JP5476541B2 - Telomerase inhibitor - Google Patents

Description

  The present invention relates to a novel compound having a macrocyclic heptaoxazole structure having an amino group in the side chain. The present invention also relates to a telomerase inhibitor and an antitumor agent comprising the compound and a method for producing the compound. Furthermore, the present invention provides a tool for easily detecting the presence of a G-quadruplex structure, which is a higher order structure of DNA, by fluorescence.

Telomeres are structures that exist at the ends of eukaryotic chromosomes. In the case of humans, there are approximately 10,000 base pairs in a 6-base repetitive sequence (TTAGGG) n, and the 3 ′ end protrudes (overhangs) by approximately 100 bases to form a single strand.
The telomeric overhanging single strand forms a unique helical higher-order structure called G-quadruplex under conditions where monovalent cations such as potassium and sodium ions are present in high concentrations. Yes. It has recently been clarified that this G-quadruplex structure exists not only in telomeres but also in gene promoter regions such as c-kit, c-myc and bcl-2, and has a unique structure. For example, the telomeric G-quadruplex has an antiparallel structure, whereas the c-myc and c-kit promoter regions have a forward parallel structure, and the bcl-2 promoter region has a forward parallel structure. It has a structure with mixed antiparallel styles.

  These G-quadruplex structures are involved in important biological mechanisms. For example, the formation of G-quadruplex significantly shortens telomeres and subsequently causes apoptosis of cancer cells by dissociating TRF2 and / or Pot1 bound to the telomeres. In vitro experiments have shown that the transcriptional activity of c-kit and c-myc is suppressed by the formation of a G-quadruplex structure in the promoter region. Therefore, a substance that binds strongly and specifically to G-quadruplex is not only a biological tool, but can also be a candidate for an anti-cancer agent, so the aim was to develop a strong G-quadruplex binder There is a lot of research.

Telomestatin has a macrocyclic structure with five oxazoles, two methyl oxazoles and one thiazole, and was isolated from Streptomyces anulatus 3533-SV4 by screening with a TRAP (telomeric repeat amplification protocol) assay. Natural organic compounds (Patent Document 1 and Non-Patent Document 1). Telomestatin has a very high telomerase inhibitory activity with an IC ₅₀ of 5 nM, and its structural formula is as follows.

It has been reported that a macrocyclic structure consisting of oxazole and thiazole of telomestatin interacts with telomeres and strongly stabilizes the G-quadruplex structure (Non-patent Document 2). Therefore, telomestatin is widely used in various functional studies as a standard G-quadruplex binder. Studies using telomestatin as an antitumor agent have also been conducted (Non-patent Document 3). However, since telomestatin is a natural organic compound and cannot be obtained in large quantities, research on synthetic methods has been conducted (Patent Document 2 and Non-Patent Document 4).
There have also been many studies on the synthesis and activity evaluation of telomestatin derivatives and analogs (Patent Documents 3 and 4 and Non-Patent Documents 5 to 12).

International Publication WO 00/24747 Pamphlet International Publication WO 02/48153 Pamphlet JP 2006-316008 A International Publication WO 2007/127173 Pamphlet

Shin-ya, K. et al. And 7 others, published in 2001, Journal of the American Chemical Society, vol. 123, pp. 1262-1263 Kim, M. Y. et al. And 4 others, published in 2002, Journal of the American Chemical Society, vol.124, pp.2098-2099 Tauchi, T. et al. And 6 others, published in 2006, Oncogene, vol.25, pp.5719-5725 Doi, K. et al., 3 others, published in 2006, Organic Letters, vol.8, Issue 18, pp.4165-4167 Dairin Ishizuka et al. And 5 others, published on March 12, 2008, “Synthesis and Activity Evaluation of Macrocyclic Heptoxazole (7OTD) Derivatives with Telomestatin as Lead”, Proceedings of the Chemical Society of Japan, Vol.88, No. 2, pp. 1049 Masayuki Tera et al., March 12, 2008, “Synthesis of macrocyclic hexaoxazole (6OTD) derivatives with telomestatin as lead and evaluation of G-quadruplex stabilization”, Proceedings of the Chemical Society of Japan, Vol. 88, No.2, pp.1048 Dairin Ishizuka et al., 4 others, published on December 1, 2007, “Synthetic research and activity evaluation of G-q ligand with telomestatin as lead”, Kanto Branch Symposium of Synthetic Organic Chemistry, Vol.54, pp.56 Dairin Ishizuka et al., 3 others, published on March 12, 2007, Proceedings of the Chemical Society of Japan, Vol.87, No.2, pp.1190 Suzanne G. Rzuczek et al., 3 others, published in 2008, Bioorganic & Medicinal Chemistry Letters, vol.18, pp.913-917 Gurpreet Singh Minhas et al., 4 others, 2006, Bioorganic & Medicinal Chemistry Letters, vol.16, pp.3891-3895 Masayuki Tera and 6 others, published in 2006, Heterocycles, vol.69, pp.505-514 Nobuaki Endoh et al. And 4 others, published in 2003, Heterocycles, vol.60, pp.1567-1572

  In view of the above-described conventional situation, an object of the present invention is to provide a telomestatin analog having high telomerase inhibitory activity and a method for synthesizing the telomestatin analog.

  In order to solve the above problems, the present inventors have intensively studied, and as a result, improved planarity as a macrocyclic structure having seven oxazoles instead of six, and further providing a side chain having an amino group, thereby providing telomerase. The inventors found that a telomestatin analog having a high inhibitory activity can be obtained, and completed the present invention.

  That is, the novel compound according to the present invention includes the following compounds (I) to (VIII).

(I) A compound represented by the following general formula (1).

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom. )

(II) X is —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO— or —NHCONHC (NH) —. The compound according to (I) above, characterized by:

(III) The compound according to (II) above, wherein X is —CH ₂ —, —NHC (NH) — or —CH (NH ₂ ) —.

(IV) In the compound described in (III) above, X is —CH ₂ —, R ₁ is a linear alkylene group having 1 to 7 carbon atoms, and is represented by the following general formula (2): Compound.

(In formula, n represents the integer of 2-8.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom. )

(V) The compound according to (IV) above, wherein n is 4.

(VI) R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently a hydrogen atom, an alkyl group having 1 to 7 carbon atoms which may have an amino group or a guanidino group. The compound according to any one of (I) to (V) above, wherein

(VII) The compound as described in (V) above, wherein any of R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ is a hydrogen atom.

(VIII) In the compound described in the above (I), X is —CH ₂ —, R ₁ is an alkylene group having 3 carbon atoms, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R _7. A compound represented by the following structural formula (3), wherein ₇ is a hydrogen atom.

  Moreover, the novel compound according to the present invention described above can also be provided as a salt. Furthermore, the novel compound or salt thereof according to the present invention described above has applications as, for example, a telomerase inhibitor. Furthermore, the above-described novel compounds or salts thereof according to the present invention have uses as therapeutic agents for diseases based on telomerase inhibitory action. Examples of diseases based on telomerase inhibitory activity include cancer. That is, the novel compound or a salt thereof according to the present invention has a use as an antitumor agent.

  On the other hand, this invention provides the manufacturing method (IX)-(X) of the novel compound based on this invention mentioned above.

(IX) A method for producing the compound described in (I) above, which comprises the following steps (F) to (H):
(F) By reacting a compound represented by the following general formula (8) with methanesulfonyl chloride (MsCl) and triethylamine (Et ₃ N) in dichloromethane, and further adding diazabicycloundecene (DBU), A step of obtaining a compound represented by the following general formula (9) by intramolecular dehydration;

(In the formula, X represents —CH ₂ —, —NHC (NH) — or —CH (NH ₂ ) —).
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )
(G) In an aprotic polar solvent, N-bromosuccinimide (NBS) and cesium carbonate (Cs ₂ CO ₃ ) are allowed to act on the compound represented by the following general formula (9) to obtain the following general formula ( A step of obtaining a compound shown in 10);

(In the formula, X represents —CH ₂ —, —NHC (NH) — or —CH (NH ₂ ) —).
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )
(H) Protecting groups Y and R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ included in the compound represented by the following general formula (10) are removed. The process of making it leave | separate and obtaining the compound shown by following General formula (1).

(In the formula, X represents —CH ₂ —, —NHC (NH) — or —CH (NH ₂ ) —).
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms or a halogen Represents an atom.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )

(X) The production according to (IX) above, wherein the protecting group Y is a tert-butoxycarbonyl group (Boc group) and the protecting group Y (Boc group) is eliminated in an acidic solvent. Method.

  Furthermore, this invention can provide the probe (XI) which visualizes a G-quadruplex structure, and its manufacturing method (XII) by utilizing the novel compound which concerns on this invention.

(XI) A probe for visualizing a G-quadruplex structure obtained by reacting the compound according to any one of (I) to (VIII) above and a fluorescent compound capable of reacting with an amino group in the compound.

  As a specific example of the probe according to the present invention, the compound represented by the general formula (16) is used as the fluorescent compound, and the compound represented by the general formula (17) can be exemplified.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Q represents a fluorescent dye.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms or a halogen Represents an atom.
R ₁₀ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms. )

(XII) A method for producing a probe for visualizing a G-quadruplex structure in which the compound according to any one of (I) to (VIII) above is reacted with a fluorescent compound capable of reacting with an amino group in the compound.

  As an example, the production method according to the present invention specifically includes a reaction of the compound (1) described in the above (I) with the fluorescent compound represented by the following general formula (16) as shown in the following formula. be able to.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Q represents a fluorescent dye.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms or a halogen Represents an atom.
R ₁₀ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms. )

  The compound according to the present invention has a macrocyclic structure having seven oxazoles having high planarity, and has a side chain having an amino group, and thus has a high telomerase inhibitory activity.

  In addition, the method for producing a compound according to the present invention enables a very difficult reaction of ring-closing a part of a compound having six oxazoles to form a macrocyclic structure having seven oxazoles, and has a telomerase activity. A high compound according to the present invention can be produced.

