WO2014030709A1 - ミトコンドリアComplex-1の検出に適した化合物 - Google Patents
ミトコンドリアComplex-1の検出に適した化合物 Download PDFInfo
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- WO2014030709A1 WO2014030709A1 PCT/JP2013/072442 JP2013072442W WO2014030709A1 WO 2014030709 A1 WO2014030709 A1 WO 2014030709A1 JP 2013072442 W JP2013072442 W JP 2013072442W WO 2014030709 A1 WO2014030709 A1 WO 2014030709A1
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- 0 CC(C)(C)N(C1=O)N=CC(OCc(cc2)cnc2OCCOCC*)=C1Cl Chemical compound CC(C)(C)N(C1=O)N=CC(OCc(cc2)cnc2OCCOCC*)=C1Cl 0.000 description 2
- PNJYZWHRJKQPQT-UHFFFAOYSA-N CC(C)(C)N(C1=O)N=CC(OCc(cn2)ccc2OCCOCCO)=C1Cl Chemical compound CC(C)(C)N(C1=O)N=CC(OCc(cn2)ccc2OCCOCCO)=C1Cl PNJYZWHRJKQPQT-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/041—Heterocyclic compounds
- A61K51/044—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
- A61K51/0459—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with two nitrogen atoms as the only ring hetero atoms, e.g. piperazine
Definitions
- the present invention relates to a compound suitable for detection of mitochondrial Complex-1.
- PET method positron emission tomography
- SPECT method single photon tomography
- Diagnosis of various neuropsychiatric disorders including dementia is currently performed by evaluating the glucose metabolism of nerve cells by the PET method. At this time, 18 F-fluorodeoxyglucose ([ 18 F] FDG) is used as a probe for PET.
- 18 F-fluorodeoxyglucose [ 18 F] FDG
- [18 F] When evaluating the rat cerebral infarction model by PET method using FDG, in [18 F] ischemic disorders site of brain integration has been expected to decrease in FDG by the onset of cerebral infarction, expected On the other hand, it was revealed that [ 18 F] FDG accumulation increased. This indicates that [ 18 F] FDG is not suitable as a PET probe for evaluation after neuropathy.
- microglial cells that control immunity in the brain are activated, they accumulate at the site of ischemic injury where an inflammatory reaction is induced. Since this integrated microglia cells take up [18 F] FDG, that abnormal accumulation of [18 F] FDG in ischemic injury site of the brain is increased it has been confirmed.
- the present invention has been made in view of the above circumstances, and an object thereof is to provide a compound suitable for detection of mitochondrial Complex-1 that can be used as a labeling compound in the PET method.
- the present invention provides a compound represented by the formula (1-0).
- R represents —O (CH 2 ) n —, —O (CH 2 ) n OC 2 H 4 —, —CH 2 O (CH 2 ) n — or —CH 2 O ( CH 2 ) n OC 2 H 4 —, n represents an integer of 1 to 5, and Q 1 represents F or —OCH 3 )
- the compound represented by the formula (1-0) is a compound suitable for detection of mitochondrial Complex-1.
- Q 1 may be 18 F or —O 11 CH 3 .
- the compound can release positrons.
- the positron emitted from the compound immediately combines with the electron and emits ⁇ rays (annihilation radiation).
- ⁇ rays annihilation radiation
- the present invention provides a compound represented by formula (2-0).
- R represents —O (CH 2 ) n —, —O (CH 2 ) n OC 2 H 4 —, —CH 2 O (CH 2 ) n — or —CH 2 O ( CH 2 ) n OC 2 H 4 —
- n represents an integer of 1 to 5
- Q 2 represents a removable substituent (such as a substituted sulfonyloxy group, a halogen atom or a hydroxyl group).
- FIG. 3 is an NMR spectrum diagram of an intermediate of BMS-P. It is a graph which shows the binding affinity with respect to mitochondria Complex-1.
- FIG. 3 is an image showing the binding inhibitory effect of a PET probe by a specific inhibitor of mitochondrial Complex-1 in a rat brain section. It is a graph which shows the binding inhibitory effect of a PET probe by a mitochondrial Complex-1 specific inhibitor in a rat brain section.
- FIG. 2 is a PET image showing the binding inhibitory effect of a PET probe by a specific inhibitor of mitochondrial Complex-1 in the brain and heart of a living rat. 2 is a graph showing the binding inhibitory effect of a PET probe by a specific inhibitor of mitochondrial Complex-1 in the brain and heart of a living rat.
- FIG. 2 is a PET image showing the binding inhibitory effect of a PET probe by a specific inhibitor of mitochondrial Complex-1 in the brain of a living monkey.
- 2 is a graph showing the binding inhibitory effect of a PET probe by a specific inhibitor of mitochondrial Complex-1 in the brain of a living monkey.
- FIG. 6 is a PET image diagram and graph showing the correlation between [ 18 F] BCPP-EF binding and aging in the brain of a living monkey. It is the figure and graph of PET image which show the coupling
- a compound suitable for detection of mitochondrial Complex-1 is a compound represented by the formula (1-0) (hereinafter sometimes referred to as “compound (1-0)”).
- R is, -O (CH 2) n - , - O (CH 2) n OC 2 H 4 -, - CH 2 O (CH 2) n - or -CH 2 O (CH 2) n OC 2 H 4 - It is. n is an integer of 1 to 5, and preferably 2 to 4.
- Q 1 is F or —OCH 3 , and is preferably 18 F or —O 11 CH 3 . By setting Q 1 to —O 11 CH 3 or 18 F, the compound (1-0) can release a positron.
- Q 1 is —O 11 CH 3
- the half-life is as short as 20 minutes, so that it is possible to perform multiple measurements per day.
- Q 1 is 18 F
- the half-life is 110 minutes, which is longer than —O 11 CH 3 , so that one measurement time can be extended.
- the bonding position of —OCH 2 — bonded to the pyridazine ring and the bonding position of R in the pyridine ring are not particularly limited, but the bonding position of —OCH 2 — bonded to the pyridazine ring is the 5-position of the pyridine ring. It is preferable that the bonding position of R is the 2-position of the pyridine ring.
- the structural formula when the bonding position of —OCH 2 — bonded to the pyridazine ring is the 5-position of the pyridine ring and the bonding position of R is the 2-position of the pyridine ring is represented by the following formula (1-0 ′) Shown in
- a compound suitable for detection of mitochondrial Complex-1 is a compound represented by the formula (1) (hereinafter sometimes referred to as “compound (1)”). It is preferable.
- Compound (1) is less lipophilic than [ 18 F] BMS. Therefore, nonspecific binding of compound (1) is suppressed, and the binding specificity for mitochondrial Complex-1 tends to be higher than that of [ 18 F] BMS.
- a compound suitable for detection of mitochondrial Complex-1 may be a compound represented by the formula (1-2) (hereinafter referred to as “compound (1-2)”). .).
