for about 15 minutes, the plate was analyzed by a bio-image analyzer (Tyhoon TM FLA-7000, Cytiva) was used for analysis. 211 At-AAMT-OPr, the radiochemical yield (RCY) was 78.0%.

[0447] The crude product solution was injected into an HLB column (Oasis, WAT-186005125) equilibrated with sterile distilled water (1 mL), the target product was captured in the cartridge, and the impurities were eluted by passing the sterile distilled water. Next, a 30% ethanol solution (500 μL) was passed through the cartridge, and the target product was eluted into an aqueous sodium ascorbate solution (500 μL, final concentration 1%) to obtain a purified product. The purified product was analyzed by thin layer chromatography (TLC) in the same manner as above. The results are as follows: Fig.11 The radiochemical yield of the purified product was 78.0% and the radiochemical purity was 99.52%.

[0448] In addition, 24 hours after the production, the samples were analyzed by thin layer chromatography (TLC) in the same manner as above. 211 At-AAMT-OPr. The results are as follows Fig.12 As shown. Fig.12 It can be seen that even after 24 hours of purification, no 211 Decomposition of At-AAMT-OPr.

[0449] Example 9

[0450] (S)-α-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-methoxyphenylalanine hydrochloride ( 28 )(Compound( 28 Synthesis of )Bpin-AMPhe-OMe)

[0451]

[0452] a) (S)-N-Boc-α-methyl-4-iodo-3-methoxyphenylalanine benzyl ester ( 26 )(Compound( 26 Synthesis of

[0453] In 1.3g (S)-α-methyl-4-iodo-3-methoxyphenylalanine compound ( 25)(3.9mmol), 8.9mL 1M sodium hydroxide aqueous solution (2.3eq.), 6.5mL water and 13ml 1,4-dioxane mixed solution was added with 930mg di-tert-butyl dicarbonate (4.3mmol, 1.1eq.), and stirred at 60°C for 22 hours. After the reaction solution was cooled to room temperature, it was concentrated under reduced pressure, 30mL water and 60mL methyl tert-butyl ether-heptane (1:2) were added, and the liquid was separated. The organic layer was extracted with 20mL 5% sodium bicarbonate aqueous solution and 20mL water, and the aqueous layer was combined, 0.5M sodium bisulfate aqueous solution was added, and the pH was adjusted to 1-2, and then extracted with 50mL methyl tert-butyl ether-ethyl acetate (1:1). After washing the extract with water and saturated brine, the solvent was distilled off under reduced pressure to obtain 1.3g white amorphous. The obtained 1.3 g of white amorphous was dissolved in 13 mL of N,N-dimethylformamide, and 0.7 g of potassium carbonate (5.1 mmol, 1.2 eq.) and 0.58 mL of benzyl bromide (5.1 mmol, 1.2 eq.) were added, and stirred at 50°C for 3 hours. The mixture was cooled to room temperature, 30 mL of ethyl acetate was added, and the solvent was removed by distillation under reduced pressure after washing with 30 mL of water and 30 mL of saturated saline. The residue was purified by silica gel column chromatography (Yamazen 2 L size column, ethyl acetate-heptane = 1 / 19 → 1 / 9 → 2 / 8) to obtain 1.1 g of a white amorphous compound ( 26 ) (yield 54%).

[0454] 1 H-NMR (400MHz, CDCl3, TMS): 7.57 (1H, d, J = 7.6Hz), 7.41-7.31 (5H, m), 6.53 (1H, d, J = 1.6Hz), 6.36 (1H, dd, J = 1.6, 7.6Hz),5.22-5.13(3H,m),3.77(3H,s),3.36(1H,d,J=13.6Hz),3.23(1H,d,J=13.2Hz),1.59(3H,s),1.44(9H,s)

[0455] b) (S)-N-Boc-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-α-methyl-3-methoxyphenylalanine benzyl ester ( 27 )(Compound( 27 Synthesis of