  Furthermore, since the probe of the present invention has an activity to bind to a G-quadruplex structure and can be detected by fluorescence, the presence of the G-quadruplex structure in a living cell, tissue or nucleic acid can be easily detected. It can be used as a tool.

It is a figure which shows the result of having evaluated the functionality of the probe which visualizes the G-quadruplex structure of this invention. In the figure, “+” indicates that 1 equivalent of compound is added to DNA, and “++” indicates that 10 equivalent of compound is added to DNA. In addition, “L1BOD-7OTD” is a photograph of the emission of the gel when excited at the fluorescence wavelength of BODIPY, and “Stains-all” is an image of the emission of the gel when excited at the fluorescence wavelength of BODIPY. It is a photograph and “Merged” is a photograph in which these two are superimposed.

  Hereinafter, modes for carrying out the present invention will be described. The present invention is not limited to the following embodiment. This specification includes the contents described in the specification and / or drawings of Japanese Patent Application No. 2008-166494, which is the basis of the priority of the present application. All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

1. Compounds of the invention
1-1. Structure of the Compound of the Present Invention The compound of the present invention is a compound represented by the following general formula (1). In the general formula and structural formula of the present invention, when optical isomers are present, all of the optical isomers are included.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom. )

1-2. Characteristic Structure of the Compound of the Present Invention and Effects thereof The compound of the present invention has a macrocyclic structure composed of seven oxazoles, and therefore has higher planarity than a compound having six oxazoles. The reason for the high telomerase inhibitory activity of the compound of the present invention is not clear, but it is considered that this high planarity strongly interacts with G-quadruplex.
The compounds of the present _invention has a side chain consisting of -R 1 -X-NH _2. The reason for the high telomerase inhibitory activity of the compound of the present invention is not clear, but it is considered that the amino group of this side chain interacts with a phosphate of a nucleotide forming a G-quadruplex.

The compound of the present invention having a side chain with an amino group but having six oxazoles constituting a macrocyclic structure (Comparative Example 1), and having seven side chains with an amino group and seven oxazoles When the telomerase inhibitory activity was compared, the compound of the present invention exhibited a higher telomerase inhibitory activity (see Comparative Example 1 and Table 1).
Further, telomerase inhibitory activity of a compound having 7 oxazoles but having no amino acid side chain (Comparative Example 2) and a compound of the present invention having an amino group side chain and 7 oxazoles is shown. When compared, the compound of the present invention exhibited higher telomerase inhibitory activity (see Comparative Example 2 and Table 1).

  In addition, the compound of the present invention specifically acted on tumor cells expressing telomerase and exhibited the activity of suppressing tumor growth (see Example 3).

1-3. Structure of the side chain of the compound of the present invention The compound represented by the general formula (1) of the present invention has a side chain composed of -R ₁ -X-NH ₂ , wherein X is -CH _2- , _{-NHC (NH) -, - CH} (NH 2) -, - C (NH) -, - NHCO -, - CO -, - NHCONHC (NH) -, cyclohexadienyl Eni alkylene group, and, 3, 4, 5 , 6-tetrahydropyrimidin-2,6-yl group.
The above divalent group is represented by the structural formula as follows.

When X is —CH ₂ —, —NHCO—, —CO—, a cyclohexadienylene group, or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group, the side chain represents 1 amino group. Side chains. Further, when X is —NHC (NH) — or —NHCONHC (NH) —, the side chain is a side chain having a guanidino group. The guanidino group is a substituent represented by —NHC (NH) NH ₃ . When X is —CH (NH ₂ ) —, the side chain is a side chain having two amino groups.
X is preferably —CH ₂ — or —NHC (NH) —, and particularly preferably —CH ₂ —.

R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
Here, the term “single bond” means that there is no group. For example, when R ₁ is a single bond and X is —CH ₂ —, the side chain is an aminomethyl group.
The alkylene group having 1 to 12 carbon atoms includes not only a linear group but also an alkylene group having a branched chain. Examples of the alkylene group having 1 to 12 carbon atoms include, but are not limited to, methylene, ethylene, trimethylene, 1-methyltrimethylene, 2-methyltrimethylene, 2,2-dimethyltrimethylene, tetra Examples include methylene, 2-methyltetramethylene, 2,3-dimethyltetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene, nonamethylene, decamethylene, undecamethylene, and dodecamethylene.

  The alkenylene group having 1 to 12 carbon atoms includes not only a linear group but also an alkenylene group having a branched chain. Two or more double bonds may be present. Examples of the alkenylene group having 1 to 12 carbon atoms include, but are not limited to, vinylene, propenylene, 1-methyl-1-propenylene, 2-methyl-1-propenylene, and 3-methyl-1-propenylene. 2,3-dimethyl-1-propenylene, 1-butenylene, 2-butenylene, 1,3-butadienylene, 2-pentenylene, 2,3-pentadienylene, 2-hexenylene, 1,3,5-hexatriylene, 3, -Heptenylene, 4-octenylene, 4-nonenylene, 5-decenylene, 6-undecenylene, 6-dodecenylene group, etc. are mentioned.

  Similarly, the alkynylene group having 1 to 12 carbon atoms includes not only a linear group but also an alkynylene group having a branched chain. There may be two or more triple bonds. Examples of the alkynylene group having 1 to 12 carbon atoms include, but are not limited to, ethynylene, propynylene, 3-methyl-1-propynylene, butynylene, 1,3-butadienylene, 2-pentynylene, and 2-pentynylene. 2,4-pentadinylene, 2-hexynylene, 1,3,5-hexatriinylene, 3-heptynylene, 4-octynylene, 4-noninylene, 5-decynylene, 6-undecynylene, 6-dodecynylene, and the like.

R ₁ is preferably an alkylene group having 1 to 12 carbon atoms, more preferably a linear alkylene group having 1 to 7 carbon atoms.
Then, X is -CH 2 _- and, when R ₁ is a straight-chain alkylene group having 1 to 7 carbon atoms, the compounds of the present invention is represented by the following general formula (2).

(In formula, n represents the integer of 2-8.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently a hydrogen atom, an amino group or a guanidino group, an alkyl group having 1 to 6 carbon atoms, an amino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms or a halogen Represents an atom. )

In the said General formula (2), n is an integer of 2-8 and a side chain turns into a C2-C8 aminoalkyl group.
More preferably, n is an integer of 3 to 5, and the side chain is an aminoalkyl group having 3 to 5 carbon atoms. Most preferably, n is 4 and the side chain is an aminobutyl group.

1-4. Peripheral part in the macrocyclic structure of the compound of the present invention In the peripheral part of the macrocyclic structure of the compound of the present invention, that is, the compound represented by the general formula (1) or (2) of the present invention, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently a hydrogen atom, an amino group or a guanidino group, an alkyl group having 1 to 12 carbon atoms, an amino group, a guanidino group or an amino group. Alternatively, it represents an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group, a phenyl group which may have an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen atom.

Here, the amino group is —NH ₂ and the guanidino group is —NHC (NH) —NH ₂ .
In addition, the alkyl group having 1 to 12 carbon atoms which may have an amino group or a guanidino group includes not only a linear group but also an alkylene group having a branched chain. Or you may have two or more guanidino groups in total.
Examples of the alkyl group having 1 to 12 carbon atoms which may have an amino group or a guanidino group include, but are not limited to, for example, methyl group, ethyl group, propyl group, isopropyl group, 3-amino group. Propyl group, 2-aminopropyl group, 2,3-diaminopropyl group, 2-amino-3-guanidinopropyl group, butyl group, 4-aminobutyl group, 3,4-diaminobutyl group, 2-guanidino-4- Aminobutyl group, 4-amino-3- (2-aminoethyl) butyl group, tert-butyl group, pentyl group, 5-aminopentyl group, 5-guanidinopentyl group, hexyl group, 6-aminohexyl group, octyl group , Nonyl group, dodecyl group, undecyl group, dodecyl group and the like.

The aralkyl group having 7 to 12 carbon atoms which may have an amino group or a guanidino group is an alkyl group having 7 to 12 carbon atoms substituted with a monocyclic or condensed cyclic aryl group. You may have not only one group or guanidino group but two or more. Of course, an aralkyl group having no amino group or guanidino group may be used.
Examples of the aralkyl group having 7 to 12 carbon atoms that may have an amino group or a guanidino group include, but are not limited to, benzyl group, 4-aminobenzyl group, 3-guanidino-4-aminobenzyl. Group, phenethyl group, 4-aminophenethyl group, phenylpropyl group, 3,4-diaminophenethyl group, phenylbutyl group, phenylpentyl group, naphthylmethyl group, naphthylethyl group and the like.

  The phenyl group which may have an amino group or a guanidino group includes, but is not limited to, a phenyl group having a total of two or more amino groups and / or guanidino groups. For example, a phenyl group, 4-aminophenyl group, 4-guanidinophenyl group, 3-guanidino-4-aminophenyl group and the like can be mentioned.

Examples of the heterocyclic group having 2 to 6 carbon atoms include, but are not limited to, thienyl group, furyl group, pyranyl group, pyrrolyl group, imidazolyl group, isothiazolyl group, isoxazolyl group, pyridyl group, pyrazinyl group, pyrimidinyl group. Group, pyridazinyl group and the like.
Moreover, as a halogen atom, a fluorine atom, a chlorine atom, and a bromine atom are mentioned, for example.

As one preferred embodiment of the present invention, in the compound represented by the general formula (1) or (2) of the present invention, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently , A hydrogen atom, an amino group or a guanidino group, which may have a C 1-7 alkyl group.
Here, the alkyl group having 1 to 7 carbon atoms which may have an amino group or a guanidino group includes not only a linear group but also an alkylene group having a branched chain. Or you may have two or more guanidino groups in total.
Examples of the alkyl group having 1 to 7 carbon atoms which may have an amino group or a guanidino group include, but are not limited to, for example, methyl group, ethyl group, propyl group, isopropyl group, 3-amino Propyl group, 2-aminopropyl group, 2,3-diaminopropyl group, 2-amino-3-guanidinopropyl group, butyl group, 4-aminobutyl group, 3,4-diaminobutyl group, 2-guanidino-4- Aminobutyl group, 4-amino-3- (2-aminoethyl) butyl group, tert-butyl group, pentyl group, 5-aminopentyl group, hexyl group, 6-aminohexyl group, 6-guanidinohexyl group, heptyl group , 7-aminoheptyl group and the like.