- compound (2-0) is a precursor of the compound (1-0).
- Q 2 is a detachable substituent (substituted sulfonyloxy group, halogen atom, hydroxyl group or the like).
- substituted sulfonyloxy group examples include a tosyloxy group (—OTs), a methanesulfonyloxy group (—OMs), a trifluoromethanesulfonyloxy group (—OTf), and a nitrobenzenesulfonyloxy group (—ONs).
- OTs are preferably used.
- halogen atom examples include fluorine, chlorine, bromine and iodine.
- the compound represented by the formula (2-0 ′) (hereinafter sometimes referred to as the compound (2-0 ′)) is a precursor of the compound (1-0 ′).
- the compound represented by the formula (2) (hereinafter sometimes referred to as the compound (2)) is a precursor of the compound (1).
- the compound represented by the formula (2-2) is a precursor of the compound (1-2).
- the compound (2-0) in which R is —CH 2 O (CH 2 ) n —, n is 2, and Q 2 is a hydroxyl group can be synthesized from known compounds. For example, the synthesis can be performed through the synthesis scheme (Steps 1 to 10) described in Experiment 2 of Examples described later.
- the compound (2-0) in which R is —CH 2 O (CH 2 ) n OC 2 H 4 — and Q 2 is a hydroxyl group is a synthetic scheme (steps 1 to 10) described in Experiment 2 of Examples described later. The synthesis is possible by referring to the synthesis scheme (c) described later.
- the compound (2) in which Q 2 is a hydroxyl group can be synthesized from a known compound.
- the synthesis can be performed via the synthesis schemes (a) to (h) described in Experiment 1 of Examples described later.
- the compound (2) in which Q 2 is a tosyloxy group can be synthesized from a known compound.
- the compound can be synthesized from the compound (2) in which Q 2 is a hydroxyl group via a synthesis scheme (i) described in Examples described later.
- the method for producing compound (1) wherein Q 1 is F from compound (2) is carried out by a method for fluorinating compound (2).
- a method for fluorinating compound (2) For example, when Q 2 is —OTs, the method for fluorinating the compound (2) is represented by the following synthesis scheme (A).
- Q 2 is another substituted sulfonyloxy group or a halogen atom, the corresponding compound (1) can be synthesized by the same synthesis scheme.
- the method for producing compound (1) wherein Q 1 is 18 F from compound (2) is carried out by a method of [ 18 F] fluorination of compound (2).
- a method of [ 18 F] fluorination of compound (2) is represented by the following synthesis scheme (B).
- Q 2 is another substituted sulfonyloxy group or a halogen atom
- the corresponding compound (1) can be synthesized by the same synthesis scheme.
- compound (2) is reacted with a complex of a macrocyclic ligand and [ 18 F] KF in a solvent to produce [ 18 F] fluorine. It is possible to
- the solvent for fluorination is not particularly limited as long as it can dissolve the starting material to some extent, and examples thereof include acetonitrile, dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and the like. Preferably used.
- macrocyclic ligands examples include 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo [8.8.8] hexacosane (K [2.2.2]), 1, 4,7,10,13,16-hexaoxacyclooctadecane (18-crown-6) and the like, and K [2.2.2] is preferably used.
- a method for producing a compound (1) in which Q 1 is —OCH 3 from a compound (2) in which Q 2 is a hydroxyl group is represented, for example, by the following synthesis scheme (C).
- a method for producing a compound (1) in which Q 1 is —O 11 CH 3 from a compound (2) in which Q 2 is a hydroxyl group is represented, for example, by the following synthesis scheme (D).
- 11 CH 3 OTf can be synthesized by a known method. For example, it can be synthesized via reaction formula (E).
- Compound (1) tends to specifically bind to mitochondrial Complex-1 when administered to a living body.
- Compound (1-2) tends to bind with high affinity to mitochondrial Complex-1 when administered to a living body. Therefore, compound (1-0) is suitable for detection of mitochondrial Complex-1. For example, if a fluorescent dye or the like is bound to compound (1-0), or positron labeling is performed on compound (1-0), it can be used as a labeled compound of mitochondrial Complex-1.
- Q 1 of the compound (1-0) is —O 11 CH 3 or 18 F
- the compound (1-0) can emit a positron.
- the positron emitted from the compound (1-0) immediately combines with the electron and emits ⁇ rays.
- the distribution of the compound (1-0) in the body can be quantitatively imaged over time. Therefore, by using compound (1-0), it is possible to detect a site where mitochondrial Complex-1 is present in the subject's living body and visualize the change over time.
- Compound (1-0) is useful as a mitochondrial Complex-1 detection reagent.
- the detection reagent contains the compound (1-0), and when administered to a living body, ⁇ -rays released from the compound (1-0) in the body are measured by the PET method, so that mitochondrial Complex-1 is present. Can be efficiently detected.
- the detection reagent is particularly suitable for the detection of mitochondrial Complex-1 in the brain.
- Compound (1-0) is also useful as a diagnostic agent for Parkinson's disease. According to the diagnostic agent, Parkinson's disease can be diagnosed by efficiently detecting a site where mitochondrial Complex-1 is present in the living body.
- the mitochondrial Complex-1 detection reagent and the Parkinson's disease diagnostic agent can be produced, for example, by dissolving the compound (1-0) in an arbitrary buffer solution.
- the detection reagent and the diagnostic agent are provided as a solution and may contain other components such as a surfactant, a preservative, and a stabilizer in addition to the buffer component.
- the administration method is usually intravenous administration.
- Examples of the mitochondrial Complex-1 detection reagent and the diagnostic agent for Parkinson's disease include, but are not limited to, humans, monkeys, mice, and rats.
- the measurement method is not particularly limited, and can be performed according to a known method.
- Silica Gel 60N (for flash chromatography) 40-50 ⁇ m manufactured by Kanto Chemical Co., Inc. was used as the silica gel used for silica gel column chromatography.
- Mucochloric acid 1 (50 g, 0.29 mol) was dissolved in water (440 mL), and sodium carbonate (15.3 g, 0.14 mol) was added. To the solution was added tert-butylhydrazine hydrochloride (36.9 g, 0.29 mol) at 0 ° C. The resulting reaction solution was stirred for 2.5 hours. The precipitated solid was filtered, washed with cold water, and then the precipitated solid was dried under reduced pressure to obtain Intermediate 2.
- Step 6 Compound 11 was synthesized according to synthesis scheme (f).
- Step 7 Compound 12 was synthesized according to the synthesis scheme (g).
- Process 8 Compound 13 was synthesized according to synthesis scheme (h).
- Step 9 Compound 14 was synthesized according to synthesis scheme (i).
- Step 10 Compound 15 (BCPP-EF) was synthesized according to synthesis scheme (j).
- Step 11 Compound 16 was synthesized according to synthesis scheme (k).