[0456] Under nitrogen atmosphere, 1.1 g of compound ( 26)(2.1mmol), 0.13g [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (II) dichloromethane adduct (0.16mmol, 0.076eq.), 0.82g potassium acetate (8.4mmol, 4eq.) and 8.4mL dimethyl sulfoxide were added with 1.1g of diboric acid pinacol ester (4.2mmol, 2eq.), stirred at room temperature for 1 hour, and then stirred at 55°C for 1 hour. The reaction solution was cooled to room temperature, 30mL of methyl tert-butyl ether and 30mL of water were added, and diatomaceous earth filtration was performed. After separation, the aqueous layer was extracted with 30mL of methyl tert-butyl ether, the organic layers were combined, washed with 30mL of water and 30mL of saturated brine, and the solvent was removed by distillation under reduced pressure. The residue was purified by silica gel column chromatography (Yamazen 2L size column, ethyl acetate-heptane = 1 / 19→1 / 9→2 / 8) to obtain 0.61g of the compound as a white amorphous ( 27 ) (yield 56%).

[0457] 1 H-NMR (400MHz, CDCl3, TMS): 7.54 (1H, d, J = 7.2Hz), 7.40-7.31 (5H, m), 6.64 (1H, dd, J = 1.2, 7.2Hz), 6.58(1H,d,J=1.2Hz),5.22-5.06(3H,m),3.76(3H,s),3.40-3.21(2H,m),1.63(3H,s),1.43(9H,s)

[0458] c) (S)-α-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-methoxyphenylalanine hydrochloride ( 28 )(Compound( 28 Synthesis of )Bpin-AMPhe-OMe)

[0459] 600 mg of compound ( 27 ) (1.1mmol), 150mg 10% palladium-carbon (55% water) (Pd 1.5 wt%) and 6mL tetrahydrofuran were stirred for 3 hours under a hydrogen atmosphere. The reaction solution was filtered through diatomaceous earth and the solvent was distilled off under reduced pressure to obtain 480mg of white amorphous. 2mL of 4M hydrochloric acid-ethyl acetate solution was added to the obtained mixture of 480mg amorphous (1.1mmol) and 4.0mL ethyl acetate, and stirred at room temperature for 24 hours. The reaction solution was concentrated, 10mL MTBE was added, and then 10mL heptane was added. The precipitated solid was filtered and washed with 5.0mL of a 1 / 1 mixture of MTBE / heptane to obtain 0.14g of the compound as a white powder ( 28 ) (yield 33%).

[0460] 1 H-NMR(400MHz,D2O):7.61(1H,d,J=12.8Hz),6.91(2H,s),3.91(3H,s),3.36(1H,d,J=14.0Hz),3.04(1H,d,J=14.0Hz),1.60(3H,s),1.21(12H,s)

[0461] HPLC analysis conditions

[0462] Sample solution: 1 mg of compound ( 28 ) was dissolved in 1 mL of 67% acetonitrile water to prepare a sample solution.

[0463] Detector: UV 275nm

[0464] Chromatographic column: YMC-Pack Pro C18 RS ( 5μm,YMC)

[0465] Column temperature: 30°C

[0466] Mobile phase: H2O (0.1% TFA) / MeCN (0.1% TFA)

[0467] Mode: Gradient

[0468] 0min H2O(0.1%TFA) / MeCN(0.1%TFA)=95 / 5

[0469] 30min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0470] 40min H2O(0.1%TFA) / MeCN(0.1%TFA)=5 / 95

[0471] Flow rate: 1mL / min

[0472] Injection volume: 5μL

[0473] Analysis time: 40 minutes

[0474] Holding time: Compound ( 28 )7.4 minutes

[0475] HPLC chart Fig.13 shown.

[0476] Example 10

[0477] (S)-α-Methyl-4-astatine( 211 At)-3-methoxyphenylalanine ( 211Synthesis of At-AAMPhe-OMe

[0478]

[0479] By the same method as that described in Example 2, (S)-α-methyl-4-astatine ( 211 At)-3-methoxyphenylalanine ( 211 At-AAMPhe-OMe) crude product solution ( 211 At dosage is 1.525 MBq).

[0480] The crude product solution was analyzed by thin layer chromatography (TLC) in the same manner as described in Example 2. 211 At-AAMPhe-OMe, the radiochemical yield (RCY) was 61.34%.

