JP2013040112A - 11c labeled isoquinoline derivative, preparation method therefor, precursor thereof, probe for pet using the same and method for tissue imaging - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provideC labeled isoquinoline derivatives labeled with [C] methyl group on isoquinoline ring present in fasudil, analogs thereof which are Rho kinase inhibitors, and preparation method therefor and precursors suitably used in preparation of theC labeled isoquinoline derivatives.SOLUTION: There are exemplified, (S)-hexahydro-2-methyl-1-[(4-[C] methyl-5-isoquinolinyl)sulfonyl]-1H-1,4-diazepine, etc., as theC labeled isoquinoline derivatives. The compounds are obtained in a short time by using precursors having protective groups such as a trihaloacetyl group, after introducing a [C] methyl group onto the isoquinoline ring of the precursors, removing the protective group under a basic condition. The compounds can be suitably used as an imaging agent in imaging a tissue by using PET method, etc.

Description

The present invention relates to an ¹¹ C-labeled isoquinoline derivative having a [ ¹¹ C] methyl group introduced on the isoquinoline ring of isoquinoline sulfonamide compounds, a production method thereof, a precursor thereof, a molecular probe, and a tissue imaging method.

 Positron emission tomography (PET) is the only technique that can non-invasively image in vivo molecules from small animals to humans. For this reason, in recent years, expectations for the use of the PET method in the medical and drug discovery fields are rapidly expanding. However, the synthesis of a PET probe, which is the key to technology utilization, is difficult because the lifetime of the radiation nucleus incorporated in the molecule is short.

Conventionally, only [ ¹¹ C] methylation on heteroatoms such as O (oxygen), N (nitrogen), and S (sulfur) has been performed because of the easy introduction. However, in the living body, these heteroatoms are used as clues, and they are easily metabolized by the enzyme, and the chemical bond at the labeled site is cleaved by metabolism, so there is a problem that the authenticity of the obtained PET image remains a question. It has been.

In order to solve these problems, the present inventors have developed [ ¹¹ C] methylation on a carbon nucleus using ¹¹ CH ₃ I. Since all organic compounds have carbon atoms in their structures, the use of short-lived radioactive carbon ¹¹ C is an ideal radionuclide for making PET molecular probes of low-molecular organic compounds. Because it is considered.

Up to now, the present inventors have used not only high-speed C- [ ¹¹ C] methylation on an aromatic ring but also olefins by applying a Stille-type coupling reaction to an organotin compound as an intermediate raw material. Fast C- [ ¹¹ C] methylation on alkynes and heteroaromatic rings has also become possible (Patent Documents 1 and 2 and Non-Patent Documents 1 to 3). Furthermore, a new high-speed C- [ ¹¹ C] methylation reaction using organoboron compounds has been developed in addition to the conventional high-speed C- [ ¹¹ C] methylation of organotin compounds (patented) Literature 3, non-patent literature 4).

 On the other hand, many compounds having an isoquinoline ring are known in pharmaceuticals and physiologically active substances. Among them, compounds in which the 5-position of the isoquinoline ring is substituted with cyclic aminosulfonyl are circulatory organ agents (vasodilators, cerebral circulation improving agents, angina pectoris, cerebral cardiovascular thrombosis prophylactic and therapeutic agents. It is useful as a prophylactic and therapeutic agent for hypertension and a prophylactic and therapeutic agent for glaucoma) (Patent Documents 4 to 7) and has been found to have a protein kinase inhibitory action (Non-Patent Document 5).

 Hidaka et al. Of the present inventors invented fasudil [hexahydro-1- (5-isoquinolinesulfonyl) -1H-1,4-diazepine], the world's first and only Rho kinase inhibitor clinically applied, and arachnoid membrane Although it was provided to humans as a therapeutic agent for cerebral vasospasm after lower hemorrhage, fasudil is a compound in which the 5-position of the isoquinoline ring is substituted with cyclic aminosulfonyl (Patent Document 6). Furthermore, compounds in which the 5-position of the isoquinoline ring is substituted with a cyclic aminosulfonyl and the 4-position has a substituent such as a methyl group are also disclosed (Patent Documents 7 to 9).

 Among the compounds having the structural characteristics, H-1152P [(S)-(+)-hexahydro-2-methyl-1-[(4-methyl-5--5] discovered by Hidaka, the present inventor. Isoquinolinyl) sulfonyl] -1H-1,4-diazepine] is known as a selective and strong Rho kinase inhibitor (Patent Document 6, Non-Patent Document 6).

 Rho kinase is a serine / threonine protein kinase that has been identified as a target protein for Rho, a low molecular weight GTP-binding protein, and is used in various cell types such as cell migration, cell proliferation, cell contraction, cell adhesion, and cell motility. It is an important enzyme related to function.

 Rho kinase inhibitors such as fasudil and H-1152P are widely used in studies to investigate the role of Rho kinase in disease, and as a result, Rho kinase inhibitors are used in various diseases such as cerebral vasospasm, coronary artery spasm, Hypertension, pulmonary hypertension, asthma, glaucoma, renal disorder, angina pectoris, myocardial infarction, arteriosclerosis, restenosis, stroke, heart failure, myocardial hypertrophy, ischemia reperfusion disorder, vascular endothelial disorder, inflammatory bowel disease, prostate Hypertrophy, Raynaud's disease, neuropathic pain, Alzheimer's disease, Huntington's chorea, spinal cord injury, epilepsy, multiple sclerosis, osteoporosis, erectile dysfunction, liver disorders, cancer, wound healing, insulin resistance, diabetes, etc. It has been reported to be useful as a therapeutic agent. With regard to cancer, it has been reported that expression of Rho kinase is increased with rapid progression and strong invasive cancer and metastatic cancer, and it is known that Rho kinase is involved in metastasis and invasion. These cancers include sarcoma, Glioblastoma, glioma, head and neck cancer, hepatocellular carcinoma, clear cell renal cell carcinoma, lung cancer, ovarian cancer, breast cancer, esophageal cancer, colon cancer, bladder cancer , Prostate cancer, malignant melanoma.

 Compounds in which the 5-position of the isoquinoline ring is substituted with cyclic aminosulfonyl have been found not only to inhibit Rho kinase but also to compounds that inhibit various protein kinases such as PKA, PKG, AURKA, and CAMK2 (Non-patent Document 7). ), It can be used not only as a tool for protein kinase research but also as a medicine.

Therefore, if an [ ¹¹ C] methyl group can be introduced into the isoquinoline ring of an isoquinoline derivative having a specific physiological activity such as protein kinase inhibition by a fast C- [ ¹¹ C] methylation reaction, analysis by PET images becomes possible. Analysis of the brain dynamics and the potential as a drug (therapeutic drug). Furthermore, it can be used as a basic technology for biological research using these protein kinase inhibitors and clinical research using animal / human PET.

WO / 2007/046258 WO / 2010/074272 WO2008 / 023780 JP 57-156463 A JP 58-121279 A JP 61-227581 US2008 / 116481 WO1997 / 028130 WO2006 / 115247

Suzuki, M., et al., Chem. Eur. J., 3, 2039-2042 (1997). Hosoya, T., et al., Org. Biomol. Chem., 4, 410-415 (2006). Hosoya, T., et al., Org. Biomol. Chem., 2, 24-27 (2004). Doi, H., et al., Chem. Eur. J., 15,4165-4171 (2009). Hidaka, H et al., Biochemistry, 23, 5036-5041 (1984). Sasaki, Y., et al., Pharmacol. Ther. 93, 225-232 (2002). Tamura, M., et al., Biochim Biophys Acta, 1754,245-252 (2005).