As one preferred embodiment of the present invention, in the compound represented by the general formula (1) or (2) of the present invention, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently , A straight-chain alkyl group having 1 to 7 carbon atoms having a hydrogen atom or an amino group at its end.
Examples of the linear alkyl group having 1 to 7 carbon atoms terminated with an amino group include an aminomethyl group, a 2-aminoethyl group, a 3-aminopropyl group, a 4-aminobutyl group, a 5-aminopentyl group, 6 -An aminohexyl group and a 7-aminoheptyl group are mentioned.

As one preferred embodiment of the present invention, in the compound represented by the general formula (1) or (2) of the present invention, R ₂ , R ₄ , R ₅ and R ₇ are each a hydrogen atom, and R ₃ and R ₆ At least one of them may be an alkyl group having 1 to 7 carbon atoms which may have an amino group or a guanidino group.
As one preferred embodiment of the present invention, in the compound represented by the general formula (2), R ₂ , R ₄ , R ₅ and R ₇ are hydrogen atoms, and both R ₃ and R ₆ are amino-terminal. It can be set as a C1-C7 linear alkyl group which has group.
In this case, the compound of the present invention is represented by the following general formula (12).

(In the formula, n represents an integer of 2 to 8, m represents an integer of 1 to 7, and l represents an integer of 1 to 7.)

As one preferred embodiment of the present invention, in the compound represented by the general formula (1) or (2) of the present invention, R ₂ and R ₃ are methyl groups, and R ₄ , R ₅ , R ₆ and R ₇ are By using a hydrogen atom, a compound having a macrocyclic structure composed of two methyloxazoles and five oxazoles can be obtained in the same manner as natural telomestatin.

As one preferred embodiment of the present invention, in the compound represented by the general formula (1) or (2) of the present invention, R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are hydrogen atoms. be able to.

1-5. Compound represented by the structural formula (3) As one preferred embodiment of the present invention, in the compound represented by the general formula (2), R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each a hydrogen atom. And n can be 4.
In this case, the compound of the present invention is represented by the following structural formula (3).

The compound represented by the structural formula (3) was measured for telomerase inhibitory activity by the method described in Example 2 below. As a result, the compound has a very high telomerase inhibitory activity with an IC ₅₀ of 0.8 nM. It exhibits an activity several times stronger than the telomerase inhibitory activity (IC ₅₀ = about 5 nM) of telomestatin, which is an organic compound.

2. Salts containing compounds of the present invention, telomerase inhibitors and antitumor agents The compounds of the present invention can be salts. For example, a salt can be formed by adding an acid to the compound of the present invention. Examples of acids include, but are not limited to, hydrobromic acid, hydroiodic acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, apple Examples include acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, aspartic acid, and glutamic acid.

Since the compound of the present invention has strong telomerase inhibitory activity, it can be used as a telomerase inhibitor. The telomerase inhibitor of the present invention can be produced, for example, by diluting the compound of the present invention in a solvent such as dimethyl sulfoxide, methanol, water, phosphate buffer, or Tris-HCl buffer that can stably store the compound. it can.
The telomerase inhibitor of the present invention can be widely used in experimental systems that suppress the enzyme activity of telomerase, such as studies on cancer mechanisms and animal and plant development mechanisms.

  On the other hand, since the compound of the present invention has strong telomerase inhibitory activity, it can be used as a therapeutic and / or prophylactic agent for diseases caused by increased telomerase activity.

Examples of the disease caused by increased telomerase activity include cancer.
Note that telomestatin, which is a known telomerase inhibitor, is known to have antitumor activity (Non-patent Document 3: Tauchi, T. et al., Et al., 2006, Oncogene, vol. 25, pp. 5719-5725). Therefore, the antitumor agent containing the compound of the present invention as an active ingredient can be used as an active ingredient of an antitumor agent having telomerase inhibitory activity in the same manner as telomestatin.
As shown in the examples described later, the compound of the present invention has a significantly superior telomerase inhibitory action as compared with telomestatin, and therefore the antitumor agent according to the present invention is more likely to exhibit superior antitumor activity. It can be expected to have an excellent effect on cancer treatment and cancer prevention. In addition, as shown in the examples described later, the antitumor agent according to the present invention specifically acts on tumor cells that highly express telomerase, and thus does not act on normal cells and has few side effects. It is possible to develop agents.

Here, the tumor means a tissue mass in which cells proliferate excessively against the control mechanism of the living body. In the present invention, the cancer is not particularly limited as long as it is a disease accompanied by a tumor. For example, stomach cancer, leukemia, colon cancer, lung cancer, liver cancer, esophageal cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, ovarian cancer , Bladder cancer, prostate cancer, testicular tumor, osteosarcoma, malignant lymphoma, cervical cancer, skin cancer, brain tumor and the like.
The antitumor agent of the present invention can be made into various pharmaceutical dosage forms according to the purpose. For example, by making into tablets, capsules, granules, solutions, emulsions, etc., oral preparations, suppositories, ointments Preparations, patches, eye drops, injections, aerosols.

3. Process for producing the compound of the present invention
3-1. Synthetic Scheme of Compound of the Present Invention The compound of the present invention can be synthesized , for example, according to the following synthetic scheme of the present invention consisting of reactions (C-1) to (H).

(In the reaction formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
Z represents an amino-protecting group different from Y.
W represents a protecting group for a hydroxy group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom.
R ₈ and R ₉ each independently represents a methyl group or an ethyl group.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. An alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group or a guanidino group having a protecting group, an aralkyl group having 7 to 12 carbon atoms, It represents a phenyl group optionally having an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom. )
X in the above scheme represents the same as described above in this specification.

Y in the above scheme represents an amino-protecting group. Examples of Y include, but are not limited to, tert-butoxycarbonyl group, benzyloxycarbonyl group, 9-fluorenylmethyloxycarbonyl group, 2,2,2-trichloroethoxycarbonyl group, allyloxy A carbonyl group etc. are mentioned.
Among these, it is particularly preferable to use a tert-butoxycarbonyl group as Y.

Z in the above scheme represents an amino-protecting group. However, Z is a protecting group different from Y. This is because only one of the protecting groups is eliminated in the reaction (C-2).
Examples of Z include, but are not limited to, benzyloxycarbonyl group, tert-butoxycarbonyl group, 9-fluorenylmethyloxycarbonyl group, 2,2,2-trichloroethoxycarbonyl group, allyloxy A carbonyl group etc. are mentioned.
Among these, it is particularly preferable to use a benzyloxycarbonyl group as Z.

W in the above scheme represents a hydroxyl-protecting group. Examples of W include, but are not limited to, tert-butyldimethylsilyl group, p-methoxybenzyl group, tert-butyl group, acetyl group, benzoyl group, trimethylsilyl group, triethylsilyl group, triisopropylsilyl. Group, tert-butyldiphenylsilyl group and the like.
Among these, it is particularly preferable to use a tert-butyldimethylsilyl group as W.

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the above scheme represent the same as previously described in this specification.
R ₈ and R ₉ in the above scheme each independently represent a methyl group or an ethyl group.
In the above scheme, R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′, and R ₇ ′ are amino groups of R ₂ , R ₃ , R ₄ , R ₅ , R _6, and R ₇ , respectively. The group is protected with a protecting group Y or other protecting group. That is, when Rx (X = 1 to 7) has an amino group or a guanidino group, a group in which the amino group or guanidino group has an amino group protected with a protecting group Y or another protecting group is Rx ′ (X = 1 to 7).

3-2. Synthesis of linear hexaoxazole In the synthesis scheme of the present invention, linear hexaoxazole is synthesized by reactions (C-1) to (C-3).
First, the reaction (C-1) in the synthesis scheme of the present invention performs the following reaction.

(In the reaction formula, Z represents an amino-protecting group.
W represents a protecting group for a hydroxy group.
R ₈ represents a methyl group or an ethyl group.
R ₅ ′, R ₆ ′ and R ₇ ′ each independently represents a carbon atom which may have a hydrogen atom, Y (amino-protecting group) or an amino group or guanidino group protected by another protecting group. An alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; A phenyl group optionally having an amino group or guanidino group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom; )

Said reaction (C-1) is reaction which deprotects Z, and reaction conditions differ with the kind of Z.
For example, when Z is a benzyloxycarbonyl group, the benzyloxycarbonyl group can be deprotected in the presence of a palladium catalyst in a protic polar organic solvent.
Here, the protic polar solvent is not limited to these. For example, a mixed solvent of methanol / THF (tetrahydrofuran), a mixed solvent of ethanol / THF, and the like can be used.
Examples of the palladium catalyst include, but are not limited to, palladium / carbon, palladium hydroxide / carbon, and the like.

Further, when Z is a 9-fluorenylmethyloxycarbonyl group, it can be deprotected with a secondary amine.
When Z is a 2,2,2-trichloroethoxycarbonyl group, it can be deprotected by acting zinc powder-acetic acid or the like.
When Z is an allyloxycarbonyl group, it can be deprotected by adding an amine in the presence of a palladium catalyst.

  Next, reaction (C-2) in the synthetic scheme of this invention performs the following reaction.