[0481] The crude product solution was treated with a sterilization treatment and a sodium ascorbate aqueous solution in the same manner as described in Example 2 to obtain a purified product. The purified product was analyzed by thin layer chromatography (TLC) in the same manner as described in Example 2. The results are as follows: Fig.14 The radiochemical yield of the purified product was 65.11% and the radiochemical purity was 97.91%.

[0482] In addition, 24 hours after the production, the samples were analyzed by thin layer chromatography (TLC) in the same manner as above. 211 At-AAMPhe-OMe. The result is Fig.15 As shown. Fig.15 It can be seen that even after 24 hours of purification, no 211 Decomposition of At-AAMPhe-OMe.

[0483] Bpin-AMPhe-OEt and Bpin-AmpPhe-OPr (corresponding boronic acid pinacol esters) can be synthesized by the same or similar method as described in Example 9, and Bpin-AMPhe-OEt and Bpin-AmpPhe-OPr (corresponding boronic acid pinacol esters) can be synthesized from these compounds by the same or similar method as described in Example 2. 211 At-AAMPhe-OEt and 211 At-AAMPhe-Opr (corresponding to 4- 211 At things).

[0484] Experimental Example 1 Cellular Uptake Assay (Using HEK293)

[0485] <Cell culture>

[0486] Cells used: HEK293-Mock, HEK293-hLAT1, HEK293-hLAT2

[0487] (HEK293: Human Embryonic Kidney Cell 293)

[0488] Culture medium used: E-MEM (supplemented with 10% FBS, 1% P / S, 1% NEAA)

[0489] Two days before the experiment, prepare a cell suspension so that the concentration of the cells is 1 x 10 5 Cells / mL. 0.5 mL was inoculated into each well of a 24-well multi-culture plate. Subconfluence was confirmed on the day of the test (as a density, 70-80% of the wells were covered). The cells were washed twice with PBS(-) buffer (166-23555, Fujifilm Wako Pure Chemical Industries, Ltd.), replaced with HBSS(-) buffer (085-09355, Fujifilm Wako Pure Chemical Industries, Ltd.), and left to stand in a culture vessel (MINIcell-35, WakenBtech) until the RI addition test.

[0490] <Uptake test into cultured cells>

[0491] Add 10 μL / well to the prepared cells. 211 At-AAMT-OMe or 211 At-AAMT. BCH (LAT1 inhibitor, 20 mM) or unlabeled substance (α-methyltyrosine, 1 μg / mL) was added in advance (15 minutes before) (10 μg / mL). Culture was performed (CO2 5%, 37 degrees), and after 30 minutes, the liquid was removed by suction using an aspirator. Wash twice with PBS (-). 0.1N NaOH was added to dissolve the cells, and the radioactivity of the dissolved solution was measured using a γ counter. After the radioactivity was measured, 10 μL was measured in each 96-well plate, and the amount of protein was determined by the BCA method, and the number of cells was corrected based on the amount of protein. The absorbance was measured using MultiScan FC (ThermoFisher). Calculation 211 At-AAMT-OMe or 211 The amount of At-AAMT taken into cells (radioactivity counts / amount of protein). Fig.16 shown.

[0492] exist Fig.16 In this study, CTL was used as a control, BCH was used as 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid, and AMT was used as α-methyltyrosine. Fig.16 It can be seen that 211 At-AAMT-OMe 211Similarly, At-AAMT was strongly taken up into HEK293-hLAT1 cells (CTL), and the uptake was significantly inhibited in the presence of BCH and AMT. 211 At-AAMT-Ome was specifically taken up in HEK293-hLAT1 cells.

[0493] Experimental Example 2 Imaging of subcutaneously transplanted mice with human pancreatic cancer cell line PANC1

[0494] Immunodeficient mice (Balb / c-nu / nu, 5-week-old, male) were acclimated for 1 week and the human pancreatic cancer cell line PANC1 was inoculated at 1×10 7 The cells were transplanted subcutaneously. After transplantation, the process was observed until the tumor size reached a size large enough for imaging (1000mm 3 After reaching the desired size, the dilution with water for injection was administered through the tail vein. 211 At-AAMT-OMe or 211 At-AAMT was administered to each mouse to a concentration of 0.5 MBq / 0.2 mL. Isoflurane inhalation anesthesia was performed 2 hours after administration, and planar images were taken using a gamma camera (E-cam, Siemens). Fig.17 shown.