However, since the half-life of ¹¹ C is only 20 minutes, the introduction of ¹¹ C requires a high speed, and it has been common technical knowledge so far to be carried out at the final stage of the synthesis. In addition, in order to secure the measurement time (about 1 hour), there is a restriction that the reaction, purification, and administration to the living body are performed within 40 minutes, which is twice the half-life. For this reason, the time allowed for the introduction of ¹¹ C is only about 5 minutes. Furthermore, the ¹¹ C nuclides that can be produced by the cyclotron are extremely small (several tens to several hundreds nmol level; taking into account the inclusion of ¹² C), and are very large for chemical reaction with ultra-diluted ¹¹ C nuclides. It is carried out under special conditions in the presence of an excess of the labeled substrate. For this reason, the most important issue is how efficiently bioactive organic compounds and drug discovery candidate compounds can be converted into PET molecular probes in a short time.

 The above-mentioned fast methylation method (Patent Documents 1 and 2 and Non-Patent Documents 1 to 3) developed by the present inventor Suzuki et al. Is a method that can be a powerful tool for solving these problems. It cannot be a general purpose method. This is because the functional group present in the substrate to be methylated may inhibit the reaction.

This is remarkable in the above-mentioned [ ¹¹ C] methylation of the compound in which the 5-position of the isoquinoline ring is substituted with cyclic aminosulfonyl. That is, when trying to [ ¹¹ C] methylate the isoquinoline skeleton by the fast methylation reaction described above, the chemically active secondary amino group present in the precursor inhibits the methylation reaction. In addition, the secondary amino group is [ ¹¹ C] methylated. Therefore, it is usually considered that the secondary amino group of the precursor is protected with a protecting group and deprotected after [ ¹¹ C] methylation. However, as described above, since a series of operations from introduction of ¹¹ C to administration to a living body requires high speed, it is common knowledge of those skilled in the art to perform [ ¹¹ C] methylation reaction at the final stage of synthesis. It was difficult to perform the deprotection reaction after [ ¹¹ C] methylation in terms of time and technology. For this reason, in the case of a compound in which the 5-position of the isoquinoline ring is substituted with a cyclic aminosulfonyl in spite of being a useful compound, the introduction of a [ ¹¹ C] methyl group onto the carbon of the isoquinoline ring has been conventionally performed. It wasn't.

The present invention has been made in view of the above-described conventional circumstances, and is for producing an ¹¹ C-labeled isoquinoline derivative in which an [ ¹¹ C] methyl group is introduced into an isoquinoline skeleton, a method for producing the ¹¹ C-labeled isoquinoline derivative, and the ¹¹ C-labeled isoquinoline derivative. It aims at providing the precursor which can be used conveniently.

In order to solve the above problems, the present inventors have conducted intensive research, and an ¹¹ C-labeled isoquinoline derivative represented by the following general formula (1) is represented by the following general formula (2) according to the production method described later. It was found that the precursor can be produced in a short time and with a high yield. Further, a molecular probe for PET was prepared using the compound of the general formula (1) and administered to a living body, and the tissue was imaged by PET. As a result, the compound of the general formula (1) was found to be useful as a radiopharmaceutical and a radiodiagnostic agent for PET, and the present invention was completed.
That is, the present invention provides the following inventions.

The present invention provides an ¹¹ C-labeled isoquinoline derivative represented by the following general formula (1) or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

(Where
One of R ¹ and R ² represents a [ ¹¹ C] methyl group, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group, a nitro group, a fluoro group, or Represents an aryl group;
R ³ and R ⁴ each independently represent a hydrogen atom, alkyl group, alkenyl group, amino group, alkylamino group, dialkylamino group, aminoalkyl group, halogenoalkyl group, alkanoyl group, aminoalkanoyl group or alkylaminoalkanoyl group. Or R ³ and R ⁴ together form an alkylene group or an alkenylene group and are bridged between any two carbons in the cyclic diamino group in general formula (1) Yes;
R ⁵ represents a hydrogen atom, an aminoalkanoyl group or an alkylaminoalkanoyl group;
m represents a natural number of 1 to 4, and n represents a number of 2 or 3. )

 The present invention also provides a suitable precursor for producing the compound of the general formula (1), which is represented by the following general formula (2).

(Where
R ⁶ and R ⁷ either represents a trialkyltin group, a boronyl group or a dialkoxyboryl group, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group, Represents a nitro group, a fluoro group or an aryl group;
R ³ and R ⁴ are the same as above;
R ⁸ represents tert-butoxycarbonyl group, trihaloacetyl group, tert-butoxycarbonylaminoalkanoyl group, trihaloacetylaminoalkanoyl group, N-alkyl-N-tert-butoxycarbonylaminoalkanoyl group or N-alkyl-N-trihaloacetylamino. Represents an alkanoyl group;
m represents a natural number of 1 to 4, and n represents a number of 2 or 3. )

In addition, the present invention introduces a [ ¹¹ C] methyl group on the isoquinoline skeleton in the precursor (2) of the general formula (1) and then removes a protective group such as a trihaloacetyl group under basic conditions. Provided is a method for easily producing a ¹¹ C-labeled isoquinoline derivative represented by the general formula (1), which is characterized by a high yield in a short time.

The present invention also provides a PET probe containing an ¹¹ C-labeled isoquinoline derivative represented by the general formula (1).

Furthermore, the present invention provides a tissue imaging method characterized by administering an ¹¹ C-labeled isoquinoline derivative of the general formula (1) to a living body and examining the distribution of ¹¹ C in the living body.

3 is a preparative HPLC chart of a reaction filtrate in Example 3.

In general formula (1), ^one of R ¹ and R ² represents a [ ¹¹ C] methyl group, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group. , A nitro group, a fluoro group or an aryl group.

Here, the alkyl group is a linear, branched, or cyclic alkyl group having 1 to 8 carbon atoms (C _1-8 alkyl group) (here, “C _1-8 ” means 1 carbon atom). And the like, the same applies hereinafter), an alkyl group having 1 to 6 carbon atoms is preferable, and an alkyl group having 1 to 3 carbon atoms is more preferable.
Specific examples include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, isopentyl group, n-hexyl group, Examples thereof include an isohexyl group and a cyclopropyl group. Of these, those having 1 to 3 carbon atoms are preferable, and methyl group and ethyl group are particularly preferable.

The halogenoalkyl group is preferably a halogeno C _1-8 alkyl group, more preferably a halogeno C _1-6 alkyl group. Specific examples include chloromethyl group, fluoromethyl group, chloroethyl group, fluoroethyl group, trifluoromethyl group and the like.

Examples of the alkenyl group include a linear or branched alkenyl group having 2 to 8 carbon atoms (C _2-8 alkenyl group), and an alkenyl group having 2 to 6 carbon atoms is preferable. Specific examples include vinyl group, allyl group, isopropenyl group, 2-methallyl group, 2-butenyl group, and 3-butenyl group. Of these, those having 2 to 4 carbon atoms are preferred.

Examples of the alkoxy group include a linear or branched alkoxy group having 1 to 8 carbon atoms (C _1-8 alkoxy group), and an alkoxy group having 1 to 6 carbon atoms is preferable. Specific examples include methoxy group, ethoxy group, n-propoxy group, isopropoxy group, n-butoxy group, isobutoxy group, sec-butoxy group, and tert-butoxy group.