- a radioactivity peak with a retention time of 17.4 minutes was collected, and the collected solution was concentrated to dryness with an evaporator. Thereafter, the concentrated and dried product was redissolved in 0.1% Tween 80 / saline (5 mL) to recover [ 18 F] BCPP-EF (2.36 GBq).
- the proton was accelerated to 18 MeV in a cyclotron (HM-18, Sumitomo Heavy Industries, Tokyo). Meanwhile, a target gas in which pure nitrogen gas (G grade, Japan Fine Products, Kanagawa) was sealed at a pressure of approximately 17 kg / cm 2 was prepared. The target gas was irradiated with the proton at a current value of approximately 20 ⁇ A, and [ 11 C] was produced by 14 N (p, ⁇ ) 11 C nuclear reaction. Within the target gas, [ 11 C] was present in the chemical form [ 11 C] CO 2 .
- the precursor (compound 13, 2 mg) was dissolved in 2-butanone (Wako Pure Chemical Industries, Tokyo, 0.3 mL). It was prepared by adding NaH (about 2 mg) to the obtained precursor solution. The [ 11 C] methyl triflate that had been distilled was introduced into the resulting precursor solution, and the introduction of the [ 11 C] methyl triflate was stopped when the radioactivity reached equilibrium. Then, the methylation reaction of the precursor was performed on the conditions for 5 minutes at 40 degreeC.
- the production amount of [ 11 C] BCPP-EM was 0.26 to 2.01 GBq, and the radiochemical purity was 93.6% or more.
- the retention time of the precursor of [ 11 C] BCPP-EM was 5.2 minutes, and the retention time of [ 11 C] BCPP-EM was 10 minutes.
- the retention time of [ 11 C] BCPP-EM in analytical HPLC was 4.0 minutes.
- the target compound was extracted with ethyl acetate (1 L ⁇ 2).
- the organic layer was washed successively with water (1 L) and saturated brine (1 L). After drying the organic layer with magnesium sulfate, the solvent of the organic layer was distilled off under reduced pressure to obtain Compound 9 ′ (16.12 g, yield 66.0%) as a pale tan solid.
- the obtained filtrate was washed with saturated brine (300 mL) and dried over sodium sulfate. Thereafter, the solvent of the filtrate was distilled off under reduced pressure.
- Step 7 Under an Ar atmosphere, sodium hydride (2.430 g, 60.74 mmol as 60% equivalent) was added separately to a dioxane solution (100 mL) of compound 18 ′ (10.76 g, 51.91 mmol) under ice cooling, The reaction was stirred at room temperature for 15 minutes. Subsequently, a dioxane solution (60 mL) of compound 5b (18.24 g, 60.74 mmol) was added, and the reaction solution was stirred at 50 ° C. for 2 hours. Ice water (300 mL) was added to the reaction solution, and the target compound was extracted with ethyl acetate (200 mL ⁇ 2).
- Step 9 Under an Ar atmosphere, a THF solution (200 mL) of compound 4 (3.185 g, 15.72 mmol), compound 11 ′′ (4.201 g, 15.72 mmol) and triphenylphosphine (6.185 g, 23.58 mmol) was added. Under ice cooling, a THF solution (23 mL) of diisopropyl azodicarboxylate (DIAD, 4.768 g, 23.58 mmol) was added dropwise, and the reaction solution was stirred at room temperature for 16 hours.
- DIAD diisopropyl azodicarboxylate
- Step 11 To a solution of compound 13 ′′ (140.6 mg, 0.3822 mmol), triethylamine (386.7 mg, 3.822 mmol) and 4-dimethylaminopyridine (4.67 mg, 0.03822 mmol) in methylene chloride (3 mL) was added ⁇ 10 P-Toluenesulfonyl chloride (109.3 mg, 0.5734 mmol) was added at ° C. After adding p-toluenesulfonyl chloride, the reaction solution was stirred at ⁇ 10 ° C. for 16 hours. Ice water (20 mL) was poured into the reaction solution, and the target compound was extracted with methylene chloride (20 mL).
- One compound 15 ′′ (BMS-P) (47.6 mg, yield 25.4%) was obtained.
- a dose-response curve of a test substance and a positive substance is prepared, and the concentration at which the test substance and the positive substance inhibit the binding between the tracer and the mitochondrial Complex-1 (receptor) by 50% (IC 50 value) and absolute inhibition constant (Ki).
- test substances BMS, BCPP-EF, BCPP-BF, BCPP-PF, BCPP-EM and BMS-P were used. Details are shown below.
- the mitochondrial fraction stock solution was diluted with a buffer solution to prepare a mitochondrial fraction solution having a concentration twice the final concentration (prepared when used).
- the final concentration of the mitochondrial fraction solution was 45 ⁇ g protein / mL.
- test substance solution A solution having a concentration 100 times the final concentration was prepared by serially diluting the test substance with DMSO. Further, a test substance solution having a concentration 10 times the final concentration was prepared by diluting the prepared solution of each concentration 10 times with Milli-Q water (preparation before use).
- the final concentration of the test substance was in the following concentration range: BMS: 3 ⁇ 10 ⁇ 11 to 3 ⁇ 10 ⁇ 8 mol / L BCPP-EF: 1 ⁇ 10 ⁇ 10 to 1 ⁇ 10 ⁇ 7 mol / L BCPP-BF: 3 ⁇ 10 ⁇ 11 to 3 ⁇ 10 ⁇ 8 mol / L BCPP-PF: 1 ⁇ 10 ⁇ 10 to 1 ⁇ 10 ⁇ 7 mol / L BCPP-EM; 3 ⁇ 10 ⁇ 10 to 3 ⁇ 10 ⁇ 7 mol / L BMS-P: 1 ⁇ 10 ⁇ 7 to 1 ⁇ 10 ⁇ 4 mol / L.
- Positive substances were weighed and dissolved in DMSO.
- a solution having a concentration 100 times the final concentration was prepared by serial dilution of the resulting solution with DMSO. Furthermore, the solution of each concentration prepared was diluted 10-fold with Milli-Q water to prepare a positive substance solution having a concentration 10 times the final concentration (prepared at the time of use). Rotenone was used as a positive substance.
- the final concentration of the positive substance was 3 ⁇ 10 ⁇ 11 to 3 ⁇ 10 ⁇ 8 mol / L.
- substitution substance solution A substitution substance was weighed and dissolved in DMSO to prepare a solution having a concentration 100 times the final concentration. Further, the prepared solution was diluted 10-fold with Milli-Q water to prepare a substitution substance solution having a concentration 10 times the final concentration (prepared at the time of use). Rotenone was used as the substitute substance. The final concentration of the substitution substance was 1 ⁇ 10 ⁇ 5 mol / L.
- tracer solution having a concentration 10 times the final concentration was prepared by diluting the tracer stock solution with a buffer solution (preparation at the time of use). Tracer, was used Dihydrorotenone [2-isopropyl- 3 H ( N)]. The final concentration of the tracer was 4.5 nmol / L for the first time, 4.4 nmol / L for the second time, and 4.5 nmol / L for the third time.