[0495] Depend on Fig.17 It can be seen that 211 At-AAMT-OMe and 211 At-AAMT has been confirmed to accumulate in tumor sites. 211 At-AAMT-OMe was more retained in tumors.

[0496] Experimental Example 3: Therapeutic Experiment of Subcutaneously Transplanted Mice with Human Pancreatic Cancer Cell Line PANC1

[0497] In the same manner as in Experimental Example 2, human pancreatic cancer cell line PANC1 was cultured at 1×10 7 The cells were transplanted subcutaneously into mice. The process was observed until tumor implantation (50-100 mm 3 ), after reaching a predetermined size, the dilution with water for injection was administered from the tail vein 211 At-AAMT-OMe or 211 At-AAMT was administered to each mouse to a concentration of 1MBq / 0.2mL. The mice were euthanized 28 days after administration, and the tumor masses were removed after autopsy to confirm whether there were any abnormalities in the organs. After trimming the adipose tissue, the wet weight of the removed tumor masses was measured and a bar graph was prepared. The results are shown in Figure 2. Fig.18 shown.

[0498] according to Fig.18 , apparently, 211The tumor size ratio of the At-AAMT-OMe administration group was 211 The At-AAMT group was slightly smaller. 211 The antitumor effect of At-AAMT-OMe was slightly higher than 211 At-AAMT.

[0499] in addition, Fig.19 The body weights of mice from immediately after administration to 28 days after administration are shown in FIG. Fig.19 It can be seen that 211 The body weight of mice in the At-AAMT-OMe-administered group tended to decrease slightly, but the weight loss was significantly smaller than that of the control group, and considering the growth of the tumor, it was not to a degree that could be recognized as a side effect.

[0500] Test Example 4 Cellular uptake test (using PANC1)

[0501] <Cell culture>

[0502] Cells used: PANC1

[0503] Culture medium used: D-MEM (supplemented with 10% FBS, 1% P / S)

[0504] As in Experimental Example 1, a cell suspension was prepared 2 days before the test day so that the concentration of the predetermined cells was 1×10 5 Cells / mL. 0.5 mL was inoculated into each well of a 24-well multi-culture plate. Subconfluence was confirmed on the day of the test (as a density, 70-80% of the wells were covered). The cells were washed twice with PBS(-) buffer (166-23555, Fujifilm Wako Pure Chemical Industries, Ltd.), replaced with HBSS(-) buffer (085-09355, Fujifilm Wako Pure Chemical Industries, Ltd.), and left to stand in a culture vessel (MINIcell-35, WakenBtech) until the RI addition test.

[0505] <Uptake test into cultured cells>

[0506] Add 10 μL / well to the prepared cells. 211 At-AAMT-OMe, 211 At-AAMT-OEt, 211 At-AAMT-OPr or 211At-AAMT. BCH (LAT1 inhibitor, 20 mM) or unlabeled substance (α-methyltyrosine, 1 μg / mL) was added in advance (15 minutes before) (10 μg / mL). Culture was performed (CO2 5%, 37 degrees), and after 30 minutes, the liquid was removed by suction using an aspirator. Wash twice with PBS (-). 0.1N NaOH was added to dissolve the cells, and the radioactivity of the dissolved solution was measured using a γ counter. After the radioactivity was measured, 10 μL was measured in each 96-well plate, and the amount of protein was determined by the BCA method, and the number of cells was corrected based on the amount of protein. The absorbance was measured using MultiScan FC (ThermoFisher). Calculation 211 At-AAMT-OMe, 211 At-AAMT-OEt, 211 At-AAMT-OPr or 211 The amount of At-AAMT uptake into cells (radioactivity counts / amount of protein).

[0507] The results are as follows Fig. 20 In addition, Fig. 20 In the above table, “OMe” stands for 211 At-AAMT-OMe, "OEt" means 211 At-AAMT-OEt, "OPr" means 211 At-AAMT-OPr, "OH" means 211 At-AAMT.