(In the reaction formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
Z represents an amino-protecting group different from Y.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₉ represents a methyl group or an ethyl group.
R ₅ ′, R ₆ ′ and R ₇ ′ are each independently a hydrogen atom, an amino group having a protecting group or an alkyl group having 1 to 12 carbon atoms which may have a guanidino group, or an amino having a protecting group. A guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group, an amino group having a protecting group or a guanidino group A phenyl group, a C2-C6 heterocyclic group, or a halogen atom is represented. )

Said reaction (C-2) is reaction which makes alkoxycarbonyl group COOR ₉ carbonyl group COOH, and is performed by making it react with a strongly basic inorganic salt in a hydrous polar solvent.
Here, the hydrous polar solvent is not limited to these, but for example, a mixed solvent of THF and water, a mixed solvent of methanol and water, or the like can be used.
Moreover, as a strongly basic inorganic salt, although not necessarily limited to these, lithium hydroxide, sodium hydroxide, potassium hydroxide etc. can be used, for example.

  The reaction (C-3), which is the final step for synthesizing the linear hexaoxazole, involves the following reaction.

(In the reaction formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
Z represents an amino-protecting group different from Y.
W represents a protecting group for a hydroxy group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₈ represents a methyl group or an ethyl group.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; It represents a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )

The reaction (C-3) is a reaction for condensing the amino group of the compound represented by the general formula (4 ′) and the carboxyl group of the compound represented by the general formula (5 ′).
In this reaction, a condensing agent 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methylmorpholinium (DMT-MM) or a salt thereof is added in a solvent. Can be done. Here, the solvent is not particularly limited, and water, methanol, ethanol, isopropyl alcohol, or a mixed solvent thereof can be used.
Alternatively, by adding 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), N, N′-dicyclohexylcarbodiimide (DCC) or diisopropylcarbodiimide (DIPC) in an aprotic polar solvent. Can be condensed.
Here, examples of the aprotic polar solvent include, but are not limited to, N, N-dimethylformamide (DMF), tetrahydrofuran (THF), dioxane, and the like.

3-3. Macrocyclization of linear hexaoxazole Next, macrocyclization of linear hexaoxazole is performed by reactions (D-1) to (D-3).
Here, the order in which the reactions (D-1), (D-2), and (D-3) are performed, (D-1) may be performed first, or (D-2) may be performed first. Good. Moreover, you may perform reaction (D-1) and (D-2) simultaneously.
That is, (D-1) is a reaction for deprotecting the protecting group Z of the —NHZ group at the end of the compound, and (D-2) is the alkoxycarbonyl group COOR ₉ at the end of the compound as a carbonyl group COOH. (D-3) is a reaction in which the amino group generated by (D-1) and the carboxyl group generated by (D-2) are condensed, and (D-1) and (D-2) The reaction of () may be performed first.

  Reaction (D-1) in the synthetic scheme of this invention performs the following reaction.

(In the reaction formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
Z represents an amino-protecting group different from Y.
W represents a protecting group for a hydroxy group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₈ represents a methyl group or an ethyl group.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; It represents a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom. )

In the above reaction formula, the compound represented by the general formula (6′-1-1) is a compound having a CO ₂ R ₈ group and an NHZ group at the terminal, and represented by the general formula (6′-1-2). The compound is a compound having a COOH group and an NHZ group at the terminal.
Further, the compound represented by the general formula (6′-2-1) is a compound having a CO ₂ R ₈ group and an amino group at the terminal, and the compound represented by the general formula (6′-2-2) is It is a compound having a carboxyl group and an amino group at the terminal.

Said reaction (D-1) is reaction which deprotects Z, and reaction conditions differ with the kind of Z.
For example, when Z is a benzyloxycarbonyl group, the benzyloxycarbonyl group can be deprotected in the presence of a palladium catalyst in a protic polar organic solvent.
Here, the protic polar solvent is not limited to these. For example, a mixed solvent of methanol / THF (tetrahydrofuran), a mixed solvent of ethanol / THF, and the like can be used.
Examples of the palladium catalyst include, but are not limited to, palladium / carbon, palladium hydroxide / carbon, and the like.

Further, when Z is a 9-fluorenylmethyloxycarbonyl group, it can be deprotected with a secondary amine.
When Z is a 2,2,2-trichloroethoxycarbonyl group, it can be deprotected by acting zinc powder-acetic acid or the like.
When Z is an allyloxycarbonyl group, it can be deprotected by adding an amine in the presence of a palladium catalyst.

  Reaction (D-2) in the synthetic scheme of this invention performs the following reaction.

(In the reaction formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
Z represents an amino-protecting group different from Y.
W represents a protecting group for a hydroxy group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₈ represents a methyl group or an ethyl group.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; It represents a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom. )

In the above reaction formula, the compound represented by the general formula (6′-1-1) is a compound having a CO ₂ R ₈ group and an NHZ group at the terminal, and represented by the general formula (6′-1-2). The compound is a compound having a COOH group and an NHZ group at the terminal.
Further, the compound represented by the general formula (6′-2-1) is a compound having a CO ₂ R ₈ group and an amino group at the terminal, and the compound represented by the general formula (6′-2-2) is It is a compound having a carboxyl group and an amino group at the terminal.

The above reaction (D-2) is a reaction in which the alkoxycarbonyl group COOR ₉ is converted to a carbonyl group COOH, and is performed by reacting with a strongly basic inorganic salt in a hydrous polar solvent.
Here, the hydrous polar solvent is not limited to these, but for example, a mixed solvent of THF and water, a mixed solvent of methanol and water, or the like can be used.
Moreover, as a strongly basic inorganic salt, although not necessarily limited to these, lithium hydroxide, sodium hydroxide, potassium hydroxide etc. can be used, for example.

  Reaction (D-3) in the synthetic scheme of this invention performs the following reaction.

(In the reaction formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
W represents a protecting group for a hydroxy group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; It represents a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom. )

The reaction (D-3) is a reaction in which the terminal amino group and carboxyl group of the compound represented by the general formula (6′-2-2) are condensed, and the presence of the phosphorus compound in the aprotic solvent Below, it can be reacted.
Here, the aprotic solvent is not limited to these, and for example, N, N-dimethylformamide, dichloromethane, tetrahydrofuran, dioxane, or a mixed solvent thereof can be used.
Examples of the phosphorus compound include diphenyl phosphate azide (DPPA), bis- (2-oxo-3-oxazolidinyl) phosphine acid chloride (BOPCl), and bis- (2-oxo-3-oxazolidinyl) phosphinic acid (BOP). Etc. can be used.

3-4. Synthesis of Macrocyclic Heptoxazole by Partial Cyclization of Macrocyclic Hexoxazole Next, macrocyclic heptoxazole is synthesized by cyclizing a part of the macrocyclic hexaoxazole by reactions (E) to (G). .
First, the reaction (E) in the synthesis scheme of the present invention performs the following reaction.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; It represents a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom. )

Said reaction (E) is reaction which remove | eliminates the protective group W of the hydroxyl group which the compound shown by General formula (7) has, The reaction conditions differ with kinds of W.
For example, when W is a tert-butyldimethylsilyl group, the tert-butyldimethylsilyl group can be eliminated by adding hydrogen fluoride in an aprotic solvent.
Further, when W is a p-methoxybenzyl group, it can be eliminated by hydrogenation reaction using palladium as a catalyst, Birch reduction or the like.
When W is a tert-butyl group, it can be deprotected under strongly acidic conditions such as trifluoroacetic acid or hydrochloric acid-ethyl acetate solution.
When W is an acetyl group, it can be deprotected with potassium carbonate in methanol.

When W is a benzoyl group, it can be deprotected under strong base conditions or strong hydride reduction conditions.
When W is a trimethylsilyl group, a triethylsilyl group, a triisopropylsilyl group, or a tert-butyldiphenylsilyl group, it can be deprotected by applying acidic conditions or fluoride ions.

  The reaction (F) in the synthesis scheme of the present invention performs the following reaction.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; An acyl group having 1 to 6 carbon atoms which may have an amino group or a guanidino group, a phenyl group which may have an amino group or guanidino group having a protecting group, a heterocyclic group having 1 to 6 carbon atoms, or Represents a halogen atom. )

In the reaction (F), the compound represented by the general formula (8) is reacted with methanesulfonyl chloride (MsCl) and triethylamine (Et ₃ N) in dichloromethane, and further diazabicycloundecene (DBU). The compound represented by the general formula (9) can be obtained by adding dehydration to intramolecular dehydration.

  The reaction (G) in the synthesis scheme of the present invention performs the following reaction.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; An acyl group having 1 to 6 carbon atoms which may have an amino group or a guanidino group, a phenyl group which may have an amino group or guanidino group having a protecting group, a heterocyclic group having 1 to 6 carbon atoms, or Represents a halogen atom. )

The reaction (G) of, in a polar aprotic solvent, N- bromosuccinimide (NBS) and cesium carbonate _(Cs 2 CO ₃₎ can be carried out by the action of.
Here, examples of the aprotic polar solvent include, but are not limited to, dichloromethane, tetrahydrofuran, acetonitrile, and the like.

  In addition, said reaction (G) is considered to be reaction through the following intermediate body (11).

Finally, by removing the protecting group Y of the amino group and the protecting groups contained in R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′, the compound of the present invention can be removed. can get. That is, reaction (H) which is the last step performs the following reaction.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently represent a hydrogen atom, an amino group having a protecting group, or a carbon number which may have a guanidino group. Having an alkyl group having 1 to 12, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or an aralkyl group having 7 to 12 carbon atoms which may have a guanidino group; It represents a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms, or a halogen atom. )

The reaction (H) is a reaction for removing the protective group Y and the protective group contained in R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′. Reaction conditions differ depending on the type of Y.
For example, when Y is a tert-butoxycarbonyl group, the tert-butoxycarbonyl group can be eliminated in an acidic solvent obtained by adding an acid such as hydrochloric acid or acetic acid to methanol, ethanol, dichloromethane, chloroform or the like. .
When Y is a 9-fluorenylmethyloxycarbonyl group, it can be deprotected with a secondary amine.
When Y is a 2,2,2-trichloroethoxycarbonyl group, it can be deprotected by acting zinc powder-acetic acid or the like.
When Y is an allyloxycarbonyl group, it can be deprotected by adding an amine in the presence of a palladium catalyst.

