Organic compound and application thereof in preparation of 18kDa-targeted translocation protein PET probe and medicine

Abstract

The invention relates to the technical field of targeted drugs, in particular to an organic compound and application thereof in preparation of a PET probe targeting 18kDa translocation protein and pharmacy. The organic compound has a structure as shown in formula (I), M1 and M2 are respectively and independently selected from D or H, and X1 is selected from F18 or F. Compared with a traditional PET probe, the organic compound has the TSPO targeting characteristic, can be used for preparing the TSPO targeting PET probe, is good in stability in in-vivo plasma metabolism and lower in non-specific uptake, and can be used for diagnosis, scientific research and medicine preparation of neuritis.

Description

Technical Field

[0001] The present application relates to the technical field of targeted drugs, and in particular to an organic compound and its application in preparing a PET probe and a drug targeting an 18kDa translocation protein. Background Art

[0002] Neuroinflammation is an inflammatory response caused by pathological damage such as infection, trauma or toxin accumulation in the central nervous system. It is one of the important pathophysiological processes of many central nervous system diseases. In this process, glial cells, especially microglia, play a vital role. Microglia are natural immune cells of the central nervous system. They can recognize changes in death or injury signals in the central nervous system, proliferate and aggregate at the site of lesions, and play a role in monitoring the microenvironment of the central nervous system. Under the action of pathological factors, microglia can be rapidly activated into reactive microglia, which not only have the function of protecting neurons, but also can secrete cytotoxic factors and complement proteins, thereby damaging neurons.

[0003] TSPO (18kDa translocator protein), a translocator protein widely distributed throughout the body and located in the outer membrane of mitochondria, is closely related to mitochondrial function. When inflammation occurs in the central nervous system, the expression of TSPO in activated microglia increases significantly, so TSPO can also be considered a potential biomarker for neuroinflammation. The development of PET probes targeting TSPO targets has become a hot spot for the application of PET in the field of the central nervous system. 11 As the first PET probe targeting TSPO, C-PK11195 has been applied to monitor inflammation in a variety of neurological diseases. 11 The short half-life of C 11 The low target-to-target ratio of C-PK11195 itself limits its widespread clinical application.

[0004] In order to overcome these limitations, current research directions are devoted to developing PET probes with higher target-to-specimen ratios, more stable pharmacokinetics, and more accurate visualization of TSPO expression. Many second-generation TSPO-PET probes have been gradually applied in clinical and scientific research, effectively improving the target-to-specimen ratio during imaging. However, due to the single nucleotide polymorphisms of the TSPO gene itself, these probes have different abilities to bind to TSPO. In order to solve this problem, designing PET probes that are insensitive to TSPO gene polymorphisms will help to carry out clinical trials with larger samples. Summary of the invention

[0005] Based on this, one or more embodiments of the present application provide an organic compound having a structure as shown in formula (I):

[0006]

[0007] in,

[0008] M1 and M2 are each independently selected from D or H, and at least one of M1 and M2 is D;

[0009] X1 from F 18 or F.

[0010] This organic compound can target the 18kDa translocator protein (TSPO) with high selectivity and is insensitive to TSPO gene polymorphism. At the same time, it has more stable plasma metabolism in the body and has lower background uptake, and can be well applied to the visualization of neuroinflammation in the brain.

[0011] Preferably, M1 and M2 are both D.

[0012] The present application also provides a method for preparing the organic compound described above, comprising the following steps:

[0013]

[0014] Compound A is reacted with a deuterated reducing agent to reduce the ester group and deuterate the hydrogen at the ortho position to obtain compound B;

[0015] The compound B is subjected to oxidation and esterification reaction to obtain compound C;

[0016] The compound C is ring-opened and subjected to an amidation reaction with the compound D to obtain the compound E;

[0017] The OH of the compound E is subjected to a halogenation reaction with an X2 donor reagent to obtain a compound F, wherein X2 is selected from Br or Cl;

[0018] X2 of the compound F is subjected to a halogen exchange reaction with a donor reagent X1 to prepare an organic compound having a structure as shown in formula (I).