[0508] Depend on Fig. 20 It can be seen that 211 At-AAMT-OMe 211 Compared with At-AAMT, it is very superior in terms of intake amount, and the intake amount right after addition is also large, so it is also very superior in terms of intake speed. 211 At-AAMT-OEt and 211 At-AAMT-OPr is similar to 211 At-AAMT is equal to or above. 211 At is a short-lived nuclide, so it can be said that the high uptake into cancer cells in a short period of time is a great advantage.

[0509] Experimental Example 5 Human pancreatic cancer cell line PANC1 was subcutaneously transplanted into mice 211 Determination of the Aggregation Amount of At-Tagged

[0510] Immunodeficient mice (Balb / c-nu / nu, 5-week-old, male) were acclimated for 1 week and the human pancreatic cancer cell line PANC1 was inoculated at 1×10 7 The cells were transplanted subcutaneously. After transplantation, the process was observed until the tumor size was large enough (1000mm 3After reaching the desired size, the dilution with water for injection was administered through the tail vein. 211 At-labeled, to reach 0.1MBq / 0.2mL per mouse. The dose before and after administration was measured, and the difference was used as the administration amount (injected dose: ID). Isoflurane inhalation anesthesia was performed 10 minutes and 1 hour after administration, and the tumors were removed in chronological order and measured using a gamma counter (2480Wizard 2 The dosage was determined by using a semi-microbalance (Shimadzu Corporation, with a minimum display of 10 μg). In order to compare the aggregation amount, the aggregation amount per unit amount (%ID / g) was calculated. The results are shown in Table 1.

[0511] Table 1 shows the PANC1 tumor model 211 Tumor accumulation of At-markers (%ID / g). It should be noted that in Table 1, "OMe" means 211 At-AAMT-OMe, "OEt" means 211 At-AAMT-OEt, "OPr" means 211 At-AAMT-OPr, "OH" means 211 At-AAMT.

[0512] PANC1 tumor model 211 Comparison of tumor accumulation of At-markers (%ID / g)

[0513] <![CDATA[ 211 At-markers]]> OH OMe OE OPr 10 minutes 0.29 2.59 4.21 5.91 1hour 2.49 3.78 3.59 9.45

[0514] It can be confirmed from Table 1 that compared with 211 At-AAMT, 211 At-AAMT-OMe, 211 At-AAMT-OEt and 211 At-AAMT-OPr has high accumulation in tumor sites, among which, 211 The highest aggregation of At-AAMT-OPr was confirmed 1 hour after administration. 211 Compared with At-AAMT, 211 At-AAMT-OMe, 211 At-AAMT-OEt and 211 At-AAMT-OPr, in particular 211 At-AAMT-OPr has excellent retention in tumor sites.

[0515] In addition, 211At-AAMT-OMe was also tested in the same manner as above. The results showed that the tumor accumulation after 10 minutes was 3.77% ID / g, and after 1 hour the tumor accumulation was 6.38% ID / g, which was similar to 211 Compared with At-AAMT, higher accumulation was confirmed at the tumor site.

[0516] Experimental Example 6: Therapeutic Experiment of Subcutaneous Transplantation of Human Pancreatic Cancer Cell Line PANC1 into Mice

[0517] In the same manner as in Experimental Example 5, human pancreatic cancer cell line PANC1 was cultured at 1×10 7 The cells were implanted subcutaneously in mice. The process was observed until tumor implantation (50-100 mm) was confirmed. 3 ), after reaching a predetermined size, the dilution with water for injection was administered from the tail vein 211 At-AAMT-OMe or 211 At-AAMT-OEt was administered to each mouse to achieve 1 MBq / 0.2 mL. The size of the tumor and body weight were measured over time after administration. The experiment was terminated 21 days after administration. The size of the subcutaneous tumor was changed as follows: Fig.21 In addition, Fig. 22 The weight changes of mice from immediately after administration to 21 days after administration are shown in FIG. Fig.21 and Fig. 22 In the above table, “OMe” stands for 211 At-AAMT-OMe, "OEt" means 211 In addition, for the control group, since the tumor size exceeded 10% of the body weight, reaching the humane endpoint, the size change of the subcutaneous tumor and the body weight of the mice after 7 days were Fig.21 and Fig. 22 Not shown.