In addition, when the protecting group in R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ is the same functional group as the protecting group Y for the amino group, the above reaction (H) Can be finished with a single reaction. On the other hand, when the protecting groups for the amino groups Y, R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ are two or more types of functional groups, The reaction (H) can be completed by a plurality of reactions depending on the type of protecting group used.

3-5. Method for Synthesizing Trioxazole In the above-described synthesis scheme of the present invention, the compounds represented by the general formulas (4) and (5) are disclosed in Patent Document 2 (WO 02/48153) and Non-Patent Document 9 (Suzanne G. Rzuczek et al ., 2008, Bioorganic & Medicinal Chemistry Letters, vol.18, pp.913-917), Non-Patent Document 10 (Gurpreet Singh Minhas et al., 2006, Bioorganic & Medicinal Chemistry Letters, vol.16, pp.3891-3895) ), Non-Patent Document 11 (Masayuki Tera et al., 2006, Heterocycles, vol. 69, pp.505-514), Non-Patent Document 12 (Nobuaki Endoh et al, 2003, Heterocycles, vol. 60, pp. 1567) -1572), Sang-Ku Yoo et al., 1992, Tetrahedron letters, vol. 33, No. 16, pp. 2159-2162, etc., or by these improved methods.

  For example, the compound represented by the general formula (4) can be synthesized according to the following scheme using an amino acid derivative as a raw material.

(In the above formula, Z represents an amino-protecting group.
W represents a protecting group for a hydroxy group.
R ₈ represents a methyl group or an ethyl group.
R ₅ ′, R ₆ ′ and R ₇ ′ may each independently have a hydrogen atom, an amino group protected with Y (an amino group protecting group different from Z) or a guanidino group having a protecting group. A C1-C12 alkyl group, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group may have 7 to 12 carbon atoms. An aralkyl group, a phenyl group optionally having an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom is represented. )

The reaction (A-1) is a reaction in which a peptide bond is formed between an amino acid and an amino acid derivative to obtain a dipeptide. For example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCl), 1 -Hydroxybenzotriazole (HOBt) and triethylamine (Et ₃ N) can be used as reaction reagents.
The reaction (A-2) is a reaction in which the dipeptide obtained by the reaction (A-1) is closed to form an oxazole ring. This reaction can be performed in two steps, for example, a reaction using diethylaminosulfur trifluoride (DAST) as a reagent, followed by a reaction using diazabicycloundecene (DBU) and bromotrichloromethane (BrCCl ₃ ) as reagents. it can.
The reaction (A-3) is a reaction for eliminating the methyl group of the compound obtained in the reaction (A-2). This reaction can be performed, for example, by adding lithium hydroxide in a mixed solvent of tetrahydrofuran and water.
In this way, the compound represented by the general formula (12) can be obtained.

The reaction (A-4) is a reaction in which a peptide bond is formed between two amino acid derivatives to obtain a dipeptide. For example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCl), 1 -Hydroxybenzotriazole (HOBt) and triethylamine (Et ₃ N) can be used as reaction reagents.
The reaction (A-5) is a reaction in which the dipeptide obtained by the reaction (A-4) is closed to form an oxazole ring. This reaction can be performed in two steps, for example, a reaction using diethylaminosulfur trifluoride (DAST) as a reagent, followed by a reaction using diazabicycloundecene (DBU) and bromotrichloromethane (BrCCl ₃ ) as reagents. it can.
The reaction (A-6) is a reaction for deprotecting the protecting group Z of the amino group, and can be performed by the method already described in this specification.
In this way, the compound represented by the general formula (13) can be obtained.

The reaction (A-7) is a reaction in which a peptide bond is formed between the compound represented by the general formula (12) and the compound represented by the general formula (13) to obtain a tetrapeptide. -3- (3-Dimethylaminopropyl) carbodiimide (EDCl) and 4-dimethylaminopyridine (DMAP) can be reacted as reagents.
The reaction (A-8) is a reaction in which the tetrapeptide obtained by the reaction (A-7) is closed to form an oxazole ring. This reaction can be performed in two steps, for example, a reaction using diethylaminosulfur trifluoride (DAST) as a reagent, followed by a reaction using diazabicycloundecene (DBU) and bromotrichloromethane (BrCCl ₃ ) as reagents. it can.
In this way, the compound represented by the general formula (4) can be obtained.

  The compound represented by the general formula (5) can be synthesized according to the following scheme using an amino acid derivative as a raw material.

(In the above formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, Represents a cyclohexadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
Z represents an amino-protecting group different from Y.
W represents a protecting group for a hydroxy group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₉ represents a methyl group or an ethyl group.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )

The reaction (B-1) is a reaction in which a peptide bond is formed between two amino acid derivatives to obtain a dipeptide. For example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCl), 1 -Hydroxybenzotriazole (HOBt) and triethylamine (Et ₃ N) can be used as reaction reagents.
The reaction (B-2) is a reaction in which the dipeptide obtained by the reaction (B-1) is closed to form an oxazole ring. This reaction can be performed in two steps, for example, a reaction using diethylaminosulfur trifluoride (DAST) as a reagent, followed by a reaction using diazabicycloundecene (DBU) and bromotrichloromethane (BrCCl ₃ ) as reagents. it can.
The reaction (B-3) is a reaction for eliminating the methyl group of the compound obtained in the reaction (B-2). This reaction can be performed, for example, by adding lithium hydroxide in a mixed solvent of tetrahydrofuran and water.
In this way, the compound represented by the general formula (14) can be obtained.

The reaction (B-4) is a reaction in which a peptide bond is formed between two amino acid derivatives to obtain a dipeptide. For example, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDCl), 1 -Hydroxybenzotriazole (HOBt) and triethylamine (Et ₃ N) can be used as reaction reagents.
The reaction (B-5) is a reaction in which the dipeptide obtained by the reaction (B-4) is closed to form an oxazole ring. This reaction can be performed in two steps, for example, a reaction using diethylaminosulfur trifluoride (DAST) as a reagent, followed by a reaction using diazabicycloundecene (DBU) and bromotrichloromethane (BrCCl ₃ ) as reagents. it can.
The reaction (B-6) is a reaction for deprotecting the protecting group Z of the amino group and can be carried out by the method already described in this specification.
In this way, the compound represented by the general formula (15) can be obtained.

The reaction (B-7) is a reaction in which a peptide bond is formed between the compound represented by the general formula (14) and the compound represented by the general formula (15) to obtain a tetrapeptide. -3- (3-Dimethylaminopropyl) carbodiimide (EDCl) and 4-dimethylaminopyridine (DMAP) can be reacted as reagents.
The reaction (B-8) is a reaction in which the tetrapeptide obtained by the reaction (B-7) is closed to form an oxazole ring. This reaction can be performed in two steps, for example, a reaction using diethylaminosulfur trifluoride (DAST) as a reagent, followed by a reaction using diazabicycloundecene (DBU) and bromotrichloromethane (BrCCl ₃ ) as reagents. it can.
In this way, the compound represented by the general formula (5) can be obtained.

3-6. Amino Acid Derivatives as Raw Materials As raw materials for producing the compound of the present invention, that is, amino acid derivatives used in the above reactions (A-1), (A-4), (B-1) and (B-4), Derivatives obtained from known amino acids as raw materials can be used, and derivatives obtained from newly synthesized novel amino acids as raw materials can also be used.

Here, examples of known amino acids include, but are not limited to, ornithine, 2,5-diaminopentanoic acid, lysine, 2,6-diaminocaproic acid, arginine, 2-amino-5-guanidino. pentanoic acid, homoarginine ^{(N 6} - (aminoiminomethyl) -L- lysine), 2,7-diamino-7-Iminoheputan acid, Arubijiin (3- (aminocarbonylamino) -L- alanine), citrulline, homocitrulline ^{(N 6} - (aminocarbonyl) -L- lysine), asparagine, 2-amino-3-carbamoyl propanoic acid, glutamine, 2-amino-4-carbamoyl-butyric acid, Gigaruchinin ^(N 5 - ^{[[(aminoiminomethyl)} amino ] Carbonyl] -L-ornithine), aminocremycin (2-amino-4- (4-a -2,5-cyclohexadienyl) butyric acid), tetrahydrolatyrin (α, 2-diamino-1,4,5,6-tetrahydro-4-pyrimidinepropanoic acid), serine, threonine, allothreonine, β-hydroxy Norvaline (3-hydroxy-L-norvaline), 2-amino-3-hydroxypentanoic acid, 3-hydroxyleucine, 2-amino-3-hydroxy-4-methylpentanoic acid, 3-hydroxyornithine, 3-hydroxylysine 3-phenylserine, 2-amino-3-hydroxy-3-phenylpropanoic acid, 3- (2-thienyl) serine, α-amino-β-hydroxy-2-thiophenepropionic acid, α-amino-β-hydroxy -2-furanpropionic acid and the like can be used.

  Derivatives of amino acids can be obtained, for example, by using the above known amino acids as alkyl esters or by adding a protecting group. Here, the alkyl ester is a methyl ester or an ethyl ester. The alkyl ester can be obtained, for example, by reacting an amino acid with an alcohol, but may be a derivative obtained by another method. Further, as the protecting group, the same protecting group as that of Y, Z or W described above can be used, and other protecting groups can be used.

Here, when an amino acid is protected with a tert-butoxycarbonyl group or a benzyloxycarbonyl group which is a protective group for an amino group, for example, but not limited thereto, in the presence of a base, di- It can be protected by reacting a tert-butoxycarbonyl group or benzyl chloroformate.
In the case of protecting a hydroxyl group with a tert-butyldimethylsilyl group, p-methoxybenzyl group or the like which is a protecting group for a hydroxyl group, for example, but not limited thereto, in the presence of a base, Protection can be achieved by reacting tert-butyldimethylsilyl chloride or p-methoxybenzyl chloride.