[0019] Further, the deuterated reducing agent includes at least one of deuterated lithium aluminum hydride and deuterated sodium borohydride; and / or

[0020] The molar ratio of the compound A to the deuterated reducing agent is 1:(3.5-4.5).

[0021] Further, the compound B is subjected to oxidation and esterification reaction, comprising:

[0022] Mix the compound B and the oxidant at 15°C±1°C, and carry out reduction and esterification reaction at 25°C-35°C for 25-35 minutes;

[0023] Optionally, the oxidant is selected from chromate pyridinium salt oxidants;

[0024] Optionally, the molar ratio of the compound B to the oxidant is 1:(2-3).

[0025] Further, the compound C is ring-opened and subjected to an amidation reaction with the compound D, comprising:

[0026] Add Lewis acid to the organic solution of compound D at 15°C±1°C, and stir at 25°C-35°C for 10-20 minutes to obtain a first mixed solution;

[0027] Adding the compound C to the first mixed solution, and subjecting to thermal reflux reaction for 24 hours to 48 hours;

[0028] Optionally, the molar ratio of the compound C to the compound D is 1:(4-4.5);

[0029] Optionally, the molar ratio of the Lewis acid to the compound D is 1:(3-3.2).

[0030] Optionally, the temperature of the halogen exchange reaction is 110°C±5°C, and the time is 8min to 12min.

[0031] The present application also provides a PET probe targeting 18kDa translocation protein, which comprises the organic compound described above.

[0032] The present application also provides a use of the organic compound described above in the preparation of a drug for treating neuritis.

[0033] Furthermore, the organic compound treats neuritis by targeting the 18kDa translocator protein.

[0034] Compared with conventional PET probes targeting TSPO, the organic compounds of the present application exhibit higher stability and lower nonspecific uptake in in vivo plasma metabolism, and can be effectively used for visualization of neuroinflammation in the brain.

[0035] The preparation method of the organic compound of the present application can be efficiently synthesized on an automated synthesis module, the method is highly operable and feasible, and has important application value in the research and diagnosis of central nervous system neuroinflammatory related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0037] Figure 1 Prepared for an embodiment of this application 18 F-Reaction scheme of d1-LW223 precursor and d1-LW223 standard.

[0038] Figure 2 Prepared for an embodiment of this application 18 F-Reaction scheme of d2-LW223 precursor and d2-LW223 standard.

[0039] Figure 3 For one embodiment of this application, HPLC separation and purification was used 18 Chromatogram obtained for F-d1-LW223.

[0040] Figure 4 For one embodiment of this application, HPLC separation and purification was used 18 Chromatogram obtained for F-d1-LW223.

[0041] Figure 5 For the detection of an embodiment of this application 18 HPLC chromatogram of F-d1-LW223 purity.

[0042] Figure 6 This is an embodiment of the present application 18 HPLC chromatogram of co-injection detection of F-d1-LW223 and d1-LW223.

[0043] Figure 7 For one embodiment of this application, HPLC separation and purification was used 18 Chromatogram obtained for F-d2-LW223.

[0044] Figure 8 For one embodiment of this application, HPLC separation and purification was used 18 Chromatogram obtained for F-d2-LW223.

[0045] Fig. 9 For the detection of an embodiment of this application 18 HPLC chromatogram of F-d2-LW223 purity.

[0046] Fig.10 This is an embodiment of the present application 18HPLC chromatogram of co-injection detection of F-d2-LW223 and d2-LW223.

[0047] Fig.11 is a compound of one embodiment of the present application 18 F-d2-LW223, 18 F-d1-LW223 and 18 Comparative results of F-LW223 metabolism in cynomolgus monkeys, Fig.11 Medium A is 60 minutes 18 F-d2-LW223, 18 F-d1-LW223 and 18 Results of plasma metabolism analysis of F-LW223 in cynomolgus monkeys. Fig.11 B is 18 F-d2-LW223, 18 F-d1-LW223 and 18 Whole-body static imaging results of F-LW223 in cynomolgus monkeys.