[0518] according to Fig.21 , apparently, 211 The tumor growth inhibition effect of At-AAMT-OEt administration group was 211 The At-AAMT-OMe administration group showed the same level. In addition, no lesions other than tumors were confirmed in the autopsy results. 3 As a mass, it is equivalent to about 1g. Fig. 22 , it can be inferred that the weight of the control group decreased dramatically, but 211 At-AAMT-OMe administration group, 211 The body weights of the At-AAMT-OEt-administered groups all increased slowly, and no significant toxicity due to the administration was observed.

[0519] Experimental Example 7 Establishment of CDDP-resistant non-small cell lung cancer cell line A549 (A549 CDDP-r)

[0520] Cisplatin (CDDP) was added to the human lung cancer cell line A549 at an initial concentration of 1 μg / mL. After 3 days, dead cells were removed and CDDP was added again. Cells that showed the same proliferation as A549 in the presence of CDDP were separated and maintained while the concentration of CDDP in the culture medium was slowly increased for 3 months. The cell survival rate of CDDP-resistant A549 (A549 CDDP-r) is shown in Figure 2. Fig.23 shown.

[0521] from Fig.23 It was found that CDDP-resistant A549 (A549 CDDP-r) survived at 100 μg / mL. The cells produced by this procedure were used as CDDP-resistant A549 (A549 CDDP-r) in Test Examples 8-10.

[0522] Test Example 8 Cytotoxicity test for CDDP-resistant non-small cell lung cancer cell line A549 (A549 CDDP-r) Test

[0523] Two days before the experiment, A549 and CDDP-resistant A549 (A549 CDDP-r) were cultured at 1×10 5 cells / mL were inoculated into a 96-well plate. 211 At-AAMT-OMe or 211 After adding At-AAMT to each cell, the cells were cultured (CO2 5%, 37 degrees). After 48 hours of culture, Cell counting kit-8 (Dongren Chemical Research Institute) was added. After adding, the cells were cultured at 37 degrees for 1 hour, and the cell survival rate was measured using the absorbance at 450nm as an indicator. The results are shown in Figure 2. Fig.24 shown.

[0524] from Fig.24 It can be seen that 211 At-AAMT also has cytotoxicity against CDDP-resistant A549 (A549 CDDP-r), but 211 The toxicity of At-AAMT-OMe was more significant.

[0525] Experimental Example 9 Treatment of mice subcutaneously transplanted with CDDP-resistant non-small cell lung cancer cell line A549 (A549 CDDP-r) Treatment Trial

[0526] Immunodeficient mice (Balb / c-nu / nu, 5-week-old, male) were acclimated for 1 week and then A549 or CDDP-resistant A549 (A549 CDDP-r) were inoculated at a rate of 1×10 7 The cells were transplanted subcutaneously. The process was observed until tumor implantation (50-100mm) was confirmed. 3 ), after reaching a predetermined size, the dilution with water for injection was administered from the tail vein 211 At-AAMT-OMe or211 At-AAMT was administered to each mouse to a concentration of 0.5 MBq / 0.2 mL. The mice were euthanized 30 days after administration and the tumor masses were removed. After trimming of adipose tissue, the wet weight of the removed tumor masses was measured and a bar graph was prepared. The results are shown in Figure 2. Fig.25 shown.

[0527] Depend on Fig.25 It can be seen that in A549, 211 Antitumor effect of At-AAMT-OMe 211 At-AAMT had the same degree, but in CDDP-resistant A549 (A549 CDDP-r), 211 The anti-tumor effect of At-AAMT-OMe is higher than 211 At-AAMT.

[0528] Experimental Example 10 LAT1 on the cell surface in CDDP-resistant non-small cell lung cancer cell line A549 (A549 CDDP-r) Expression

[0529] A549 and CDDP-resistant A549 (A549CDDP-r) were dispersed using trypsin-EDTA (Fujifilm Wako Pure Chemical Industries, Ltd.). In 0.5% BSA / PBS, an antibody against LAT1 (NOV-NBP2-50465AF647, NOVUSBiologicals) was used for staining, and the expression of LAT1 on the cell surface of A549 and CDDP-resistant A549 (A549CDDP-r) was detected using a flow cytometer (Attune NxT, Thermo Fisher Scientific). The results are shown in Figure 2. Fig.26 shown.