The compound of this invention can obtain the compound which has a desired structure by selecting the amino acid which is a raw material suitably. For example, in order to produce a compound in which X in the general formula (1) is —CH ₂ —, the amino acid derivative used in the above reaction formula (B-1) is not limited thereto, but ornithine, A derivative such as 2,5-diaminopentanoic acid, lysine or 2,6-diaminocaproic acid (with a protecting group added) may be used.
Moreover, in order to produce a compound in which X in the general formula (1) is —NHC (NH) —, the amino acid derivative used in the above reaction formula (B-1) is not limited to these, For example, derivatives such as arginine, 2-amino-5-guanidinopentanoic acid, and homoarginine (N ^6- (aminoiminomethyl) -L-lysine) may be used.
Next, in order to produce a compound in which X in the general formula (1) is —CH (NH ₂ ) —, the amino acid derivative used in the above reaction formula (B-1) is not limited to these. However, a derivative such as 2,5,5-triaminopentanoic acid may be used.

In order to produce a compound in which X in the general formula (1) is —C (NH) —, the amino acid derivative used in the above reaction formula (B-1) is not limited to these. A derivative of 2,7-diamino-7-iminoheptanoic acid may be used.
In order to produce a compound in which X in the general formula (1) is —NHCO—, the amino acid derivative used in the above reaction formula (B-1) is not limited to these, but for example, albidiyne (3 - (aminocarbonylamino) -L- alanine), citrulline or homocitrulline - may be used ^{(N 6} (aminocarbonyl) derivative -L- lysine).
In order to produce a compound in which X in the general formula (1) is —CO—, the amino acid derivatives used in the above reaction formula (B-1) are not limited to these. For example, asparagine, 2 A derivative of -amino-3-carbamoylpropanoic acid, glutamine or 2-amino-4-carbamoylbutyric acid may be used.

In order to produce a compound in which X in the general formula (1) is —NHCONHC (NH) —, the amino acid derivative used in the above reaction formula (B-1) is not limited to these. Gigaruchinin - it may be used ^(N 5 ^{[[(Aminoiminomethyl)} amino] carbonyl] -L- ornithine) derivatives and the like.
In order to produce a compound in which X in the general formula (1) is a cyclohexadienylene group, amino acid derivatives used in the above reaction formula (B-1) are not limited to these. Derivatives such as cremycin (2-amino-4- (4-amino-2,5-cyclohexadienyl) butyric acid may be used.
In order to produce a compound in which X in the general formula (1) is a 3,4,5,6-tetrahydropyrimidin-2,6-yl group, as amino acids used in the above reaction formula (B-1), Although not limited thereto, for example, a derivative such as tetrahydroratillin (α, 2-diamino-1,4,5,6-tetrahydro-4-pyrimidinepropanoic acid) may be used.

R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the general formula (1) can have a desired structure by appropriately selecting the amino acid as a raw material.
For example, in order to make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ into hydrogen atoms, the above reactions (A-1), (A-4), (B-1) and ( As the amino acid derivative used in B-4), a serine derivative (an alkyl ester or a protecting group added) may be used.
In general _{R 2,} R ₃ in the formula _{(1), R 4, R} 5, R 6 and _{R 7,} for the amino group or an alkyl group having 1 to 12 carbon atoms which may have a guanidino group Examples of the amino acid derivatives used in the above reactions (A-1), (A-4), (B-1) and (B-4) include, but are not limited to, for example, threonine, allothreonine , Β-hydroxynorvaline (3-hydroxy-L-norvaline), 2-amino-3-hydroxypentanoic acid, 3-hydroxyleucine, 2-amino-3-hydroxy-4-methylpentanoic acid, 3-hydroxyornithine or A derivative such as 3-hydroxylysine may be used.

In order to make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the general formula (1) an amino group, the above reactions (A-1), (A-4), (B— As the amino acid derivative used in 1) and (B-4), for example, a derivative such as 2,3-diamino-3-hydroxypropionic acid may be used.
In order to make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the general formula (1) into a guanidino group, the above reactions (A-1), (A-4), (B— As the amino acid derivative used in 1) and (B-4), for example, a derivative such as 2-amino-3-guanidino-3-hydroxypropionic acid may be used.
To make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the general formula (1) an aralkyl group having 7 to 12 carbon atoms which may have an amino group or a guanidino group The amino acid derivatives used in the above reactions (A-1), (A-4), (B-1) and (B-4) are not limited to these. For example, 2-amino-3- Derivatives such as hydroxy-3-benzylpropanoic acid may be used.

In order to make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in general formula (1) a phenyl group optionally having an amino group or a guanidino group, the above reaction (A- The amino acid derivatives used in 1), (A-4), (B-1) and (B-4) are not limited to these, but include, for example, 3-phenylserine or 2-amino-3-hydroxy A derivative such as -3-phenylpropanoic acid may be used.
In order to make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the general formula (1) into a heterocyclic group having 2 to 6 carbon atoms, the above reactions (A-1), (A -4), amino acid derivatives used in (B-1) and (B-4) are not limited to these, but include, for example, 3- (2-thienyl) serine, α-amino-β-hydroxy- Derivatives such as 2-thiophenepropionic acid and α-amino-β-hydroxy-2-furanpropionic acid may be used.
In order to make R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ in the general formula (1) into halogen atoms, the above reactions (A-1), (A-4), (B— The amino acid derivatives used in 1) and (B-4) are not limited to these, but for example, derivatives such as 2-amino-3-chloro-3-hydroxypropionic acid may be used.

3-7. Conversion of Substituent The compound of the present invention can be obtained in a desired structure by appropriately selecting the amino acid used as a raw material as described above. However, it is also possible to obtain a compound having a desired structure by converting a substituent of a compound once synthesized into another substituent.
For example, when the side chain of the compound of the present invention is an aminoalkyl group, it can be converted to a ureidoalkyl group as shown in the following reaction.

  In the above reaction, a macrocyclic heptaoxazole having an aminoalkyl group is converted into mercuric chloride and 1,3-bis (tert-butoxycarbonyl) -2-methyl-2-thiopseurea urea (1,3-Bis ( By adding tert-butoxycarbonyl) -2-methyl-2-thiopseudorea) and then reacting in a solvent to which trifluoroacetic acid is added, a macrocyclic heptaoxazole having a ureidoalkyl group can be produced.

3-8. As described above for the intermediate compound, in the scheme for synthesizing the compound of the present invention, the general formula (4) and the general formula (5) are used as starting compounds, and the general formulas (6) to (10) are used as intermediate compounds. In general, a compound of the general formula (1) is synthesized. Here, the intermediate compounds of general formulas (6) to (10) are all useful as compounds for synthesizing the compound of general formula (1). In particular, the compound represented by the general formula (10) has a protecting group included in the protecting groups Y, R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ against the amino group. Except for this, it has the same chemical structure as the general formula (1). Therefore, the compound represented by the general formula (10) is a particularly useful compound for producing the compound of the present invention.

4). Low molecular probe for visualizing the G-quadruplex structure The compound of the present invention exhibits an activity of strongly binding to the G-quadruplex structure. Therefore, by linking the compound of the present invention and a fluorescent compound, a low molecular probe that visualizes the presence of a G-quadruplex structure can be obtained. Here, the fluorescent compound can be linked to the compound of the present invention by using a fluorescent compound capable of reacting with the side chain amino group in the compound of the present invention.

Here, as the fluorescent compound that can react with an amino group, a compound in which a functional group that reacts with an amino group is added to a fluorescent dye molecule can be used. Here, the fluorescent dye molecule is not limited to these, for example, BODIPY (trade name, Molecular Probe), fluorescein, TAMRA, 5-FAM, 6-FAM, Alexa Fluor (trade name, Molecular Probe Co., Ltd.) and Cascade Blue (trademark, Molecular Probe Co., Ltd.) can be used. And, to these fluorescent dye molecules, via a linker as necessary, such as succinimidyl ester (succinimidyloxycarbonyl group), acid halide (halogenoxycarbonyl group), isothiocyanate group, carboxyl group, etc. By adding a functional group capable of reacting with an amino group, a target fluorescent compound can be obtained.
In addition, as such a fluorescent dye molecule to which a functional group capable of reacting with an amino group is added, a commercially available one may be used, for example, FITC (Dojin Chemical Co., Ltd.), BODIPY-TRX SE (Molecular Probe Co., Ltd.). ), 5 (6) -TAMRA-X SE (ANA SPEC) or the like can be used.

  As an example, when a fluorescent dye molecule to which a succinimidyl ester (succinimidyloxycarbonyl group) is added is used as the fluorescent compound, the compound represented by the general formula (1) as shown in the following reaction formula: The probe of the present invention can be obtained by reacting with a fluorescent dye molecule to which a succinimidyl ester represented by the general formula (16) is added.

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadienylene group and a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Q represents a fluorescent dye.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom.
R ₁₀ represents an alkylene group having 1 to 12 carbon atoms. )

Although the above reaction can be carried out in a basic solvent, it is particularly preferred to carry out in triethylamine. Moreover, reaction temperature is about 20-80 degreeC normally.
In the general formula (16), Q represents a fluorescent dye, but the fluorescent dye is not particularly limited. For example, BODIPY (trade name, Molecular Probe), fluorescein, TAMRA, 5-FAM, 6-FAM, Alexa Fluor (trade name, Molecular Probe) and Cascade Blue (trade name, Molecular Probe) can be used.
In the general formula (16), R ₁₀ represents an alkylene group having 1 to 12 carbon atoms, but includes not only a linear group but also an alkylene group having a branched chain.

Since the probe of the present invention binds strongly to the G-quadruplex structure, the G-quadruplex structure can be measured by detecting fluorescence derived from the fluorescent compound. For example, by contacting the nucleic acid to be measured and the probe of the present invention (for example, mixing in a solution), after removing the unreacted probe, detecting fluorescence derived from the fluorescent compound in the nucleic acid, It can be determined whether or not the nucleic acid contains a G-quadruplex structure. In this case, the presence of the G-quadruplex structure contained in the nucleic acid to be measured can be quantitatively determined based on the fluorescence intensity.
In addition, the probe of the present invention can be incorporated into an isolated living cell or tissue and detect the fluorescence derived from the fluorescent compound, thereby measuring the position of the G-quadruplex structure in the living cell or tissue. Can do. Even in this case, the abundance can be determined quantitatively together with the location of the G-quadruplex structure contained in the measurement target living cell or tissue based on the fluorescence intensity.