- Measurement procedure Measurement was carried out in the following order. Two sample samples were prepared at each concentration and measured three times. 1: To the tube for calculating non-specific binding, 100 ⁇ L of a displacement substance solution was added (final concentration of DMSO: 1%). To the tube for calculating total binding, 100 ⁇ L of 10% DMSO was added (final concentration of DMSO: 1%). 100 ⁇ L each of the test substance solution and the positive substance solution was added to the tube for calculating the inhibition rate of the test substance or positive substance (final concentration of DMSO: 1%). 2: Buffer solution (300 ⁇ L) was added to each tube. 3: Tracer solution (100 ⁇ L) was added to each tube. 4: Mitochondrial fraction solution (500 ⁇ L) was added to each tube.
- reaction solution contained in each tube was incubated at 22 ° C. for 30 minutes.
- 6 The reaction solution was filtered through a cell harvester (GF / C, Whatman), and the filtered filter paper was washed three times with 50 mmol / L Tris-HCl buffer (pH, 7.40, 3 mL).
- 7 The filter paper was transferred to a vial for measurement, a liquid scintillator (PICO-FLUOR TM PLUS, 5 mL) was added, and the amount of radioactivity was measured with a liquid scintillation counter (measurement time, 2 minutes).
- the inhibition rate was calculated by the formula of "100-binding rate". Coupling rate: [(B ⁇ N) / (B 0 ⁇ N)] ⁇ 100 (%) B: Bound radioactivity in the presence of test substance (individual value) B 0 : Total bound radioactivity in the absence of test substance (average value) N: Non-specific binding radioactivity (average value) The inhibition rate was calculated as 0% when the inhibition rate was 0% or less, and as 100% when the inhibition rate exceeded 100%. For the positive substance, the inhibition rate was calculated in the same manner as the test substance.
- Dose-response curve creation (IC 50 value calculation)
- the dose-response curve is the ratio ((BN) / (Bn) of the specific binding activity (BN) in the presence of the test substance to the total binding activity (B 0 -N) in the absence. 0- N)) was converted to a logit and then applied to a logit-log model that plots against the common logarithm of the final concentration of the test substance. The following regression equation was used for regression of dose-response curve.
- Rats were anesthetized with chloral hydrate (400 mg / kg; IP) and decapitated. Thereafter, the brain was immediately sampled and cooled, and immersed in a sherbet-like ACSF (artificial cerebrospinal fluid). Necessary blocks of the obtained brain were cut out and adhered to a fixed base of a vibration microtome HM650V (Thermo Fisher Scientific, Waltham, USA). Thereafter, brain slices having a thickness of 300 ⁇ m were prepared. The excised brain slices were soaked in 24 ml ACSF bubbled with O 2 for 30 minutes to calm the cell activity.
- ACSF artificial cerebrospinal fluid
- the OHP sheet on which the brain slices were arranged was placed on an imaging plate (ST-VI, Fuji Film, Tokyo) having sensitivity to radiation and allowed to stand (contact) for about 1 hour in a dark place. .
- the imaging plate was detected with a fluoro image analyzer FLA-7000 (Fuji Film, Tokyo).
- FLA-7000 Fluji Film, Tokyo.
- FIGS. It was found that [ 18 F] BCPP-EF is more susceptible to the binding inhibition effect by rotenone than [ 18 F] BMS. This suggests that [ 18 F] BCPP-EF is more specific for mitochondrial Complex-1 than [ 18 F] BMS.
- a similar experiment was performed on [ 18 F] BCPP-BF, [ 18 F] BCPP-PF, and [ 18 F] BCPP-EM, and it was confirmed that it was affected by the binding inhibition effect by rotenone.