[0048] Fig.12 For an embodiment of the present application 18 Representative PET images of F-d2-LW223 in the brain of stroke rats, Fig.12 A in the middle is the baseline A, Fig.12 Middle B is the result of PK11195 pretreatment, where the blue area represents the infarct area and the red area represents the healthy contralateral side. Fig.12 C in the middle is the brain of the rat model of infarction 18 The AUC analysis results of the F-d2-LW223PET imaging time radioactivity curve from 40 to 60 minutes. Fig.12 Middle D shows the immunofluorescence results of TSPO and iBa1 in the damaged area of ​​striatum of stroke rats (×100).

[0049] Fig.13 For an embodiment of this application, 18 F-d2-LW223PET evaluates the therapeutic effect of anti-CNS inflammatory drug edaravone and dextran on stroke patients. Fig.13 A representative before and after treatment 18 F-d2-LW223 PET images, the upper two images (treatment group) are PET images before and after treatment with edaravone and dextran, and the lower two images (control group) are PET images before and after treatment with control intervention measures. Fig.13 Middle B is a diagram showing the changes in SUVr before and after treatment for the 7 individuals currently included in the study.

[0050] Fig.14 For an embodiment of this application, 18F-d2-LW223PET evaluates the neuronal status of the brain in a 13-month-old Huntington mouse model. Fig.14 A is a representative image of the whole brain of a 13-month-old Huntington mouse model 18 F-d2-LW223 PET image, Fig.14 Middle B is a comparison between the Huntington mouse group and the wild-type mouse group using a t-test to test SUV uptake in different brain regions. The results showed that Huntington mice had increased uptake in the striatum, thalamus, hippocampus, and cortex.

[0051] Fig.15 For an embodiment of this application, 18 F-d2-LW223PET evaluates the neural verification in the brain of Huntington mouse model at different time periods; Fig.15 A in the middle is a representative image of the whole brain of Huntington's mouse model at 1.5 months, 3 months, 7 months, 10 months, and 13 months old 18 F-d2-LW223PET imaging; Fig.15 B shows the changes of various brain regions of Huntington mice over time. 18 F-d2-LW223PET uptake curve;

[0052] Fig.16 For an embodiment of this application, 18 F-d2-LW223PET evaluates the neural verification in the brain of the Huntington mouse model at different time periods, showing the immunofluorescence staining results of TSPO (pink) and Iba-1 (red) in the striatum of 140QHDKI mice. DETAILED DESCRIPTION

[0053] The present application will be further described below in conjunction with the embodiments and examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. In addition, it should be understood that after reading the content taught in the present application, those skilled in the art may make various changes or modifications to the present application, and these equivalent forms also fall within the protection scope of the claims attached to the present application.

[0054] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0055] the term

[0056] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0057] The term "and / or" used in this article includes any one of two or more related listed items, and also includes any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0058] Herein, "preferred", "preferable", "better", etc. are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of this application.

[0059] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0060] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0061] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0062] The temperature parameters in this application, unless otherwise specified, are allowed to be either constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±0.5°C, ±0.4°C, ±0.3°C, ±0.2°C, and ±0.1°C are allowed.

[0063] In the present application, weight may be μg, mg, g, kg or other mass units known in the chemical industry.

[0064] In the present application, unless otherwise specified, the size, particle size and diameter generally refer to the average value.

[0065] The present application provides an organic compound having a structure as shown in formula (I):

[0066]

[0067] in,

[0068] M1 and M2 are each independently selected from D or H, and at least one of M1 and M2 is D;

[0069] X1 from F 18 or F.

[0070] This organic compound can target the 18kDa translocator protein (TSPO) with high selectivity and is insensitive to TSPO gene polymorphism. At the same time, it has more stable plasma metabolism in the body and has lower background uptake, and can be well applied to the visualization of neuroinflammation in the brain.