As an alkylthio group, a C1-C8 linear or branched alkylthio group ( _C1-8 alkylthio group) is mentioned, A C1-C6 alkylthio group is preferable. Specific examples include a methylthio group, an ethylthio group, an isopropylthio group, and an n-propylthio group.

Examples of the aryl group include a C _6-14 aryl group, and a phenyl group and a naphthyl group are preferable, and a phenyl group is more preferable.

As R ¹ and R ² , either one represents a [ ¹¹ C] methyl group, and the other is a hydrogen atom, a C _1-8 alkyl group, a nitro group, a fluoro group, a cyano group, a halogeno C _1-8 alkyl group, It is preferably a phenyl group or a C _2-8 alkenyl group. The other of the [ ¹¹ C] methyl group is more preferably a hydrogen atom, a cyano group, a C _1-6 alkyl group or a halogeno C _1-6 alkyl group. Further, the other of the [ ¹¹ C] methyl group is preferably a hydrogen atom or a C _1-3 alkyl group.

R ¹ may be substituted at any of the 1-position, 3-position and 4-position of the isoquinoline skeleton. R ² may be substituted at any of the 6-position, 7-position and 8-position of the isoquinoline skeleton. However, the substitution position of the [ ¹¹ C] methyl group is particularly preferably the 4-position of the isoquinoline ring.

R ³ and R ⁴ are each independently a hydrogen atom, alkyl group, alkenyl group, amino group, alkylamino group, dialkylamino group, aminoalkyl group, halogenoalkyl group, alkanoyl group, aminoalkanoyl group or alkylaminoalkanoyl group. R ³ and R ⁴ may form an alkylene group or an alkenylene group, and may be bridged between any two carbons in the cyclic diamino group in the general formula (1).

Examples of the alkyl group, alkenyl group, and halogenoalkyl group include those shown as examples of R ¹ and R ² .
As the alkylamino group, a C _1-8 alkylamino group is preferable, and specific examples include methylamino group, ethylamino group, n-propylamino group, isopropylamino group, n-butylamino group, isobutylamino group, sec- A butylamino group, an n-pentylamino group, an n-hexylamino group and the like can be mentioned. The dialkylamino group is preferably a di-C _1-8 alkylamino group, and specific examples include a dimethylamino group, a diethylamino group, a dipropylamino group, a dibutylamino group, and the like. The aminoalkyl group is preferably an amino C _1-8 alkyl group, and specific examples thereof include aminomethyl group, aminoethyl group, aminopropyl group, aminobutyl group and the like.

Examples of the alkanoyl group include a linear or branched alkanoyl group having 2 to 8 carbon atoms (C _2-8 alkanoyl group), and an alkanoyl group having 2 to 6 carbon atoms is preferable. Specific examples include an acetyl group, a propionyl group, and a butyryl group. Examples of the aminoalkanoyl group include an amino-C _2-8 alkanoyl group, and an amino-C _2-6 alkanoyl group is preferable. Specific examples include aminoacetyl group, aminopropionyl group, aminobutyryl group and the like.

Examples of the alkylaminoalkanoyl group include a C _1-8 alkylamino C _2-8 alkanoyl group, and a C _1-4 alkylamino C _2-4 alkanoyl group is preferable. Specific examples thereof include a methylaminoacetyl group and a methylaminopropionyl group. Groups and the like. Examples of the alkoxycarbonyl group include a C _1-8 alkoxycarbonyl group, and examples thereof include a methoxycarbonyl group and an ethoxycarbonyl group.

Examples of the alkylene group formed by combining R ³ and R ⁴ include C _1-3 alkylene groups such as a methylene group, an ethylene group, and a trimethylene group (—CH ₂ CH ₂ CH ₂ —). Group and ethylene group are preferred. Examples of the alkenylene group formed by combining R ³ and R ⁴ include C _2-4 alkenylene groups such as —CH═CH— and —CH ₂ CH═CH—. These alkylene groups or alkenylene groups may be bridged between two carbons to any position on the nitrogen-containing saturated heterocyclic ring in formula (1). Such crosslinking, crosslinked C _1-3 alkylene group, i.e. _{-CH 2 -, - CH 2 CH} 2 -, - CH 2 CH 2 CH 2 - by crosslinking is preferable.

R ³ and R ⁴ are each independently a hydrogen atom, a C _1-8 alkyl group, a C _2-8 alkenyl group, an amino group, an amino C _1-8 alkyl group, a halogeno C _1-8 alkyl group, C _{1 -8} alkylamino group, di-C _1-8 alkylamino group, C _2-8 alkanoyl group or amino C _2-8 alkanoyl group, or R ³ and R ⁴ together form a bridged C _1- Preferably ₃ alkylene groups are formed.
R ³ and R ⁴ are each independently a hydrogen atom, a C _1-8 alkyl group, an amino group, a C _1-8 alkylamino group, an amino C _1-8 alkyl group or a halogeno C _1-8 alkyl group. More preferably, R ³ and R ⁴ together form a bridged C _1-3 alkylene group.
Further, R ³ and R ⁴ are preferably a hydrogen atom, a C _1-6 alkyl group or a halogeno C _1-6 alkyl group.

R ⁵ represents a hydrogen atom, an aminoalkanoyl group or an alkylaminoalkanoyl group. The aminoalkanoyl group or alkylaminoalkanoyl group is the same as described above.

 m represents a natural number of 1 to 4, n represents 2 or 3, m is preferably a natural number of 1 to 3, and n is particularly preferably 2.

The ¹¹ C-labeled isoquinoline derivative of the present invention includes salts, hydrates, solvates and prodrugs thereof in addition to the compound represented by the above general formula (1). Here, the prodrug refers to a compound that is hydrolyzed in vivo to produce a compound represented by the general formula (1). Such prodrugs include compounds produced by all prodrugation techniques known to those skilled in the art. For example, a compound in which an amino group is derivatized to an amide group that can be easily hydrolyzed in vivo is exemplified. Specific examples include compounds in which the amino group present in the general formula (1) is derived from acetamide or the like. Since these prodrugs can be easily produced from the ¹¹ C-labeled isoquinoline derivative of the above general formula (1) in a short time, there is no significant obstacle to application as a probe for PET.

 Examples of the pharmaceutically acceptable salt include acid addition salts for amino groups present in the general formula (1). Examples of acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulfate, and nitrate, formate, acetate, propionate, maleate, fumarate, and succinic acid. Organic salts such as salt, lactate, malate, tartrate, citrate, ascorbate, malonate, oxalate, glycolate, phthalate, benzenesulfonate, etc. A combination of the above-mentioned salts can also be used.

Next, as the compound represented by the general formula (2), in the formula (2), R ^3-4 , m and n are the R shown in the ¹¹ C-labeled isoquinoline derivative represented by the general formula (1). ^3-4 Same as m, n.

R ⁸ includes tert-butoxycarbonyl group, trihaloacetyl group, tert-butoxycarbonylaminoalkanoyl group, trihaloacetylaminoalkanoyl group, N-alkyl-N-tert-butoxycarbonylaminoalkanoyl group or N-alkyl-N-trihalo. An acetylaminoalkanoyl group is shown.

 As the trihaloacetyl group, a trifluoroacetyl group, a trichloroacetyl group, or a tribromoacetyl group can be used, but a trifluoroacetyl group that can be easily detached is preferable.