- a cephalic vein or saphenous vein was secured as a route for intravenous administration of the test animal, and a femoral artery or posterior tibial artery was secured as a route for collecting arterial blood, respectively.
- the amount of radioactivity of [ 18 F] BMS, [ 18 F] BCPP-EF or [ 11 C] BCPP-EM was measured.
- the entire dose was administered intravenously over about 30 seconds.
- emission measurement (10 seconds 6 frames, 30 seconds 6 frames, 1 minute 12 frames, 3 minutes 25 frames, 5 minutes 6 frames, 121 minutes, 55 frames) was started.
- FIG. 10 shows the PET measurement results of young monkeys (5 years old) and old monkeys (22 years old).
- [ 18 F] BCPP-EF was used as a PET probe. It was found that young monkeys accumulated more PET probes in the brain than older monkeys. This suggests that aging reduces the activity of mitochondrial Complex-1 in the brain.
Abstract
Description
Rが-CH2O(CH2)n-であり、nが2であり、Q2が水酸基である化合物(2-0)は、公知の化合物から合成可能である。例えば、後述する実施例の実験2に記載の合成スキーム(工程1~10)を経て合成が可能である。Rが-CH2O(CH2)nOC2H4-でありQ2が水酸基である化合物(2-0)は、後述する実施例の実験2に記載の合成スキーム(工程1~10)及び後述する合成スキーム(c)を参照することで、合成が可能である。
化合物5a(10.00g、68.41mmol)、トリエチルアミン(10.38g、102.6mmol)及び4-ジメチルアミノピリジン(836mg、6.841mmol)の塩化メチレン溶液(100mL)に、氷冷下でp-トルエンスルホニルクロライド(13.69g、71.83mmol)を分けて添加した。添加後、氷冷下で反応液を16時間撹拌した。反応液に氷水(200mL)を注ぎ込み、塩化メチレン(100mL)で、目的化合物を抽出した。有機層を飽和食塩水(100mL)で洗浄し、硫酸マグネシウムで乾燥した。その後、有機層の溶媒を減圧留去した。得られた残渣を中圧分取(シリカゲル120g、ヘキサン/酢酸エチル=5/1~1/1)で精製し、無色液体である化合物5b(18.38g、収率89.4%)を得た。
Ar雰囲気下、塩化カルシウム(29.92g、269.6mmol)、エタノール(390mL)及びTHF(390mL)を、反応器に仕込み、続いて氷冷下、水素化ホウ素ナトリウム(20.40g、539.2mmol)を分割して反応器に投入した。水素化ホウ素ナトリウムを投入した後、氷冷下で反応液を2.5時間撹拌した。反応液に氷冷下、化合物7’(30.09g、134.8mmol)を分割して投入し、氷冷下で反応液を30分間攪拌した。反応液を氷水(1.5L)に注ぎ込んだ後、塩化アンモニウム(300g)を撹拌しながら反応液に加えた。目的化合物を酢酸エチル(1L×2)で抽出した。有機層を水(1L)、飽和食塩水(1L)で順次洗浄した。硫酸マグネシウムで有機層を乾燥した後、有機層の溶媒を減圧留去し、淡黄褐色固体である化合物9’(16.12g、収率66.0%)を得た。
化合物9’(17.07g、94.21mmol)及びp-トルエンスルホン酸一水和物(18.11g、94.21mmol)の塩化メチレン溶液(500mL)に、氷冷下、3,4-ジヒドロ-2H-ピラン(23.55mL、259.2mmol)を加え、室温で反応液を18時間攪拌した。有機層を飽和重曹水(500mL)、飽和食塩水(500mL)で順次洗浄した。硫酸マグネシウムで有機層を乾燥した後、有機層の溶媒を減圧留去することで、淡黄褐色液体である化合物10’’’(42.72g、粗収率170.9%)を得た。化合物10’’’はクルードのまま次工程に供した。
Ar雰囲気下、水素化アルミニウムリチウム(7.988g、210.5mmol)のTHF懸濁液(100mL)に、クルードの化合物10’’’(42.72g、Net=24.99g,94.21mmol)のTHF溶液(200mL)を氷冷下で滴下した。その後、室温で反応液を1時間攪拌した。氷冷下、反応液に水(8mL)、15%水酸化ナトリウム水溶液(8mL)、水(24mL)を順次滴下して反応を停止した。得られた混合物をろ過し、スラリーを酢酸エチル(600mL)で洗浄した。得られたろ液を飽和食塩水(300mL)で洗浄、硫酸ナトリウムで乾燥した。その後、ろ液の溶媒を減圧留去した。得られた残渣を中圧分取(シリカゲル200g、ヘキサン/酢酸エチル=1/1~酢酸エチルonly)で精製し、淡黄褐色液体である化合物16’(14.98g、化合物9’からの収率71.2%)を得た。
Ar雰囲気下、化合物16’(14.98g、67.09mmol)のDMF溶液(60mL)に氷冷下、水素化ナトリウム(3.489g、60%換算として87.22mmol)を分けて添加し、室温で反応液を1時間攪拌した。続いて化合物8(10.53g、100.7mmol)を、反応液に加え、50℃で3時間攪拌した。反応液に氷水(300mL)を加え、酢酸エチル(300mL)で目的化合物を抽出した。有機層を水(300mL)、飽和食塩水(300mL)で順次洗浄した。硫酸マグネシウムで有機層を乾燥した後、有機層の溶媒を減圧留去した。得られた残渣を中圧分取(シリカゲル200g、ヘキサン/酢酸エチル=10/1~1/1)で精製し、淡黄褐色液体である化合物17’(15.40g、収率78.8%)を得た。
化合物17’(15.40g、52.85mmol)のメタノール溶液(154mL)に、p-トルエンスルホン酸一水和物(502.7mg、2.643mmol)を加え、室温で反応液を18時間攪拌した。その後、反応液を8時間加熱還流した。反応液を濃縮し、得られた残渣を中圧分取(シリカゲル200g、ヘキサン/酢酸エチル=1/1~酢酸エチルonly)で精製し、淡黄褐色液体である化合物18’(10.78g、収率98.4%)を得た。
Ar雰囲気下、化合物18’(10.76g、51.91mmol)のジオキサン溶液(100mL)に、氷冷下、水素化ナトリウム(2.430g、60%換算として60.74mmol)を分けて添加し、室温で反応液を15分攪拌した。続いて化合物5b(18.24g、60.74mmol)のジオキサン溶液(60mL)を加え、50℃で反応液を2時間攪拌した。反応液に氷水(300mL)を加え、酢酸エチル(200mL×2)で目的化合物を抽出した。有機層を飽和食塩水(200mL)で洗浄した。有機層を硫酸ナトリウムで乾燥した後、有機層の溶媒を減圧留去した。得られた残渣を中圧分取(シリカゲル200g、ヘキサン/酢酸エチル=4/1~酢酸エチルonly)で精製し、淡黄褐色液体である化合物19(14.21g、収率81.6%)を得た。
Ar雰囲気下、化合物19(14.21g、43.67mmol)のDMSO溶液(375mL)に、カリウムt-ブトキシド(49.00g、436.7mmol)を添加し、60℃で反応液を30分間攪拌した。反応液を氷水中(1L)に注ぎ込み、酢酸エチル(500mL×4)で目的化合物を抽出した。有機層を飽和食塩水(500mL)で洗浄し、硫酸ナトリウムで乾燥した。その後、有機層の溶媒を減圧留去した。得られた残渣を中圧分取(シリカゲル200g、ヘキサン/酢酸エチル=1/1~酢酸エチルonly)で精製し、淡黄色液体である化合物11’’、(4.201g、収率37.1%)を得た。
Ar雰囲気下、化合物4(3.185g、15.72mmol)、化合物11’’(4.201g、15.72mmol)及びトリフェニルホスフィン(6.185g、23.58mmol)のTHF溶液(200mL)に、氷冷下、アゾジカルボン酸ジイソプロピル(DIAD、4.768g、23.58mmol)のTHF溶液(23mL)を滴下し、室温で反応液を16時間攪拌した。反応液を濃縮し、得られた残渣を中圧分取(シリカゲル200g、ヘキサン/酢酸エチル=4/1~1/1、2回)で精製し、微黄色液体である化合物12’’、(431mg、収率6.1%)を得た。