  EXAMPLES Next, although an Example is shown and this invention is demonstrated further in detail, the technical scope of this invention is not limited to these Examples.

Example 1: Synthesis of compounds of the present invention
L1H1-7OTD having a macrocyclic structure composed of seven oxazoles and having an aminobutyl group as a side chain was synthesized according to the following scheme 1.

(1) Reaction a
Trioxazole 4 was dissolved in a mixed solvent of tetrahydrofuran (THF) and methanol aerated with hydrogen, and the benzyloxycarbonyl group (Cbz) was eliminated in the presence of palladium hydroxide / carbon.
(2) Reaction b
A tricarboxylic acid was produced by dissolving trioxazole 5 in a mixed solvent of THF and water and adding lithium monohydrate.
(3) Reaction c
An amine synthesized from trioxazole 4 and a carboxylic acid synthesized from triazole 5 are dissolved in a mixed solvent of THF and water, and N-methylmorpholine 4- (4,6-dimethoxy-1,3,5-triazine-2 is obtained. -Il) -4-methylmorpholinium chloride (DMT-MM) and N-methylmorpholine (NMM) were added to condense the amine and carboxylic acid. Linear hexaoxazole 6 was obtained from trioxazole 4 and trioxazole 5 in a yield of 91%.
(4) Macrocyclization of hexaoxazole Under the same conditions as in reaction a, the Cbz group of hexaoxazole 6 was eliminated, and the methyl ester of hexaoxazole 6 was deprotected under the same conditions as in reaction b. Thereafter, the resulting amino acid was dissolved in a mixed solvent of N′N-dimethylformamide and dichloromethane to which diisopropylethylamine (Et ⁱ Pr ₂ N), 4-dimethylaminopyridine (DMAP) and diphenylphosphoric acid azide (DPPA) were added. Cyclization was performed by highly diluting to 3 mM to obtain macrocyclic bisamide from hexaoxazole 6 in a yield of 57%.

(5) Reaction e
Bisamide 7 was dissolved in THF solvent, and tert-butyldimethylsilyl (TBS) group was deprotected with HF · pyridine to obtain alcohol 8.
(6) Reaction f
Alcohol 8 was dissolved in dichloromethane, methanesulfonyl chloride (MsCl) and triethylamine (Et ₃ N) were added, converted to olefin 9 by mesylation, and treated with diazabicycloundecene (DBU).

(7) Reaction g
Olefin 9 was reacted with N-bromosuccinimide (NBS) in acetonitrile solvent and the resulting bromide was treated with triethylamine (Et ₃ N) and further cyclized with cesium carbonate to give heptaoxazole 11.
(8) Reaction h
Finally, heptaoxazole 11 was dissolved in chloroform and the tert-butoxycarbonyl (Boc) group was deprotected with trifluoroacetic acid (TFA) to obtain L1H1-7OTD from olefin 9 in a yield of 30%.

(9) Measurement of NMR spectrum About L1H1-7OTD synthesize | combined by the said scheme, it measured with the nuclear magnetic resonance apparatus in the deuterium substitution solvent, and measured the NMR spectrum. The results are as follows.
¹ H NMR (400 MHz, ref 2.5 ppm for DMSO d-6)
δ8.97 (s, 1H), 8.93 (s, 1H), 8.86 (s, 1H), 8.83 (s, 2H), 8.71 (s, 1H), 8.66 (s, 1H), 8.06 (br, 2H) , 7.62 (br, 1H), 5.33 (br, 1H), 2.06 (m, 1H), 1.70 (br, 3H), 1.24 (br, 1H), 0.908 (br, 1H).
¹³ C NMR (100 MHz, ref 49.0 ppm for DMSO d-6)
δ172.6, 168.1, 165.2, 164.7, 164.6, 163.7, 152.3, 152.1, 150.4, 150.3, 150.1, 149.9, 149.4, 149.2, 148.6, 148.5, 148.2, 148.0, 145.2, 139.0, 138.3, 138.2, 138.1, 137.4, 57.27, 57.10, 43.46, 41.96, 36.00, 28.98.
(10) Mass spectrometry The L1H1-7OTD synthesized by the above scheme was measured with an ESI-MS apparatus and the molecular weight was measured. The molecular weight was 598.1467. It was also was estimated elemental composition from the resulting molecular weight value _{C 27 H 19 N 9 O 8} Na. From these results, it was confirmed that the target compound could be synthesized.
(11) Measurement of optical rotation The L1H1-7OTD synthesized by the above scheme was dissolved in a mixed solvent of chloroform and methanol, and measurement was performed using an optical rotation photometer. The obtained value was [α] ^D = 77.1, and it was confirmed that a compound with high optical purity was obtained.

( Example 2: Measurement of telomerase inhibitory activity )
The synthesized compound L1H1-7OTD of the present invention was tested for telomerase inhibitory activity using the TRAP assay. The TRAP assay is widely used to conveniently evaluate the telomerase inhibitory activity of G-quadruplex binding agents. This assay showed a very potent telomerase inhibitory activity with an IC ₅₀ of 0.8 nM. This compound is the most potent telomerase inhibitor reported to date.
The TRAP assay is an inhibitor against telomerase that is crudely purified from cell lysates of human B lymphoma Namalwa cells according to the protocol described in Tabata, Y. et al., J. Antibiot., 1999, vol. 52, pp. 412. Activity was measured.

(Example 3: Measurement of interaction of L1H1-7OTD with telomeres)
The selective interaction between the telomere DNA sequence and the compound L1H1-7OTD of the present invention was tested by PCR stop assay using telo24 and its mutant sequence telo24-mut. According to the PCR protocol, the inhibitory activity of L1H1-7OTD on the chain extension reaction of telo24 and telo24-mut was evaluated. By this test, L1H1-7OTD strongly inhibited telo24 PCR with an IC ₅₀ of 0.67 ± 0.01 μM. On the other hand, the inhibitory activity of L1H1-7OTD against telo24-mut was weak, and its IC ₅₀ value was 5.2 ± 0.8 μM. From these results, it was shown that L1H1-7OTD interacts selectively with telo24 more strongly by 7.7 times or more than the mutant sequence of telo24.

( Comparative Example 1: Compound having six oxazoles in macrocyclic structure )
(1) Synthesis of L2H2-6OTD As a comparative example, a compound (L2H2-6OTD) having a side chain having an amino group or a guanidino group but having six oxazoles constituting a macrocyclic structure was synthesized according to the following scheme. .

(1-1) Reaction a
Trioxazole 3 was dissolved in a mixed solvent of THF and water, and lithium hydroxide was added to generate carboxylic acid.
(1-2) Reaction b
Trioxazole 3 was dissolved in a mixed solvent of tetrahydrofuran (THF) and methanol aerated with hydrogen, and the benzyloxycarbonyl (Cbz) group was deprotected in the presence of palladium hydroxide / carbon to form an amine. .
(1-3) Reaction c
Carboxylic acid 4 produced by reaction a and amine 5 produced by reaction b are combined with N-methylmorpholine 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4- Methyl morpholinium chloride (DMT-MM) and N-methylmorpholine (NMM) were added for condensation to give linear hexaoxazole 6.
(1-4) Reaction d
Hexoxazole 6 was dissolved in a mixed solvent of THF and water, and lithium hydroxide was added to convert the terminal methoxycarbonyl group into a carboxyl group.
(1-5) Reaction e
In hexaxazole 6 in a mixed solvent of THF and methanol, the terminal benzyloxycarbonyl (Cbz) group was deprotected to form an amino group in the presence of palladium hydroxide / carbon.

(1-6) Reaction f
Diisopropylethylamine (Et ⁱ Pr ₂ N), 4-dimethylaminopyridine (DMAP) and bis (2-oxo-3-oxazolidinyl) phosphinic chloride (BOPCl) in a mixed solvent of N, N-dimethylformamide and dichloromethane Was added, and the carboxyl group of hexazole 6 produced by reaction d and the amino group of hexaoxazole 6 produced by reaction e were condensed to form macrocyclic hexaoxazole.
(1-7) Reaction g
The tert-butoxycarbonyl (Boc) group, which is a protecting group for the two side chains R of macrocyclic hexaoxazole, is deprotected by adding trifluoroacetic acid in dichloromethane to acidify the diaminobutyl macrocyclic hexaoxazole 2b. Got.
(1-8) Reactions h and i
Add triethylamine, mercury chloride and 1,3-bis (tert-butoxycarbonyl) -2-methyl-2-thiopseudorea (1,3-Bis (tert-butoxycarbonyl) -2-methyl-2-thiopseudorea), then The amino group of diaminobutyl macrocyclic hexaoxazole 2b was converted to a guanidino group in dichloromethane with trifluoroacetic acid added to diguanidinobutyl macrocyclic hexaoxazole 2c.
(1-9) Reaction i
By reacting with acetic anhydride in a dichloromethane solvent, the guanidino group in the side chain of diguanidinobutyl macrocyclic hexaoxazole 2c was converted to acetylamine.

(2) Measurement of telomerase inhibitory activity The telomerase inhibitory activity of L2H2-6OTD synthesized by the above scheme was measured by the TRAP assay described in Example 2.
As a result, the telomerase inhibitory activity of L2H2-6OTD is as follows. For macrocyclic hexaoxazole (2b) whose side chain end is an amino group, the IC ₅₀ is 20 nM and the side chain end is a guanidino group. For (2c), the IC ₅₀ was 20 nM. This was considerably lower than the telomerase inhibitory activity (IC ₅₀ = 0.8 nM) of the compound of the present invention synthesized in Example 1.