The trihaloacetylaminoalkanoyl group and the N-alkyl-N-trihaloacetylaminoalkanoyl group are obtained by protecting the amino group of the above aminoalkanoyl group or alkylaminoalkanoyl group with a trihaloacetyl group, and as a trihaloacetylaminoalkanoyl group Includes a trihaloacetylamino-C _2-8 alkanoyl group, and a trihaloacetylamino-C _2-6 alkanoyl group is preferable. Specific examples include trifluoroacetylaminoacetyl group, trifluoroacetylaminopropionyl group, trifluoroacetylaminobutyryl group and the like. Examples of the N-alkyl-N-trihaloacetylaminoalkanoyl group include N—C _1-8 alkyl-N-trihaloacetylamino C _2-8 alkanoyl group, and N—C _1-4 alkyl-N-trihaloacetylamino. A C _2-4 alkanoyl group is preferred, and specific examples include N-methyl-N-trifluoroacetylaminoacetyl group, N-methyl-N-trifluoroacetylaminopropionyl group and the like.
Similarly, as the tert-butoxycarbonylaminoalkanoyl group and the N-alkyl-N-tert-butoxycarbonylaminoalkanoyl group, the amino group of the aminoalkanoyl group or the alkylaminoalkanoyl group is protected with a tert-butoxycarbonyl group. Indicates.

R ⁶ and R ⁷ either represents a trialkyltin group, a boronyl group or a dialkoxyboryl group, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group, A nitro group, a fluoro group or an aryl group is shown.

R ⁶ may be substituted at any of the 1-position, 3-position and 4-position of the isoquinoline skeleton.
R ⁷ may be substituted at any of the 6-position, 7-position and 8-position of the isoquinoline skeleton. However, the substitution position of the trialkyltin group, boronyl group or dialkoxyboryl group is particularly preferably the 4-position of the isoquinoline ring.

As R ⁶ and R ⁷ , either one represents a trialkyltin group, a boronyl group or a dialkoxyboryl group, and the other represents a hydrogen atom, a C _1-8 alkyl group, a nitro group, a fluoro group, a cyano group, a halogeno C _It is preferably a _1-8 alkyl group, a phenyl group or a C _2-8 alkenyl group. More preferably, the other of the trialkyltin group, boronyl group or dialkoxyboryl group is a hydrogen atom, a cyano group, a C _1-6 alkyl group or a halogeno C _1-6 alkyl group. Further, the other of the trialkyltin group, boronyl group or dialkoxyboryl group is preferably a hydrogen atom or a C _1-3 alkyl group.

Examples of the trialkyl tin group include a tri C _1-6 alkyl tin group, and a tri C _2-4 alkyl tin group is preferable, and specific examples include a triethyl tin group, a tripropyl tin group, or a tributyl tin group.

The dialkoxy boryl group, include di-C _1-6 Arukokishiboriru group, di C _1-3 Arukokishiboriru group is preferable, specific examples dimethoxy boryl group include Jietokishiboriru group or dipropyl alkoxy boryl group. Further, two alkoxy groups may be combined to form alkylene or phenylene. Specific examples include 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl group, 4 4,6,6-tetramethyl-1,3,2-dioxaborin-2-yl group or benzo [d] [1,3,2] dioxaborol-2-yl group.

<Production method>
The ¹¹ C-labeled isoquinoline derivative represented by the general formula (1) is synthesized from, for example, Scheme 1 via the compound represented by the general formula (2) from the compound (3) produced by the method described later. It can be manufactured according to the method shown.

(In the formula, one of R ⁹ and R ¹⁰ represents a halogen atom, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group, a nitro group, a fluoro group, or An aryl group is shown.
R ^1-8 , m and n are the same as above. )

[First step]
Compound (4) is prepared by using a tert-butoxycarbonylating agent such as di-tert-butyldicarbonate or a trihaloacetyl such as halogenated trihaloacetyl or trihaloacetic anhydride in the presence of a base in a solvent suitable for compound (3). It can be produced by reacting an agent.

 The reaction solvent is not particularly limited as long as it does not interfere with the reaction. For example, ethers such as tetrahydrofuran, dioxane and diethyl ether, hydrocarbons such as benzene and toluene, halogenated hydrocarbons such as methylene chloride and chloroform, Aprotic solvents such as N, N-dimethylformamide and N, N-dimethylacetamide, pyridine, acetonitrile, acetone, water, or a mixture thereof can be used.

 This reaction is preferably carried out in the presence of a suitable base. Such bases include alkali metals such as alkali metal bicarbonates (eg, sodium bicarbonate), alkali metal carbonates (eg, potassium carbonate), alkali metal hydroxides (eg, sodium hydroxide, potassium hydroxide), Organic tertiary amines such as triethylamine and triethylenediamine can be used. If a basic solvent such as pyridine is used as the solvent, such a base is unnecessary and preferable.

 Usually, this reaction often proceeds at room temperature, but it can be carried out at −78 to 150 ° C., preferably 0 to 120 ° C. by cooling or heating as necessary. The reaction time varies depending on the raw materials used, solvent, reaction temperature and the like, but is usually 5 minutes to 70 hours.

[Second step]
Compound (2) can be produced by converting the halogen atom of compound (4) into a trialkyltin group, boronyl group or dialkoxyboryl group by a method known per se.
That is, the halogen compound represented by the general formula (4) is converted into a Grignard reagent using magnesium or a lithiation using an organometallic reagent such as alkyllithium in an ether solvent such as tetrahydrofuran, dioxane, or diethyl ether. The trialkyltin body or boronyl body represented by the compound (2) can be produced by activating and subsequently treating with a trialkyltin chloride or an organoboron compound corresponding to R ⁶ and R ⁷ . In addition, the halogen compound represented by the general formula (4) is converted into a trimethyl compound represented by the general formula (2) by a stannation or boronation reaction using a bis (trialkyltin) compound or a diborane compound in the presence of a palladium catalyst. Alkyl tin bodies, boronyl bodies, and dialkoxyboryl bodies can be produced. This reaction can be carried out according to a known method.

As the halogen represented by R ⁹ and R ¹⁰ , chlorine, bromine, or iodine is preferable.

[Third step]
A trialkyltin compound, boronyl compound or dialkoxyboryl compound represented by compound (2) is converted into [11C] methyl iodide in an aprotic polar solvent in the presence of a ligand, a palladium complex and a base. By reacting, the [11C] methyl compound represented by the compound (5) can be produced.
[11C] As the method of methylation, the method of Patent Document 1, 2, or 3 described above can be used.

Examples of the reaction solvent include hydrocarbons such as benzene and toluene, halogenated hydrocarbons such as methylene chloride and chloroform, N, N-dimethylformamide, N, N-dimethylacetamide, and N—C _1-6 alkyl-2- An aprotic polar solvent such as pyrrolidinone, pyridine, acetonitrile, water, or a mixture thereof can be used. Of these, aprotic polar solvents such as N, N-dimethylformamide and N—C _1-6 alkyl-2-pyrrolidinone are preferred. Particularly preferred is N—C _1-3 alkyl-2-pyrrolidinone, specifically N-methyl-2-pyrrolidinone (NMP), N-ethyl-2-pyrrolidinone or N-propyl-2-pyrrolidinone. It is done.

 As the ligand, a phosphine ligand such as trialkylphosphine or triarylphosphine or an arsine ligand such as triarylarsine can be used. Usually, in this reaction, since the bulk of the ligand forms a highly active reaction field, phosphine ligands such as tri-o-tolylphosphine and (di-tert-butyl) methylphosphine are particularly preferable.