化合物12’’(430mg、0.9515mmol)のメタノール溶液(4.3mL)に、p-トルエンスルホン酸一水和物(9.05mg、0.0457mmol)を加え、室温で反応液を18時間攪拌した。反応液を更に60℃で4時間撹拌した。反応液を濃縮し、得られた残渣を中圧分取(シリカゲル60g、ヘキサン/酢酸エチル=3/1~酢酸エチルonly)で精製し、無色液体である化合物13’’、(242mg、収率69.2%)を得た。化合物13’’のNMRスペクトルを図1に示す。
化合物13’’(140.6mg、0.3822mmol)、トリエチルアミン(386.7mg、3.822mmol)及び4-ジメチルアミノピリジン(4.67mg、0.03822mmol)の塩化メチレン溶液(3mL)に、-10℃でp-トルエンスルホニルクロライド(109.3mg、0.5734mmol)を添加した。p-トルエンスルホニルクロライドを添加した後、-10℃で反応液を16時間撹拌した。反応液に氷水(20mL)を注ぎ込み、塩化メチレン(20mL)で目的化合物を抽出した。有機層を飽和食塩水(10mL)で洗浄し、硫酸マグネシウムで乾燥した。その後、有機層の溶媒を減圧留去し、赤褐色で粘稠な液体である化合物14’’(201.2mg、粗収率100.8%)を得た。化合物14’’は、クルードのまま直ちに次工程に供した。
クルードな化合物14’’(195mg、Net=193.5mg,0.3706mmol)をTHF(2mL)に溶解し、1MのTBAF/THF(2mL)、TBAF・xH2O(500mg)を加え、室温で反応液を2時間攪拌した。反応液に氷水(50mL)を注ぎ込み、酢酸エチル(50mL)で目的化合物を抽出した。有機層を飽和食塩水(20mL)で洗浄し、硫酸マグネシウムで乾燥した。その後、有機層の溶媒を減圧留去した。得られた残渣を中圧分取(シリカゲル60g、ヘキサン/酢酸エチル=4/1~1/1)で精製し、化合物13’’(50mg)から別途調製及び精製したロットとあわせて赤褐色液体である化合物15’’(BMS-P)(47.6mg、収率25.4%)を得た。
被験物質及び陽性物質のdose-response curveを作成し,被験物質及び陽性物質がトレーサーとミトコンドリアComplex-1(レセプター)との結合を50%抑制する濃度(IC50値)及び絶対阻害定数(Ki)を算出した。被験物質としては、BMS、BCPP-EF、BCPP-BF、BCPP-PF、BCPP-EM及びBMS-Pを用いた。詳細を以下に示す。
動物組織を秤量(湿重量)し、2倍量のホモジネート緩衝液(250mmol/Lのスクロース、1mmol/Lのコハク酸及び0.2mmol/LのEDTAの10mmol/LのTris-HCl緩衝液(pH、7.40))を加え、氷冷下でホモジナイズした。その後、ホモジナイズした懸濁液を遠心分離した(1200×g、4°C、20分間)。上清を採取し、遠心分離した(26000×g、4°C、15分間)。沈殿している残渣を採取し、100mg tissue eq./mLとなるようにホモジネート緩衝液を加えホモジナイズした。得られたミトコンドリア画分原液は使用時まで-80°Cに保存した。ミトコンドリア画分のタンパク質濃度はBCA Protein Assay Reagent(PIERCE社製)を用いて測定した。動物組織は、ラットの脳及びウシの心臓を用いた。
上記ミトコンドリア画分原液を緩衝液で希釈することによって、最終濃度の2倍濃度のミトコンドリア画分溶液を調製した(用時調製)。ミトコンドリア画分溶液の最終濃度は、45μg protein/mLとした。
被験物質をDMSOで段階希釈することによって、最終濃度の100倍濃度の溶液を調製した。さらに、調製した各濃度の溶液を、Milli-Q水で10倍希釈することによって最終濃度の10倍濃度の被験物質溶液を調製した(用時調製)。
被験物質の最終濃度は、以下の濃度範囲とした:
BMS ;3×10-11~3×10-8mol/L
BCPP-EF ;1×10-10~1×10-7mol/L
BCPP-BF ;3×10-11~3×10-8mol/L
BCPP-PF ;1×10-10~1×10-7mol/L
BCPP-EM ;3×10-10~3×10-7mol/L
BMS―P ;1×10-7~1×10-4mol/L。
陽性物質を秤量し、DMSOで溶解した。得られた溶液をDMSOで段階希釈することによって、最終濃度の100倍濃度の溶液を調製した。さらに、調製した各濃度の溶液を、Milli-Q水で10倍希釈することによって、最終濃度の10倍濃度の陽性物質溶液を調製した(用時調製)。陽性物質は、ロテノンを用いた。陽性物質の最終濃度は、3×10-11~3×10-8mol/Lとした。
置換物質を秤量し、DMSOで溶解することによって、最終濃度の100倍濃度の溶液を調製した。さらに、調製した溶液を、Milli-Q水で10倍希釈することによって最終濃度の10倍濃度の置換物質溶液を調製した(用時調製)。置換物質は、ロテノンを用いた。置換物質の最終濃度は、1×10-5mol/Lとした。
トレーサー原液を緩衝液で希釈することによって、最終濃度の10倍濃度のトレーサー溶液を調製した(用時調製)。トレーサーは、Dihydrorotenone [2-isopropyl-3H(N)]を用いた。
トレーサーの最終濃度は、1回目が4.5nmol/Lであり、2回目が4.4nmol/Lであり、3回目が4.5nmol/Lであった。
以下の順序に従い測定を実施した。各濃度において2例サンプルを調製し、それぞれ3回測定した。
1:非特異的結合を算出するためのチューブには、置換物質溶液を100μL添加した(DMSOの最終濃度:1%)。総結合を算出するためのチューブには、10%DMSOを100μL添加した(DMSOの最終濃度:1%)。被験物質又は陽性物質の阻害率を算出するためのチューブには、被験物質溶液又は陽性物質溶液を、それぞれ100μL添加した(DMSOの最終濃度:1%)。
2:各チューブに緩衝液(300μL)を添加した。
3:各チューブにトレーサー溶液(100μL)を添加した。
4:各チューブにミトコンドリア画分溶液(500μL)を添加した。
5:各チューブ含まれる反応液を22°Cで30分間インキュベートした。
6:反応液をセルハーベスターによって濾過(GF/C、Whatman)し、濾過した濾紙を50mmol/LのTris-HCl緩衝液(pH、7.40、3mL)で3回洗浄した。
7:測定するためのバイアルビンに、濾紙を移し、液体シンチレーター(PICO-FLUORTM PLUS、5mL)を添加し、液体シンチレーションカウンターで、放射能量を測定(測定時間、2分間)した。
阻害率は、「100-結合率」の式によって算出した。
結合率:[(B-N)/(B0-N)]×100(%)
B :被験物質存在下での結合放射能量(個別値)
B0 :被験物質非存在下での総結合放射能量(平均値)
N :非特異的結合放射能量(平均値)
阻害率が0%以下の場合には0%として、100%を超えた場合には100%として阻害率を算出した。
陽性物質に関しても被験物質と同様に阻害率を算出した。
Dose-response curveは、被験物質存在下での特異的結合放射能(B-N)と非存在下での総結合放射能(B0-N)との比((B-N)/(B0-N))をlogit変換した後、被験物質の最終濃度の常用対数値に対してプロットするlogit-logモデルにあてはめて作成した。
Dose-response curveの回帰は、次の回帰式を用いた。
Y=aX+b
(Y=logit y=ln(y/(1-y))、y=(B-N)/(B0-N))
(X=log x、xは被験物質の最終濃度を示す。)
(a=定数、b=定数)
得られた回帰式から、IC50値を算出した。回帰の際、被験物質の最終濃度における阻害率平均が5%~95%の範囲を超えたものについては採用せず、範囲内の連続して増加する阻害率を用いてIC50値を算出することとした。
IC50値の算出の試験に用いたトレーサー濃度(L)、得られたIC50値及び、Scatchard解析の試験により求めたトレーサーのミトコンドリアComplex-1に対するKd値を用いて、次式より被験物質及び陽性物質のKi値を算出した。