[0530] Depend on Fig.26 It was found that the expression level of LAT1 increased in CDDP-resistant A549 (A549 CDDP-r). Therefore, it was found that the acquisition of anticancer drug resistance is related to the high expression of LAT1.

[0531] Experimental Example 11 Establishment of Gemcitabine-resistant Human Pancreatic Cancer Cell Line PANC1 (PANC1 Gem-r)

[0532] Gemcitabine (Gem) was added to the human pancreatic cancer cell line PANC1 at an initial concentration of 100 ng / mL. Dead cells were removed after 3 days and gemcitabine was added again. Cells that showed the same proliferation as the human pancreatic cancer cell line PANC1 in the presence of gemcitabine were separated and maintained while the concentration of gemcitabine in the culture medium was slowly increased for 3 months. The cell survival rate of the gemcitabine-resistant human pancreatic cancer cell line PANC1 (PANC1Gem-r) is shown in Figure 2. Fig. 27 shown.

[0533] from Fig. 27It was found that the gemcitabine-resistant human pancreatic cancer cell line PANC1 (PANC1 Gem-r) survived at 30 μg / mL. The cells produced by this procedure were used in Test Example 12 as the gemcitabine-resistant human pancreatic cancer cell line PANC1 (PANC1 Gem-r).

[0534] Test Example 12 Treatment of mice subcutaneously transplanted with gemcitabine-resistant human pancreatic cancer cell line PANC1 (PANC1 Gem-r) Treatment Trial

[0535] Immunodeficient mice (Balb / c-nu / nu, 5-week-old, male) were acclimated for 1 week and then inoculated with PANC1 or Gem-resistant PANC1 (PANC1 Gem-r) at a rate of 1×10 7 The cells were implanted subcutaneously. The process was observed until tumor implantation (50-100 mm) was confirmed. 3 ), after reaching a predetermined size, the dilution with water for injection was administered from the tail vein 211 At-AAMT-Ome was administered to each mouse to a concentration of 1 MBq / 0.2 mL. The mice were euthanized 30 days after administration and the tumor masses were removed. After trimming of adipose tissue, the wet weight of the removed tumor masses was measured and a bar graph was prepared. The results are shown in Fig.28 shown.

[0536] Depend on Fig.28 It can be seen that 211 The antitumor effect of At-AAMT-OMe was higher in Gem-resistant PANC1 (PANC1 Gem-r) than that in the parental strain PANC1. 211 At-AAMT-OMe has a higher anti-tumor effect.

[0537] In addition, the tumor size of mice from immediately after administration to 14 days after administration was as follows: Fig.29 The body weight of mice was as shown in Fig.30 It should be noted that for the PANC1-control group, the tumor size exceeded 10% of the body weight (tumor 1000mm 3 Equivalent to 1g. If the mouse is about 20g, the tumor size is greater than 2000mm 3 Therefore, the size change of subcutaneous tumor and body weight of mice after 7 days was Fig.29 and Fig.30 For the same reason, the size change of subcutaneous tumor and body weight of mice in the PANC1-administered group (1 MBq) after 9 days were not shown in the table. Fig.29 and Fig.30 Shown in.

[0538] from Fig.29 and Fig.30 It can be seen that 211 At-AAMT-OMe also showed higher tumor proliferation inhibitory effect in gemcitabine-resistant PANC1 cells.

[0539] [Industrial Applicability]

[0540] According to the present invention, a compound having excellent LAT1 selectivity, higher retention in tumor or cancer sites, and a clearance rate to a degree that does not cause side effects can be provided, thereby being able to exert an improved anti-tumor effect. In addition, a compound that can exert an excellent anti-tumor effect on tumors or cancers that have acquired anticancer drug resistance can also be provided.

[0541] In addition, since the compound is a tyrosine derivative (boronic acid compound (III) and boronic acid compound (II)) into which a boron group (-B(OH)2) or its ester group is introduced, the above-mentioned compound can be produced stably and with good purity by a safe method suitable for industrial production of pharmaceuticals without using harmful substances.

[0542] This application is based on Special Application No. 2022-150608 filed in Japan on September 21, 2022, and all the contents are included in this specification.