 As the palladium complex, generally known zero-valent palladium complexes such as tetrakistriphenylphosphine palladium and divalent palladium complexes such as palladium chloride and palladium acetate can be used. Of these, a zero-valent palladium complex is preferable, but tris (dibenzylideneacetone) dipalladium is particularly preferable.

 Bases include alkali metal bicarbonate (eg, sodium bicarbonate), alkali metal carbonate (eg, potassium carbonate, cesium carbonate), alkali metal phosphate (eg, potassium phosphate), alkali metal hydroxide ( Examples thereof include inorganic bases such as sodium hydroxide and potassium hydroxide, and organic tertiary amines such as triethylamine and ethyldiisopropylamine.

[Fourth step]
^{The 11} C-labeled isoquinoline derivative (1) can be produced by removing the tert-butoxycarbonyl group or trihaloacetyl group of the compound (5) by acid treatment or alkali treatment. The reaction may follow a method known per se. For example, a tert-butoxycarbonyl group can be removed by reacting an acid such as hydrochloric acid or trifluoroacetic acid in a suitable reaction solvent. In the case of a trihaloacetyl group, it can be removed by a hydrolysis reaction or a solvolysis reaction using an inorganic base such as an alkali metal hydrogen carbonate, an alkali metal carbonate, or an alkali metal hydroxide.

 Compound (3) shown in Scheme 1 can be synthesized by the methods described in Patent Documents 7 to 9 or similar methods. Moreover, it is also compoundable by the method shown to the following schemes 2-6.

(In the formula, L ¹ represents a hydroxyl group or a leaving group, R ¹¹ represents an N protecting group, and R ³ , R ⁴ , R ⁹ , R ¹⁰ , m and n are the same as described above.)

Examples of L ¹ as a leaving group include residues of reactive derivatives of sulfonic acid described later. Examples of the protecting group used for N include acyl groups such as formyl, acetyl and benzoyl, aralkyloxycarbonyl groups such as benzyloxycarbonyl, alkoxycarbonyl groups such as tert-butyloxycarbonyl, and aralkyl groups such as benzyl. Can do.

 The amine represented by the general formula (7) is reacted with the sulfonic acid represented by the general formula (6) or a reactive derivative thereof in a suitable solvent, and the protective group is removed as necessary to obtain the compound (3a) Can be manufactured.

 As the reactive derivative of sulfonic acid, sulfonic acid halide (eg, sulfonic acid fluoride, sulfonic acid chloride, sulfonic acid bromide), sulfonic acid anhydride, N-sulfonylimidazolide and the like are used. In particular, a sulfonic acid halide is preferable.

(In the formula, L ² represents a leaving group, and R ³ , R ⁴ , R ⁹ , R ¹⁰ , m and n are the same as described above.)

Examples of L ² as a leaving group include halogens such as chlorine or bromine, acyloxy groups such as acetyloxy, or sulfonyloxy groups such as mesyloxy and tosyloxy.
Compound (3a) can be produced by reacting compound (8) with amine, guanidine or ammonia. This reaction can be performed according to a known method (Acta Chemica Scand., 45, 621 (1991)).

(Wherein R ¹² represents a hydrogen atom or an amino-protecting group, and L ² , R ³ , R ⁴ , R ⁹ , R ¹⁰ , m and n are the same as described above.)

 Compound (3a) can be produced by reacting compound (9) with compound (10) and, if necessary, treating with acid or alkali to remove the protecting group. This reaction can be performed according to a known method (Acta Chemica Scand., 45, 621 (1991)).

(Wherein R ¹² represents a hydrogen atom or an amino-protecting group, and L ² , R ³ , R ⁴ , R ⁹ , R ¹⁰ , m and n are the same as described above.)
Compound (3a) is obtained by subjecting compound (11) to an intramolecular dehydration reaction (so-called Mitsunobu reaction) using an organic phosphorus reagent such as triphenylphosphine and an azo reagent such as diethyl azodicarboxylate or diisopropyl azodicarboxylate. It can be prepared by removing the protecting group R ¹² .

(In the formula, L ³ represents a hydroxyl group or a leaving group, R ¹³ represents a hydrogen atom or a C _1-7 alkyl group, and l represents a natural number of 1 to 7. Also, R ³ , R ⁴ , R ⁹ , R ¹⁰ , R ¹² , m and n are the same as above.)

 The amine represented by the general formula (3a) is reacted with the carboxylic acid represented by the general formula (12) in the presence of a suitable condensing agent, and the protective group is removed as necessary to produce the compound (3b). be able to. Alternatively, general formula (3a) is reacted with a reactive derivative of carboxylic acid in the presence of a suitable base to produce compound (3b).

 Examples of the condensing agent include carbodiimide condensing agents such as dicyclohexylcarbodiimide and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, 1H-benzotriazol-1-yloxytripyrrolidinophosphonium and hexafluorophosphate. A phosphonium condensing agent or the like is used.

Examples of L ³ as a leaving group include residues of reactive derivatives of carboxylic acids. As the reactive derivative of carboxylic acid, carboxylic acid halide (eg, carboxylic acid chloride, carboxylic acid bromide), carboxylic acid anhydride, carboxylic acid azide, and active ester are used.

^{Some 11} C-labeled isoquinoline derivatives (1) have an asymmetric carbon and have optical isomers. Any of these isomers and mixtures thereof are included in the present invention. The optical isomer can be obtained by using an optically active raw material compound (S configuration or R configuration). The racemate has pharmacological activity as it is, but can be resolved into each isomer as desired. For example, a mixture of isomers can be prepared by a known optical resolution method such as optically active carboxylic acid (eg, (+)-or (-)-tartaric acid, (+)-or (-)-malic acid) or optically active sulfone. A salt with an acid (eg, (+)-camphorsulfonic acid) can be produced and separated by a method of fractional crystallization or a method using an optically active column.
^{The 11} C-labeled isoquinoline derivative (1) can form a salt by a known method. For example, the hydrochloride can be obtained by dissolving the compound of the present invention in an alcohol solution or ethyl ether solution of hydrogen chloride.

The ¹¹ C-labeled isoquinoline derivative (1) which is the compound of the present invention can be suitably used as a molecular probe for PET, as will be described later. In this case, the administration route can be oral or parenteral. Examples of the dosage form include tablets, capsules, granules, powders, injections, eye drops and the like, and these can be used in combination with commonly used techniques. However, in view of the half life of ¹¹ C, injections and eye drops that do not require time for preparation are preferred.
In the case of an injection or eye drop, an aqueous medium such as sterile purified water, physiological saline or isotonic solution, or a non-aqueous medium such as sesame oil, soybean oil or olive oil is used as the liquid medium. It may be an emulsion containing both an aqueous medium and an oily medium.
Parenteral preparations such as injections and eye drops include, for example, isotonic agents such as glycerin, propylene glycol, sodium chloride, potassium chloride, sorbitol, mannitol, phosphoric acid, phosphate, citric acid, glacial acetic acid, ε- Buffers such as aminocaproic acid and trometamol, pH regulators such as hydrochloric acid, citric acid, phosphoric acid, glacial acetic acid, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, polysorbate 80, polyoxyethylene hydrogenated castor oil 60 , Solubilizing aids such as macrogol, purified egg yolk lecithin, purified soybean lecithin, emulsifiers such as polyoxyethylene (160) polyoxypropylene (30) glycol, cellulose polymers such as hydroxypropyl methylcellulose, hydroxypropylcellulose, polyvinyl alcohol , Polyvinyl pylori Thickeners such as sodium chloride, stabilizers such as edetic acid and sodium edetate, general-purpose sorbic acid, potassium sorbate, benzalkonium chloride, benzethonium chloride, methyl paraoxybenzoate, propyl paraoxybenzoate, chlorobutanol, etc. A preservative or preservative, a soothing agent such as chlorobutanol, benzyl alcohol, lidocaine and the like can be prepared by combining with the compound of the present invention as necessary.