ラットを抱水クロラール(400mg/kg;I.P.)で麻酔し断頭した。その後、直ちに脳をサンプリングしてよく冷やし、シャーベット状のACSF(人工脳脊髄液)に浸けた。得られた脳の必要なブロックを切り出し、バイブレーションミクロトームHM650V(サーモフィッシャーサイエンティフィック、ウォルサム、米国)の固定台に接着した。その後、300μm厚の脳のスライスを作製した。切り出された脳のスライスは、O2でバブリングしている24mlのACSFに30分間浸けて細胞の活動を落ち着かせた。その後、ACSFに、ベヒクル(Vehicle)又はロテノン(Rotenone、各ビーカーに2nM、20nM、200nM、20000nMの濃度になるよう調整した。)を20μL加え30分間バブリングした。さらに、ACSFに、[18F]BMS又は[18F]BCPP-EFを1MBq/ml加え30分間バブリングを続けた。バブリング終了後、脳のスライスを取り出し、100mlの新しいACSFに移した。その後、更に30分間、ACSFをバブリングすることによって余分なRIを洗浄した。その後、OHPシート上に上記脳のスライスを並べ、スキャナー(EPSON:GT-600)で画像を撮影した。その後、上記脳のスライスが並べられたOHPシートを、放射線に対して感度を有するイメージングプレート(ST-VI、富士フィルム、東京)の上に置いて、暗所で1時間程度放置(コンタクト)した。上記イメージングプレートはフルオロ・イメージアナライザーFLA-7000(富士フィルム、東京)で検出を行った。結果を図3及び4に示す。[18F]BCPP-EFの方が、[18F]BMSよりも、ロテノンによる結合阻害効果の影響を受けやすいことが分かった。このことは、[18F]BCPP-EFの方が、[18F]BMSよりもミトコンドリアComplex-1の特異性が高いことを示唆している。[18F]BCPP-BF、[18F]BCPP-PF及び[18F]BCPP-EMについても同様の実験を行ったところ、ロテノンによる結合阻害効果の影響を受けることが確認された。
ラットを抱水クロラール(400mg/kg;I.P.)で麻酔し、計測用ベッドに固定した。その直後、ロテノン(0.1mg/kg/h)又はベヒクルを上記ラットの尾静脈から投与した。投与中に脳の位置を同定するためにClairvivo-CT(島津製作所)による計測を実施した。その後、Clairvivo PETへ計測用ベッドを移動した。ロテノン(0.1mg/kg/h)又はベヒクルの投与が終了した後、直ちに上記ラットの尾静脈から[18F]BMS又は[18F]BCPP-EFを約8MBq/匹でボーラス投与し、60分間計測を行った。計測中は上記ラットの尾静脈から抱水クロラール(100mg/kg/hr)持続投与することでラットを非動化させた。上記ラットの小脳、大脳皮質、線条体及び心臓において、ロテノンによる結合阻害効果を観察した。その結果を図5及び6に示す。いずれの組織においても、[18F]BCPP-EFの方が、[18F]BMSよりも、ロテノンの投与によって、取り込み量が減少していることが分かった。
光増感反応を利用してヒトと同様に実際に血栓で中大脳動脈(middle cerebral artery :MCA)を閉塞したラットの脳梗塞(Photochemically induced thrombosis :PIT)モデルを用いて実験を行った。ラットは4%ハロセン(30%O2、70%Room Air)にて麻酔導入し、術中は1.5~2%のハロセン濃度で維持した。手術用顕微鏡下で左眼窩縁に沿って皮膚を切開し、頭骸底を歯科用ドリルで左側中大脳動脈が硬膜下に見えるように約3mmの楕円形の窓(穴)を削開した。光増感剤であるローズベンガル(20mg/kg)を上記ラットの尾静脈から投与し、540nmの波長の緑色光を上記中大脳動脈に10分間照射することで、ラットに脳梗塞を発症させた。
サルの脳内におけるロテノン投与によるPETプローブの結合阻害実験、老化に伴う脳内のミトコンドリアComplex-1の活性の変化、及び、パーキンソン病モデルのサルの脳内におけるミトコンドリアComplex-1の活性の評価を行った。いずれの実験においてもPET計測は以下の方法によって行った。
サルの脳内におけるロテノン投与によるPETプローブの結合阻害実験では、ロテノンは0.1mg/kg/hrとなるように静脈内注射によって被験動物に投与した。その後、プローブとして[18F]BMS、[18F]BCPP-EF又は[11C]BCPP-EMを投与してPET計測した。結果を図8及び9に示す。[18F]BCPP-EF及び[11C]BCPP-EMの方が、[18F]BMSよりも、ロテノンによる結合阻害効果の影響を受けやすいことが分かった。サルの場合でも[18F]BCPP-EF及び[11C]BCPP-EMの方が、[18F]BMSよりもミトコンドリアComplex-1の特異性が高いことが分かった。
若齢サル(5歳)と老齢サル(22歳)とのPET計測結果を図10に示す。PETプローブとして[18F]BCPP-EFを用いた。若齢サルの方が老齢サルよりもPETプローブが多く脳内に集積していることが分かった。このことは、老化によって脳内のミトコンドリアComplex-1の活性が減少していることを示唆している。
正常なサルとパーキンソン病モデルのサルとの脳内におけるミトコンドリアComplex-1の活性をPET計測によって比較した。PETプローブとして[18F]BCPP-EFを用いた。パーキンソン病モデルのサルは、以下の様にして準備した。雄の若齢カニクイザル(体重3~6kg)に、ドパミン神経を選択的に障害するMPTP(1-メチル-4-フェニル-1,2,3,6-テトラヒドロピリジン)を毎週1回のペースで6ヶ月間に渡り、MPTPの総量として15~20mg投与した。MPTPの投与にあたっては、上記カニクイザルの健康状態を確認しながら行った。上記カニクイザルが餌をつまみ上げる様子を観察することで、四肢の運動機能障害の度合いを評価すると同時に、PET計測を用いてドパミン再吸収部位の低下を計測することによって、最終的にパーキンソン病状態である事を確認した。結果を図11に示す。パーキンソン病モデルのサルでは、正常なサルに比べて、脳内に集積しているPETプローブの量が少ないことが分かった。このことは、パーキンソン病の発症とミトコンドリアComplex-1の活性とに相関関係があることを示唆している。
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Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015056521A1 (ja) * | 2013-10-16 | 2015-04-23 | 浜松ホトニクス株式会社 | 癌に対する治療効果の診断剤 |
JP2017206457A (ja) * | 2016-05-18 | 2017-11-24 | 浜松ホトニクス株式会社 | 肝臓及び/又は腎臓機能の診断剤 |
JP2018199626A (ja) * | 2017-05-25 | 2018-12-20 | 浜松ホトニクス株式会社 | 褐色脂肪細胞の活性評価用プローブ |
JP2019127447A (ja) * | 2018-01-23 | 2019-08-01 | 浜松ホトニクス株式会社 | 脳神経機能異常検出剤 |
WO2022014605A1 (ja) | 2020-07-14 | 2022-01-20 | 浜松ホトニクス株式会社 | 膵臓機能の診断剤 |
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WO2022145260A1 (ja) * | 2020-12-28 | 2022-07-07 | 浜松ホトニクス株式会社 | 被験者の腎臓以外の臓器及び組織のミトコンドリア機能を評価する方法 |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61243078A (ja) * | 1985-04-19 | 1986-10-29 | Nissan Chem Ind Ltd | ピリダジノン誘導体,その製造法および殺虫・殺ダニ・殺線虫・殺菌剤 |
JPS62207262A (ja) * | 1986-03-07 | 1987-09-11 | Nissan Chem Ind Ltd | ピリダジノン誘導体、その製法および殺虫、殺ダニ、殺線虫、殺菌剤 |
JPS63159373A (ja) * | 1986-12-24 | 1988-07-02 | Nissan Chem Ind Ltd | ピリダジノン類および殺虫、殺ダニ、殺線虫剤 |
JPH07252236A (ja) * | 1994-01-28 | 1995-10-03 | Takeda Chem Ind Ltd | 抗癌剤 |