[0071] Preferably, M1 and M2 are both D.

[0072] Compared with conventional PET probes targeting TSPO, the organic compounds of the present application exhibit higher stability and lower nonspecific uptake in in vivo plasma metabolism, and can be effectively used for visualization of neuroinflammation in the brain.

[0073] The present application also provides a method for preparing the above-mentioned organic compound. The method for preparing the organic compound of the present application can be efficiently synthesized on an automated synthesis module. The method is highly operable and feasible, and has important application value in the research and diagnosis of central nervous system neuroinflammatory related diseases.

[0074] The present application also provides a method for preparing the above organic compound, comprising the following steps:

[0075]

[0076]

[0077] Compound A is reacted with a deuterated reducing agent to reduce the ester group and deuterate the hydrogen at the ortho position to obtain compound B;

[0078] The compound B is subjected to oxidation and esterification reaction to obtain the compound C;

[0079] Compound C is ring-opened and subjected to amidation reaction with compound D to obtain compound E;

[0080] The OH of compound E is subjected to a halogenation reaction with an X2 donor reagent to prepare compound F, wherein X2 is selected from Br or Cl;

[0081] X2 of compound F is subjected to a halogen exchange reaction with a donor reagent X1 to prepare an organic compound having a structure as shown in formula (I).

[0082] Optionally, the X2 donor reagent includes thionyl chloride, phosphorus oxychloride, N-halosuccinimide, dihalogenated hydantoin, trihaloisocyanuric acid, 1,2-dihalogenated ethane, pyrophosphoryl chloride, cuprous chloride, etc., where halogen refers to Br or Cl.

[0083] Alternatively, the X1 donor reagent includes tetrabutylammonium fluoride (TBAF), tetra-n-butylammonium difluorotriphenylsilicate (TBAT), and alkali metal fluorides such as KF or CsF.

[0084] Further, the deuterated reducing agent includes at least one of deuterated lithium aluminum hydride and deuterated sodium borohydride; and / or

[0085] The molar ratio of compound A to the deuterated reducing agent is 1:(3.5-4.5).

[0086] Further, compound B is subjected to oxidation and esterification reaction, comprising:

[0087] Mix compound B and an oxidant at 15°C ± 1°C, and perform reduction and esterification reaction at 25°C to 35°C for 25 to 35 minutes;

[0088] Optionally, the oxidant is selected from chromate pyridinium salt oxidants;

[0089] Optionally, the molar ratio of compound B to the oxidant is 1:(2-3).

[0090] Further, the compound C is ring-opened and subjected to an amidation reaction with the compound D, comprising:

[0091] Add Lewis acid to the organic solution of compound D at 15°C ± 1°C, and stir at 25°C to 35°C for 10 to 20 minutes to obtain a first mixed solution;

[0092] Add compound C to the first mixed solution and reflux for 24 to 48 hours;

[0093] Optionally, the molar ratio of compound C to compound D is 1:(4-4.5);

[0094] Optionally, the molar ratio of Lewis acid to compound D is 1:(3-3.2).

[0095] Optionally, the temperature of the halogen exchange reaction is 110°C ± 5°C, and the time is 8 min to 12 min.

[0096] Lewis acids include aluminum chloride (AlCl3), boron trifluoride (BF3), aluminum trichloride (AlCl3), trimethylaluminum (Al(CH3)3), antimony pentafluoride (SbF5), sulfur trioxide (SO3), iron bromide (FeBr3), niobium pentachloride (NbCl5) and trifluoromethanesulfonates of lanthanide elements.

[0097] In one embodiment of the present application, the Lewis acid is selected from trimethylaluminum (Al(CH3)3), which can better carry out the ring-opening reaction of compound C and the amidation reaction with compound D.

[0098] The present application also provides the use of the above organic compound in preparing a PET probe targeting 18kDa translocation protein.