 In the case of injections or eye drops, the pH is preferably set to 4.0 to 8.0, and the osmotic pressure ratio is preferably set to around 1.0.

 By using this PET probe as a radiopharmaceutical or diagnostic composition, it becomes possible to analyze protein kinase inhibitors using PET images, and to analyze the dynamics in the brain and the potential as a drug (therapeutic) Is possible. Furthermore, it can be used as a basic technology for biological research and clinical research using animal / human PET.

That is, tissue can be imaged by administering the ¹¹ C-labeled isoquinoline derivative of the present invention to a living body and examining the distribution of ¹¹ C in the living body. ^{For the 11} C distribution, a PET method for detecting positrons can be used. If, for example, the brain is selected as the target for imaging such tissues, it will be possible to capture ¹¹ C-labeled isoquinoline derivatives bound to protein kinases in the brain, and in turn, gain a lot of knowledge about the functions of protein kinases in the brain. be able to.

For example, since H-1152P is a specific inhibitor of Rho kinase, the compound of the present invention, which is its ¹¹ C form, is administered to experimental rats, etc. by intravenous injection, etc. By photographing, it becomes possible to capture the distribution of Rho kinase in the brain and the ¹¹ C-labeled isoquinoline derivative bound to the Rho kinase in the brain, and gain a lot of knowledge about the role of Rho kinase in the brain, etc. Can do.

Furthermore, PET images taken after administration of a PET formulation containing the ¹¹ C-labeled isoquinoline derivative of the present invention to humans should be used for diagnosis of diseases associated with abnormal protein kinase distribution and diseases associated with protein kinases. Can do. Such protein kinases include, but are not limited to, Rho kinase, PKA, PKG, AURKA, CAMK2.

For example, since H-1152P is a specific inhibitor of Rho kinase, PET images taken after administration of the compound of the present invention, which is its ¹¹ C form, to humans show diseases associated with abnormal distribution of Rho kinase and Rho It can be used for diagnosis of diseases related to kinases.
Rho kinase related diseases include cerebral vasospasm, coronary artery spasm, hypertension, pulmonary hypertension, asthma, glaucoma, kidney damage, angina, myocardial infarction, arteriosclerosis, restenosis, stroke, heart failure, myocardial hypertrophy, Ischemia reperfusion disorder, vascular endothelial disorder, inflammatory bowel disease, prostatic hypertrophy, Raynaud's disease, neuropathic pain, Alzheimer's disease, Huntington's chorea, spinal cord injury, epilepsy, multiple sclerosis, osteoporosis, erectile dysfunction, Includes liver damage, cancer, wound healing, insulin resistance, and diabetes. The above cancers include sarcoma, Glioblastoma, glioma, head and neck cancer, hepatocellular carcinoma, clear cell renal cell carcinoma, lung cancer, ovarian cancer, breast cancer, esophageal cancer, colon cancer, bladder cancer , Prostate cancer, and malignant melanoma.

In addition, since detection of ¹¹ C-labeled isoquinoline derivative in the body with PET is extremely sensitive, it can also be applied to microdose clinical trials. This clinical trial is a method for administering pharmacokinetics etc. using high-sensitivity microanalysis by administering an extremely small amount (microdose) of a drug candidate substance to the human body. There is little influence on the human body. For this reason, the safety of pharmacological studies is extremely high, and in the microdose clinical study, the drug pharmacokinetics in the human body can be known by applying the drug candidate substances to humans safely and quickly before entering Phase I. Screening of candidate development substances from the aspect can be performed, and the non-clinical trial period can be greatly shortened.

 The present invention will be specifically described below with reference to examples. These illustrations are for better understanding of the present invention and do not limit the scope of the present invention.

Reference example 1
Synthesis of 4-trifluoroacetyl-{(S) -hexahydro-2-methyl-1- (4-bromo-5-isoquinolinesulfonyl)}-1H-1,4-diazepine (b)

(S) -Hexahydro-2-methyl-1- (4-bromo-5-isoquinolinesulfonyl) -1H-1,4-diazepine (a) synthesized according to the method of Patent Document 8 was protected with a trifluoroacetyl group.
Specifically, triethylamine (134 μL, 97.3 mg, 962 μmol) was added to (a) (246.6 mg, 641.4 μmol) in dry dichloromethane (2 mL), cooled to 0 ° C., and then trifluoroacetic anhydride (134 μL, 202 mg, 962 μmol) was added. After this mixed solution was stirred at 0 ° C. for 2 hours, the reaction mixture was added to distilled water (about 10 mL), and extracted with dichloromethane (3 × 15 mL). The organic layer was washed successively with distilled water (15 mL) and saturated brine (15 mL), and dried over anhydrous sodium sulfate. The desiccant was filtered off and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (silica gel 5 g, eluent: hexane: acetone (2: 1)) to give pale yellow oily compound (b) (266 mg, 554 μmol, 86.4%). Obtained.
1H NMR (500 MHz, CDCl ₃ , 25 ° C): δ = 9.21 (s, 1H), 8.99 (s, 1H), 8.50 (dd, 4J (H, H) = 1.0 Hz, 3J (H, H) = 7.5 Hz, 1H), 8.21 (dd, 4J (H, H) = 1.0 Hz, 3J (H, H) = 8.0 Hz, 1H), 7.67-7.71 (m, 1H), 3.90-4.36 (m, 4H) , 3.33-3.66 (m, 3H), 2.18-2.32 (m, 1H), 1.87-2.04 (m, 1H), 1.06 ppm (d, 3J (H, H) = 7.0 Hz, 3H); HRMS (EI) m / z: [M + H] ⁺ calcd for C ₉ H ₇ BrN 207.9762 found 207.9745; Anal. Calcd for C ₁₇ H ₁₇ BrF ₃ N ₃ O ₃ S: C, 72.51; H, 3.57; N, 8.75. Found : C, 72.74; H, 3.79; N, 8.70.