JP2006199615A (ja) * | 2005-01-19 | 2006-08-03 | Astellas Pharma Inc | 眼機能検査薬およびそれを用いた眼機能測定方法 |
WO2008023780A1 (fr) * | 2006-08-25 | 2008-02-28 | Gifu University | Procédé de méthylation rapide, coffret pour préparer un traceur pet et procédé de fabrication d'un traceur pet |
JP2011513306A (ja) | 2008-02-29 | 2011-04-28 | ランセウス メディカル イメージング, インコーポレイテッド | 灌流造影を含む適用のための造影剤 |
CN102336741A (zh) * | 2011-07-06 | 2012-02-01 | 北京师范大学 | 氟-18标记的心肌灌注显像剂及其制备方法和应用 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4837217A (en) * | 1985-04-19 | 1989-06-06 | Nissan Chemical Industries, Ltd. | Pyridazinone derivatives, preparation thereof, and insecticidal, acaricidal, nematicidal, fungicidal compositions |
EP0665223A1 (en) * | 1994-01-28 | 1995-08-02 | Takeda Chemical Industries, Ltd. | Antitumor agent, novel 3(2H)-pyridazinone derivatives and their preparation |
US7344702B2 (en) | 2004-02-13 | 2008-03-18 | Bristol-Myers Squibb Pharma Company | Contrast agents for myocardial perfusion imaging |
AU2013203000B9 (en) * | 2012-08-10 | 2017-02-02 | Lantheus Medical Imaging, Inc. | Compositions, methods, and systems for the synthesis and use of imaging agents |
-
2013
- 2013-08-22 WO PCT/JP2013/072442 patent/WO2014030709A1/ja active Application Filing
- 2013-08-22 JP JP2014531670A patent/JP6223981B2/ja active Active
- 2013-08-22 CN CN201380044392.5A patent/CN104583192B/zh active Active
- 2013-08-22 US US14/422,834 patent/US9598399B2/en active Active
- 2013-08-22 EP EP13831121.2A patent/EP2915809B1/en active Active
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61243078A (ja) * | 1985-04-19 | 1986-10-29 | Nissan Chem Ind Ltd | ピリダジノン誘導体,その製造法および殺虫・殺ダニ・殺線虫・殺菌剤 |
JPS62207262A (ja) * | 1986-03-07 | 1987-09-11 | Nissan Chem Ind Ltd | ピリダジノン誘導体、その製法および殺虫、殺ダニ、殺線虫、殺菌剤 |
JPS63159373A (ja) * | 1986-12-24 | 1988-07-02 | Nissan Chem Ind Ltd | ピリダジノン類および殺虫、殺ダニ、殺線虫剤 |
JPH07252236A (ja) * | 1994-01-28 | 1995-10-03 | Takeda Chem Ind Ltd | 抗癌剤 |
JP2006199615A (ja) * | 2005-01-19 | 2006-08-03 | Astellas Pharma Inc | 眼機能検査薬およびそれを用いた眼機能測定方法 |
WO2008023780A1 (fr) * | 2006-08-25 | 2008-02-28 | Gifu University | Procédé de méthylation rapide, coffret pour préparer un traceur pet et procédé de fabrication d'un traceur pet |
JP2011513306A (ja) | 2008-02-29 | 2011-04-28 | ランセウス メディカル イメージング, インコーポレイテッド | 灌流造影を含む適用のための造影剤 |
CN102336741A (zh) * | 2011-07-06 | 2012-02-01 | 北京师范大学 | 氟-18标记的心肌灌注显像剂及其制备方法和应用 |
Non-Patent Citations (3)
Title |
---|
HUISMAN ET AL., J. NUCL. MED, vol. 49, pages 630 - 636 |
See also references of EP2915809A4 * |
YALAMANCHILI, P. ET AL., J. NUCL. CARDIOL, vol. 14, 2007, pages 782 - 788 |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015056521A1 (ja) * | 2013-10-16 | 2015-04-23 | 浜松ホトニクス株式会社 | 癌に対する治療効果の診断剤 |
US9872928B2 (en) | 2013-10-16 | 2018-01-23 | Hamamatsu Photonics K.K. | Diagnostic agent for therapeutic effect on cancer |
JP2017206457A (ja) * | 2016-05-18 | 2017-11-24 | 浜松ホトニクス株式会社 | 肝臓及び/又は腎臓機能の診断剤 |
JP7005126B2 (ja) | 2016-05-18 | 2022-02-04 | 浜松ホトニクス株式会社 | 肝臓及び/又は腎臓機能の診断剤 |
JP2018199626A (ja) * | 2017-05-25 | 2018-12-20 | 浜松ホトニクス株式会社 | 褐色脂肪細胞の活性評価用プローブ |
JP7007111B2 (ja) | 2017-05-25 | 2022-01-24 | 浜松ホトニクス株式会社 | 褐色脂肪細胞の活性評価用プローブ |
JP2019127447A (ja) * | 2018-01-23 | 2019-08-01 | 浜松ホトニクス株式会社 | 脳神経機能異常検出剤 |
JP7134630B2 (ja) | 2018-01-23 | 2022-09-12 | 浜松ホトニクス株式会社 | 脳神経機能異常検出剤 |
WO2022014605A1 (ja) | 2020-07-14 | 2022-01-20 | 浜松ホトニクス株式会社 | 膵臓機能の診断剤 |
WO2022092215A1 (ja) | 2020-10-29 | 2022-05-05 | 浜松ホトニクス株式会社 | 組織・臓器の相互作用の検出剤 |
WO2022145260A1 (ja) * | 2020-12-28 | 2022-07-07 | 浜松ホトニクス株式会社 | 被験者の腎臓以外の臓器及び組織のミトコンドリア機能を評価する方法 |
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US20150225368A1 (en) | 2015-08-13 |
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