Claims

1. A radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof, In formula (I), R 1 Represents C 1-4 alkyl; R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl; R 3 represents a fluorine atom or a chlorine atom; m represents 0, 1 or 2; n represents 0, 1 or 2; X represents the selection 211 At 210 At 131 I. 125 I and 77 Radioactive nuclide of Br.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein m is 1.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein n is 0.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2 C 1-6 alkyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 It is methyl.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2 O is bonded to the 4-position of the benzene ring.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X is bonded to the 3-position of the benzene ring.

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X is 211 At.

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

10. The pharmaceutical composition according to claim 9, further comprising at least one selected from ascorbic acid, an alkali metal ascorbic acid salt and an alkaline earth metal ascorbic acid salt. 11 . A therapeutic agent for tumors or cancers expressing the amino acid transporter LAT1, comprising the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof.

12. The therapeutic agent according to claim 11, wherein: The tumor or cancer expressing the amino acid transporter LAT1 is selected from pancreatic cancer, leukemia, melanoma, colorectal cancer, lung cancer, prostate cancer, gastric cancer, breast cancer, kidney cancer, laryngeal cancer, esophageal cancer, liver cancer, lymphoma, myeloma, head and neck cancer, ovarian cancer, bladder cancer, childhood cancer, childhood leukemia, brain tumor, osteosarcoma, soft tissue sarcoma and bone and soft tissue tumors.

13. The therapeutic agent according to claim 11, wherein The tumor or cancer is a tumor or cancer that has acquired resistance to an anticancer agent.

14. The therapeutic agent according to claim 11, wherein The tumor or cancer is a tumor or cancer that has acquired resistance to platinum preparations.

15. The therapeutic drug according to claim 11, wherein The tumor or cancer is a tumor or cancer that has acquired cisplatin resistance.

16. The therapeutic agent according to claim 11, wherein The tumor or cancer is a tumor or cancer that has acquired resistance to a nucleic acid synthesis inhibitor.

17. The therapeutic agent according to claim 11, wherein The tumor or cancer is a tumor or cancer that has acquired gemcitabine resistance.

18. A compound represented by formula (II) or a salt thereof, In formula (II), R 1 Represents C 1-4 alkyl; R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl; R 3 represents a fluorine atom or a chlorine atom; m represents 0, 1 or 2; n represents 0, 1 or 2; Y represents a boron group (-B(OH)2) or an ester group thereof.

19. The compound or salt thereof according to claim 18, wherein Y is a boronyl group (-B(OH)2) or a 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group.

20. A drug for boron neutron capture therapy, comprising the compound according to claim 18 or 19 or a pharmaceutically acceptable salt thereof.

21. A method for producing a radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof, characterized in that: The following steps are involved: Step 1: In the presence of a reagent selected from the group consisting of alkali metal iodides, alkali metal bromides, N-bromosuccinimide, N-chlorosuccinimide, N-iodosuccinimide and hydrogen peroxide, a compound represented by formula (II) or a salt thereof is reacted in water with a 211 At 210 At 131 I. 125 I, and 77 A step of obtaining a radiolabeled compound represented by formula (I) or a pharmaceutically acceptable salt thereof by reacting a radionuclide of Br with Br, In the formula, R 1 Represents C 1-4 alkyl; R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl; R 3 represents a fluorine atom or a chlorine atom; m represents 0, 1 or 2; n represents 0, 1 or 2; X represents the selection 211 At 210 At 131 I. 125 I and 77 Radionuclides of Br; Y represents a boron group (-B(OH)2) or an ester group thereof.

22. A compound represented by formula (III): In formula (III), R 1 Represents C 1-4 alkyl; R 2 Represents C 1-6 Alkyl, C 1-6 Haloalkyl or C 7-14 Aralkyl; R 3 represents a fluorine atom or a chlorine atom; m represents 0, 1 or 2; n represents 0, 1 or 2; Y represents a boron group (-B(OH)2) or an ester group thereof; P 1 represents a carboxyl protecting group; P 2 represents an amino protecting group.

23. The compound or salt thereof according to claim 22, wherein P 1 is benzyl, and P 2 It is tert-butyloxycarbonyl.

24. A compound represented by formula (IV'), In formula (IV'), R 2a C 1-3 Alkyl, P 1a Represents benzyl or C 1-3 alkyl.

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