Example 1
<Synthesis of organotin precursor>
Synthesis of 4-trifluoroacetyl-[(S) -hexahydro-2-methyl-1- {4- (tri-n-butylstannyl) -5-isoquinolinesulfonyl}]-1H-1,4-diazepine (c)

Organotin precursor (c) was synthesized from compound (b).
That is, (b) (99.4 mg, 0.205 mmol) and dry THF (2.5 mL) were added to a dry 20 mL Schlenk type reaction tube under argon atmosphere. After cooling this mixed solution to −100 ° C., tert-butyllithium (279 μL, 1.47 M hexane solution, 0.410 mmol) was added dropwise over 20 minutes, and the mixture was stirred at the same temperature for 2 hours. To this reaction solution, a THF solution (1.0 mL) of hexamethylphosphoric triamide (HMPA) (214 μL, 220 mg, 1.23 mmol) and (nC ₄ H ₉ ) ₃ SnCl (317 μL, 381 mg, 1.17 mmol) Of THF (1.0 mL) was added in order, the temperature was gradually raised to room temperature, and the mixture was stirred for 10 hours. The resulting reaction solution was added to a cooled saturated aqueous ammonium chloride solution (about 2 mL), and extracted with ethyl acetate (3 × 10 mL). The organic layer was washed with saturated brine (20 mL) and then dried over anhydrous sodium sulfate. The desiccant was filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (silica gel 10 g, developing solvent; hexane: acetone (2: 1)) to obtain an orange oily compound (c) (96.1 mg, 0.138 mmol, 67.5%).
1H NMR (400 MHz, CDCl ₃ , 25 ° C): δ = 9.15 (s, 1H), 8.74 (s, 1H), 8.25 (dd, 4J (H, H) = 1.4, 3J (H, H) = 7.5 Hz, 1H), 8.12 (dd, 4J (H, H) = 1.2, 3J (H, H) = 8.0 Hz, 1H), 7.61 (m, 1H), 4.16-4.21 (m, 1H), 2.91-3.87 (m, 6H), 1.52-1.71 (m, 2H), 1.44-1.50 (m, 6H), 1.24-1.34 (m, 6H), 1.10-1.15 (m, 6H), 0.84 (d, 3J (H, H) = 5.3 Hz, 3H), 0.83 ppm (t, 3J (H, H) = 7.4 Hz, 9H); HRMS (EI ⁺ , 100% acetone): m / z: calcd for C ₂₅ H ₃₅ F ₃ N ₃ O ₃ SSn ([MC ₄ H ₉ ] ⁺ ) 634.1376; found, 634.1359.

Example 2
<Synthesis of organoboronic acid ester precursor>
(S) -2,2,2-trifluoro-1- [3-methyl-4- {4- (4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl) isoquinoline Synthesis of -5-ylsulfonyl} -1,4-diazepan-1-yl] ethane According to the method described in Patent Document 3, (S) -2,2,2-trifluoro-1- [3 -Methyl-4- {4- (4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl) isoquinolin-5-ylsulfonyl} -1,4-diazepan-1-yl] Ethane can be synthesized.

Example 3
<Synthesis 1 of ¹¹ C-labeled isoquinoline derivative (d) 1>
Synthesis of (S) -hexahydro-2-methyl-1-[(4- [ ¹¹ C] methyl-5-isoquinolinyl) sulfonyl] -1H-1,4-diazepine (d)

Based on the following synthesis method, the compound (c) of Example 1 was subjected to [ ¹¹ C] methylation at the 4-position of the isoquinoline skeleton, and then the trifluoroacetyl group was eliminated under basic conditions to thereby remove the isoquinoline skeleton. An ¹¹ C-labeled isoquinoline derivative (d) in which the methyl group at the 4-position was [ ¹¹ C] methylated was obtained. Details are shown below.

^{The 11} C nucleus was produced by a nuclear reaction of ¹⁴ N (p, α) ¹¹ C using a cyclotron CYPRIS HM-12S manufactured by Sumitomo Heavy Industries, Ltd. [ ¹¹ C] methyl iodide was synthesized by converting ¹¹ CO ₂ → ¹¹ CH ₃ OH → ¹¹ CH ₃ I in the order of ¹¹ CO ₂ gas using ¹¹ CO ₂ gas as a starting material, using a dedicated labeling synthesizer.

On the other hand, tris (dibenzylideneacetone) dipalladium (Pd ₂ (dba) ₃ ) (1.8 mg, 1.9 μmol) and tri-o-tolylphosphine (P (o-tolyl) ₃ ) (9.4 mg, 30.9 μmol) NMP (N-methylpyrrolidinone) solution (0.27 mL) was placed in the reaction vessel (A) of the labeling synthesizer and placed at room temperature. The solution in the reaction vessel (A) was placed 10-20 minutes before blowing [ ¹¹ C] methyl iodide ( ¹¹ CH ₃ I). In addition, an NMP solution (80 μL) of organotin precursor (c) (4.0 mg, 5.8 μmol), CuBr (0.5 mg, 3.9 μmol), and CsF (1.5 mg, 9.7 μmol) was prepared in the reaction vessel (B). And set at room temperature. Subsequently, [ ¹¹ C] methyl iodide was blown into the reaction vessel (A) at a gas flow rate of 60-80 mL / min, and then allowed to stand for 1 minute. The obtained solution was transferred to the reaction vessel (B), and the inside of the reaction vessel (A) was washed with a small amount of NMP solution (60 μL), and this washing solution was also transferred to the reaction vessel (B). Subsequently, the mixed solution in the reaction vessel (B) was heated at 80 ° C. for 5 minutes. Subsequently, 2M NaOH (400 μL) was added and heated at 100 ° C. for 1 minute. The obtained reaction solution was diluted with methanol (500 μL) and sodium phosphate buffer (10 mM, pH 7.4, 500 μL), and then filtered using a cotton plug or a filter. The filtrate was subjected to preparative HPLC (the preparative HPLC chart is shown in FIG. 1), and the fraction containing the collected ¹¹ C-labeled compound (d) was concentrated under reduced pressure using an evaporator. The concentrated solution was diluted with a clinical administration solution mainly composed of physiological saline and placed in a sterile vial. A part (20 μL) of this solution was subjected to analytical HPLC to identify the target compound, test the purity, and calculate the specific activity. The ¹¹ C-labeled compound (d) was identified using the unlabeled compound (d), and the following yields and analytical values are the results obtained from three experiments.

Total radioactivity of ¹¹ C-labeled compound (d): 3.8 ± 1.2 GBq, decay-corrected radiochemical yield based on [ ¹¹ C] methyl iodide: 63 ± 14%, synthesis time: 38 minutes, radiochemical purity: 99% or more, chemical purity: 99%, specific activity: 97 ± 10 GBq / μmol
Preparative conditions: Column preparative is made by Waters XBridge, 5μm, C18 10mm × 250mm , flow rate was 5 mL / min, mobile phase CH ₃ OH: 10 mM sodium phosphate buffer (pH = 7.4) = 50: 50 It was used. As a result of measurement with a UV detection wavelength of 254 nm and a γ-ray detector, the retention time of the ¹¹ C-labeled compound (d) was about 9.2 minutes.
Analysis conditions: Gemini NX, 5 μm, 4.6 mm × 150 mm manufactured by Phenomenex was used as the column, and analysis was performed at a column temperature of 30 ° C. The flow rate was 1 mL / min, and CH ₃ OH: 10 mM sodium phosphate buffer (pH = 7.4) = 50: 50 was used as the moving bed. As a result of measurement with a UV detection wavelength of 215 nm and a γ-ray detector, the radiochemical purity and chemical purity of the ¹¹ C-labeled compound (d) were both 99% or more.

<Experimental equipment, experimental method and reagents used>
As the labeling synthesizer, a standard automatic labeling synthesizer installed at RIKEN Molecular Imaging Science Research Center was used. A commercially available chemical was used as it was. Dehydrated N-methyl-2-pyrrolidinone (NMP) (Kanto Chemical Co., Inc.), Tris (dibenzylideneacetone) dipalladium (0) (Aldrich), tri-o-tolylphosphine (Aldrich), copper bromide (Made by Wako) and cesium fluoride (made by Wako) were used.