[0099] The present application also provides the use of the above organic compound in preparing a drug for treating neuritis.

[0100] Furthermore, the organic compounds treat neuritis by targeting the 18kDa translocator protein.

[0101] The following are some specific embodiments.

[0102] For experimental parameters not specified in the following specific embodiments, reference is made to the instructions given in the present application document, and reference may also be made to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by manufacturers.

[0103] The raw materials and reagents involved in the following specific examples can be obtained from commercial sources, or can be prepared by those skilled in the art according to known methods.

[0104] 1. Preparation and characterization of compounds

[0105] refer to Figure 1 and Figure 2 The synthetic route shown is 18 F-d1-LW223 precursor and 18 F-d1-LW223 precursor and d1-LW223 and d2-LW223 used as controls.

[0106] 1. Preparation 18F-d1-LW223 precursor

[0107] Compound 1 (1.50 g, 5.74 mmol, 1.00 eq) was added to anhydrous THF (30.0 ml) and cooled to 0°C. Then LiAlH4 (0.123 g, 0.5 eq) and LiAlD4 (0.123 g, 0.5 eq) were slowly added at 0°C, and the reaction mixture was stirred at room temperature (25°C) for 1 hour. TLC (dichloromethane / methanol=20 / 1, compound 1R f =0.4, compound 2R f =0.14) indicated that the reaction was complete. The reaction was quenched with 1.0 ml of water, 1.0 ml of 15% aqueous sodium hydroxide solution, and 3.0 ml of water at 0°C. The resulting mixture was filtered, diluted with dichloromethane (20 ml), washed with water (20 ml x 3) and saturated aqueous sodium chloride solution (20.0 ml), and dried over sodium persulfate. The solution was concentrated under reduced pressure to obtain crude product 5 (about 1.00 g), which was used directly in the next step without further purification.

[0108] In dichloromethane (36.0 ml), the crude product 2 (1.80 g, 6.73 mmol, 1.00 eq) was dissolved, PCC (3.63 g, 16.8 mmol, 2.50 eq) was added at 15°C and stirred at room temperature (25°C) for 0.5 h (the reaction time should not be too long and the temperature should not be too high). TLC (dichloromethane / methanol = 20 / 1, compound 2R f =0.14, compound 3R f =0.58) showed that a small amount of compound 2 was not reacted. The reaction mixture was filtered and concentrated under reduced pressure to obtain a crude product 3 (about 1.50 g), which was used directly in the next step without purification.

[0109] In dichloromethane (11.4 ml), compound 4 (353 mg, 4.05 mmol, 1.40 eq) was dissolved under N2 atmosphere, AlMe3 (1.88 ml, 1.30 eq, 2M in toluene) was slowly added at 15°C, and stirred at room temperature (25°C) for 15 minutes. Then a solution of compound 3 (760 mg, 2.89 mmol, 1.00 eq, 7.60 ml dichloromethane) was added and heated to reflux for 2 days. TLC (petroleum ether / ethyl acetate = 1 / 1, compound 3R f =0.58, compound 5R f =0.39) indicated that the reaction was complete. The reaction was quenched with 1M hydrochloric acid (20.0 mL), extracted with dichloromethane (30.0 mL×3), dried over sodium persulfate, filtered and concentrated under reduced pressure to give crude product 5 (456 mg), which was used directly in the next step without purification.

[0110] A dichloromethane solution of compound 5 (600 mg, 1.71 mmol, 1.00 eq; dissolved in 60.0 mL of DCM) was prepared, and dichlorothionyl (SOCl2, 4.42 g, 2.70 mL, 22 eq) was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give a crude product. The crude product was diluted with a saturated aqueous sodium bicarbonate solution (30.0 mL) and extracted with dichloromethane (20.0 mL×3). The organic phases were combined, washed with a saturated aqueous sodium chloride solution (20.0 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a crude mixture, which was obtained by silica gel column chromatography (PE / EtOAc=1 / 0-10 / 1-5 / 1) 18 F-d1-LW223 precursor (358 mg, four-step synthesis total yield 12.1%, light yellow solid).