( Comparative Example 2: Compound having no amino acid side chain )
(1) Synthesis of S1T1-7OTD As a next comparative example, the following compound (S1T1-7OTD) having a macrocyclic structure composed of seven oxazoles but having no side chain with an amino group was prepared as follows. It was synthesized according to the scheme described in Dairin Ishizuka et al., 5 others, published on March 12, 2008, Proceedings of the Chemical Society of Japan, Vol.88, No.2, pp.1049).

(2) Measurement of telomerase inhibitory activity The telomerase inhibitory activity of S1T1-7OTD synthesized according to the above scheme was measured by the TRAP assay described in Example 2.
As a result, the telomerase inhibitory activity of S1T1-7OTD was IC ₅₀ = 10 nM. This was considerably lower than the telomerase inhibitory activity (IC ₅₀ = 0.8 nM) of the compound of the present invention synthesized in Example 1.
The results of measuring the telomerase inhibitory activity of the compounds of the present invention and the compounds synthesized in Comparative Examples and telomestatin are shown in the following table.

( Example 3: Measurement of antitumor activity )
For the measurement of antitumor activity, HeLa cells (cervical cancer cell lines), which are cancer cells that overexpress telomerase, and Saos-2 cells (osteoblastoma, which are cancer cells that do not express telomerase). Cell lines). Moreover, as an antitumor agent, the compound L1H1-7OTD of this invention synthesize | combined in Example 1 was used, and the representative antitumor agent doxorubicin (doxorubicin) was used as a comparison. Doxorubicin is an antitumor agent that suppresses tumor growth by being inserted between DNA base pairs and inhibiting DNA amplification, and does not inhibit tumor growth by inhibiting telomerase.
For the culture of the two cancer cells, modified Dulbecco's Eagle's medium containing 10% fetal bovine serum, 50 μg / ml streptomycin and 5 U / ml penicillin was used. 2 × 10 ³ cancer cells per well were seeded in a 96-well plate and treated with various concentrations of antitumor agents (L1H1-7OTD and doxorubicin) for 6 days. Cancer cell viability was measured by MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay. In the 6 days of culture, the concentration of the anti-tumor agent viability of cancer cells is 50% and the IC _50.

As a result, L1H1-7OTD was a cancer cell that showed cytotoxicity of 2.2 ± 0.5 μM in IC ₅₀ against HeLa cells, which are cancer cells that overexpress telomerase, but does not express telomerase. It was not cytotoxic to Saos-2 cells. Meanwhile, doxorubicin exhibits antitumor activity without depending on telomerase, IC ₅₀ against HeLa cells indicates the cytotoxicity of 0.03 ± 0.001 .mu.M, cytotoxicity IC ₅₀ of 0.07 ± 0.005 relative to Saos-2 cells showed that.
From these results, the compound of the present invention exerts an antitumor activity by specifically acting on cancer cells overexpressing telomerase, and does not act on cells not expressing telomerase. It became clear.

( Example 4: Synthesis of macrocyclic heptaoxazole having a ureidobutyl group )
L1H1-7OTD synthesized in Example 1 was dissolved in triethylamine with mercury chloride and 1,3-bis (tert-butoxycarbonyl) -2-methyl-2-thiopseurea (1,3-Bis (tert-butoxycarbonyl) -2-methyl-2-thiopseudorea) was added, and then reacted in a solvent to which trifluoroacetic acid was added to convert the aminobutyl group into a ureidobutyl group. The reaction formula is shown below.

( Example 5: Synthesis of low molecular probe for visualizing G-quadruplex structure )
L1H1-7OTD synthesized in Example 1 was reacted with propionic acid succinimide linked with BODIPY as a fluorescent dye in triethylamine. The reaction was performed at 60 ° C., and the product (L1BOD-7OTD) was obtained with a yield of 80%. The reaction formula is shown below.

( Example 6: Functional evaluation of probe )
Telo24 (5-TTA GGG TTA GGG TTA GGG TTAGGG-3: SEQ ID NO: 1) that forms a G-quadruplex structure and telo24-mut (single-stranded DNA that does not form a G-quadruplex structure) 5- TTA GAG TTA GAG TTA GAG TTA GGG-3: SEQ ID NO: 2) was prepared. Next, these were mixed with the L1H1-7OTD synthesized in Example 1 and the probe synthesized in Example 5 in eight combinations shown in the table of FIG. 1, and eight of 12% polyacrylamide gel were mixed. In the lane, electrophoresis was performed using TBE buffer. In FIG. 1, “+” indicates that 1 equivalent of the compound is added to the DNA, and “++” indicates that 10 equivalent of the compound is added to the DNA. The single-stranded DNA after electrophoresis was stained with Stains-all (manufactured by Sigma-Aldrich) and detected with the fluorescence wavelength of Stains-all and the fluorescence wavelength of BODIPY. In Fig. 1, “L1BOD-7OTD” is a photograph of the emission of the gel when excited with the fluorescence wavelength of BODIPY, and “Stains-all” is the emission of the gel when excited with the fluorescence wavelength of BODIPY. It is a photograph taken and “Merged” is a superposition of these two. From these results, it was shown that the probe of the present invention can detect only telo24 forming a G-quadruplex structure.

  Since the compound of the present invention has high telomerase inhibitory activity, it is useful as a biochemical reagent used for studying cancer mechanisms and animal and plant development mechanisms, and is also useful as an antitumor agent with few side effects.

SEQUENCE LISTING

<110> National University Corporation Tokyo University of Agriculture and Technology
National Institute of Advanced Industrial Science and Technology

<120> A telomerase inhibitor

<130> 090077

<150> JP 2008-166494
<151> 2008-06-25

<160> 2

<170> PatentIn version 3.4

<210> 1
<211> 24
<212> DNA
<213> Artificial

<220>
<223> Synthetic DNA

<400> 1
ttagggttag ggttagggtt aggg 24


<210> 2
<211> 24
<212> DNA
<213> Artificial

<220>
<223> Synthetic DNA

<400> 2
ttagagttag agttagagtt aggg 24

Claims (15)

A compound represented by the following general formula (1).

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom. ) X is —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, or —NHCONHC (NH) —. A compound according to claim 1, characterized. X _{is, -CH 2 -, - NHC (} NH) - or -CH (NH ₂₎ - characterized in that it is a compound according to claim 2. 4. The compound according to claim 3, wherein X is —CH ₂ —, R ₁ is a linear alkylene group having 1 to 7 carbon atoms, and is represented by the following general formula (2).

(In formula, n represents the integer of 2-8.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 2 to 6 carbon atoms or a halogen Represents an atom. )   Compound according to claim 4, characterized in that n is 4. R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently a hydrogen atom or an alkyl group having 1 to 7 carbon atoms which may have an amino group or a guanidino group. A compound according to any one of claims 1 to 5, characterized. The compound according to claim 6, wherein any of R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ is a hydrogen atom. 2. The compound according to claim 1, wherein X is —CH ₂ —, R ₁ is an alkylene group having 3 carbon atoms, and R ₂ , R ₃ , R ₄ , R ₅ , R _6, and R ₇ are hydrogen atoms. And a compound represented by the following structural formula (3):

  A salt of the compound according to claim 1.   A telomerase inhibitor comprising the compound according to any one of claims 1 to 8 or a salt thereof.   An antitumor agent comprising the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof as an active ingredient. A method for producing the compound according to claim 1, comprising the following steps (F) to (H):
(F) By reacting a compound represented by the following general formula (8) with methanesulfonyl chloride (MsCl) and triethylamine (Et ₃ N) in dichloromethane, and further adding diazabicycloundecene (DBU), A step of obtaining a compound represented by the following general formula (9) by intramolecular dehydration;

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )
(G) In an aprotic polar solvent, N-bromosuccinimide (NBS) and cesium carbonate (Cs ₂ CO ₃ ) are allowed to act on the compound represented by the following general formula (9) to obtain the following general formula ( A step of obtaining a compound shown in 10);

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )
(H) Protecting groups Y and R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ included in the compound represented by the following general formula (10) are removed. The process of making it leave | separate and obtaining the compound shown by following General formula (1).

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Y represents an amino-protecting group.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms or a halogen Represents an atom.
R ₂ ′, R ₃ ′, R ₄ ′, R ₅ ′, R ₆ ′ and R ₇ ′ each independently have a hydrogen atom, an amino group having a protecting group, or a guanidino group having a protecting group. May have an alkyl group having 1 to 12 carbon atoms, an amino group having a protecting group, a guanidino group having a protecting group, an amino group having a protecting group, or a guanidino group having a protecting group. A phenyl group optionally having an aralkyl group, an amino group having a protecting group or a guanidino group having a protecting group, a heterocyclic group having 2 to 6 carbon atoms, or a halogen atom. )   The production method according to claim 12, wherein the protective group Y is a tert-butoxycarbonyl group (Boc group), and the protective group Y (Boc group) is eliminated in an acidic solvent.   The probe which visualizes the G-quadruplex structure which made the compound in any one of Claims 1-8 react with the fluorescent compound which can react with the amino group in the said compound. The probe according to claim 14, wherein the fluorescent compound is a compound represented by the general formula (16), which is a compound represented by the general formula (17).

(In the formula, X represents —CH ₂ —, —NHC (NH) —, —CH (NH ₂ ) —, —C (NH) —, —NHCO—, —CO—, —NHCONHC (NH) —, cyclohexane, It represents a sadielenylene group or a 3,4,5,6-tetrahydropyrimidin-2,6-yl group.
Q represents a fluorescent dye.
R ₁ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms.
R ₂ , R ₃ , R ₄ , R ₅ , R ₆ and R ₇ are each independently an alkyl group having 1 to 12 carbon atoms, an amino group, which may have a hydrogen atom, an amino group or a guanidino group, A guanidino group, an amino group or a guanidino group optionally having an aralkyl group having 7 to 12 carbon atoms, a phenyl group optionally having an amino group or a guanidino group, a heterocyclic group having 1 to 6 carbon atoms or a halogen Represents an atom.
R ₁₀ represents a single bond, an alkylene group having 1 to 12 carbon atoms, an alkenylene group having 1 to 12 carbon atoms, or an alkynylene group having 1 to 12 carbon atoms. )

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