Example 4
<Synthesis of ¹¹ C-labeled isoquinoline derivative (d) 2>
Synthesis of (S) -hexahydro-2-methyl-1-[(4- [ ¹¹ C] methyl-5-isoquinolinyl) sulfonyl] -1H-1,4-diazepine Compound (d) from the compound described in Example 2 Can be obtained.
That is, ¹¹ C labeling by cross-coupling a boronic ester of an organic boronic acid compound with [ ¹¹ C] methyl iodide in the presence of a palladium complex, a phosphine ligand and a base in an aprotic polar solvent. An isoquinoline derivative (d) can be produced.

The following compounds of Examples 5 to 10 can be synthesized by using the methods described in Reference Example 1 and Examples 1 to 4.
Example 5
2,2,2-trifluoro-1- [4- {4- (tributylstannyl) isoquinolin-5-ylsulfonyl} -1,4-diazepan-1-yl] ethane
Example 6
2,2,2-trifluoro-1- [4- {4- (tributylstannyl) isoquinolin-5-ylsulfonyl} piperazin-1-yl] ethane
Example 7
(S) -2,2,2-trifluoro-N- (2- [3-methyl-4- {4- (tributylstannyl) isoquinolin-5-ylsulfonyl} -1,4-diazepan-1-yl ] -2-oxoethyl) acetamide
Example 8
5- (1,4-diazepan-1-ylsulfonyl) -4- [ ¹¹ C] methylisoquinoline
Example 9
4- [ ¹¹ C] methyl-5- (piperazin-1-ylsulfonyl) isoquinoline
Example 10
(S) -2-Amino-1- {3-methyl-4- (4- [ ¹¹ C] methylisoquinolin-5-ylsulfonyl) -1,4-diazepan-1-yl} ethanone

Example 11
<PET formulation example>
Preparation of intravenous PET preparation by dissolving the oily product of the compound (d) obtained by fractionation and concentration in Example 4 in, for example, a polysorbate 80-containing physiological saline and further diluting with physiological saline. Can do.

Example 12
<PET photography>
In order to perform PET imaging using the ¹¹ C-labeled isoquinoline derivative of the present invention as a PET probe, for example, the following operation may be performed.
That is, the ¹¹ C-labeled isoquinoline derivative (d) of the present invention is fractionated by HPLC to prepare the preparation described in Example 11, and this is administered as a PET probe to laboratory rats or the like by intravenous injection or the like. For example, continuous imaging is performed using a PET device for small animals (microPET, manufactured by SIMENS). Further, in order to examine the binding inhibitory effect of the non-RI-labeled substance, a solution obtained by diluting an unlabeled isoquinoline derivative with physiological saline is administered to a laboratory rat or the like by intravenous injection or the like. In addition, after PET imaging is completed, a tissue section is prepared, and an in vivo autoradiogram experiment is performed to examine its distribution. In this way, various PET images and their changes over time can be examined.

Claims (14)

¹¹ C-labeled isoquinoline derivative comprising general formula (1) or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

(Where
One of R ¹ and R ² represents a [ ¹¹ C] methyl group, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group, a nitro group, a fluoro group, or Represents an aryl group;
R ³ and R ⁴ each independently represent a hydrogen atom, alkyl group, alkenyl group, amino group, alkylamino group, dialkylamino group, aminoalkyl group, halogenoalkyl group, alkanoyl group, aminoalkanoyl group or alkylaminoalkanoyl group. Or R ³ and R ⁴ together form an alkylene group or an alkenylene group and are bridged between any two carbons in the cyclic diamino group in general formula (1) Yes;
R ⁵ represents a hydrogen atom, an aminoalkanoyl group or an alkylaminoalkanoyl group;
m represents a natural number of 1 to 4, and n represents 2 or 3. ) The ¹¹ C-labeled isoquinoline derivative according to claim 1, wherein R ¹ is a [ ¹¹ C] methyl group bonded to the 4-position of the isoquinoline ring. (S) -Hexahydro-2-methyl-1-[(4- [ ¹¹ C] methyl-5-isoquinolinyl) sulfonyl] -1H-1,4-diazepine, 5- (1,4-diazepan-1-ylsulfonyl) ) -4- [ ¹¹ C] methylisoquinoline, 4- [ ¹¹ C] methyl-5- (piperazin-1-ylsulfonyl) isoquinoline and (S) -2-amino-1- {3-methyl-4- (4) - ^[11 C] methyl-isoquinoline-5-ylsulfonyl) -1,4-diazepan-1-yl} compound selected from the group consisting ethanone or a pharmaceutically acceptable salt, hydrate, or solvate thereof Prodrug. A molecular probe for PET, comprising the ¹¹ C-labeled isoquinoline derivative according to claim 1. A compound for producing the ¹¹ C-labeled isoquinoline derivative according to claim 1, comprising the following general formula (2).

(Where
R ⁶ and R ⁷ either represents a trialkyltin group, a boronyl group or a dialkoxyboryl group, and the other represents a hydrogen atom, a cyano group, an alkyl group, a halogenoalkyl group, an alkenyl group, an alkoxy group, an alkylthio group, Represents a nitro group, a fluoro group or an aryl group;
R ³ and R ⁴ each independently represent a hydrogen atom, an alkyl group, an alkenyl group, an amino group, an alkylamino group, a dialkylamino group, an aminoalkyl group, a halogenoalkyl group, an alkanoyl group, an aminoalkanoyl group or an alkylaminoalkanoyl group. Or R ³ and R ⁴ together form an alkylene or alkenylene group and are bridged between any two carbons in the cyclic diamino group in general formula (2);
R ⁸ represents tert-butoxycarbonyl group, trihaloacetyl group, tert-butoxycarbonylaminoalkanoyl group, trihaloacetylaminoalkanoyl group, N-alkyl-N-tert-butoxycarbonylaminoalkanoyl group or N-alkyl-N-trihaloacetylamino. Represents an alkanoyl group;
m represents a natural number of 1 to 4, and n represents 2 or 3. ) R ⁶ is a trialkyltin group, boronyl group or dialkoxyboryl group bonded to the 4-position of the isoquinoline ring, and R ⁸ is a trihaloacetyl group, a trihaloacetylaminoalkanoyl group, or an N-alkyl-N-trihaloacetylaminoalkanoyl group 6. A compound according to claim 5, which is a group. R ⁶ is a trialkyltin group, boronyl group or dialkoxyboryl group bonded to the 4-position of the isoquinoline ring, R ³ is a hydrogen atom or a methyl group, R ⁴ and R ⁷ are hydrogen atoms, R ⁸ 6. The compound according to claim 5, wherein is a trihaloacetyl group, a trihaloacetylaminoalkanoyl group, or an N-alkyl-N-trihaloacetylaminoalkanoyl group, m is 1 or 2, and n is 2. Claim 1 wherein the isoquinoline ring of the compound according to any one of claims 5-7 by introducing a ^[11 C] methyl group, subsequent to, characterized in that to desorb R ⁸ under basic conditions The manufacturing method of the compound of any one of -3. Use of a compound according to any one of claims 5 to 7 for the preparation of an ¹¹ C labeled isoquinoline derivative according to any one of claims 1 to 3. A method for imaging a tissue, wherein the ¹¹ C-labeled isoquinoline derivative according to claim 1 is administered to a living body and the distribution of ¹¹ C in the living body is examined. The tissue imaging method according to claim 10, wherein the ¹¹ C distribution is examined by positrons.  The tissue imaging method according to claim 10, wherein the examination target is a brain. The tissue imaging method according to claim 10, wherein the distribution of protein kinase existing in a living body is examined by examining the distribution of ¹¹ C.  The tissue imaging method according to claim 13, wherein the protein kinase is Rho kinase.

Images