[0111] High-resolution detection results: [M+H] + :C 22 H 23 DClN2O.368.9031, Found:368.8404.

[0112] 2. Preparation 18 F-d2-LW223 precursor

[0113] Compound 1 (1.50 g, 5.74 mmol, 1.00 eq) was added to anhydrous THF (30.0 ml) and cooled to 0°C. Then LiAlD4 (0.981 g, 23.4 mmol, 4.00 eq) was slowly added at 0°C, and the reaction mixture was stirred at room temperature (25°C) for 1 hour. TLC (dichloromethane / methanol=20 / 1, compound 1R f =0.4, compound 2R f =0.15) indicated that the reaction was complete. The reaction was quenched with 1.0 ml of water, 1.0 ml of 15% aqueous sodium hydroxide solution, and 3.0 ml of water at 0°C. The resulting mixture was filtered, diluted with dichloromethane (20 ml), washed with water (20 ml x 3) and saturated aqueous sodium chloride solution (20.0 ml), and dried over sodium persulfate. The solution was concentrated under reduced pressure to give a crude product 5 (about 1.00 g), which was used directly in the next step without further purification.

[0114] In dichloromethane (36.0 ml), the crude product 2 (1.80 g, 6.73 mmol, 1.00 eq) was dissolved, PCC (3.63 g, 16.8 mmol, 2.50 eq) was added at 15°C and stirred at room temperature (25°C) for 0.5 h (the reaction time should not be too long and the temperature should not be too high). TLC (dichloromethane / methanol = 20 / 1, compound 2R f =0.15, compound 3Rf =0.6) showed that a small amount of compound 2 was not reacted. The reaction mixture was filtered and concentrated under reduced pressure to obtain a crude product 3 (about 1.50 g), which was used directly in the next step without purification.

[0115] In dichloromethane (11.4 ml), compound 4 (353 mg, 4.05 mmol, 1.40 eq) was dissolved under N2 atmosphere, AlMe3 (1.88 ml, 1.30 eq, 2M in toluene) was slowly added at 15°C, and stirred at room temperature (25°C) for 15 minutes. Then a solution of compound 3 (760 mg, 2.89 mmol, 1.00 eq, 7.60 ml dichloromethane) was added and heated to reflux for 2 days. TLC (petroleum ether / ethyl acetate = 1 / 1, compound 3R f =0.6, compound 5R f =0.4) indicated that the reaction was complete. The reaction was quenched with 1M hydrochloric acid (20.0 mL), extracted with dichloromethane (30.0 mL x 3), dried over sodium persulfate, filtered and concentrated under reduced pressure to give crude product 5 (456 mg), which was used directly in the next step without purification.

[0116] A dichloromethane solution of compound 5 (600 mg, 1.71 mmol, 1.00 eq; dissolved in 60.0 mL of DCM) was prepared, and dichlorothionyl (SOCl2, 4.42 g, 2.70 mL, 22 eq) was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give a crude product. The crude product was diluted with a saturated aqueous sodium bicarbonate solution (30.0 mL) and extracted with dichloromethane (20.0 mL×3). The organic phases were combined, washed with a saturated aqueous sodium chloride solution (20.0 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a crude mixture, which was obtained by silica gel column chromatography (PE / EtOAc=1 / 0-10 / 1-5 / 1) 18 F-d2-LW223 precursor (590 mg, four-step synthesis total yield 19.8%, light yellow solid).

[0117] 3. Preparation 18 F-d1-LW223

[0118] 3.1 Automated labeling of drugs using All in One equipment

[0119] (1) Drying method for obtaining fluorine-18 anion

[0120] Fluorine-18 anions are generated by bombarding H2 with a proton beam in a medical cyclotron. 18O is produced and transmitted to the All in One device through a pipeline. Waters Sep-Paklight QMA solid phase extraction columns (Tianjin Dexiang Maolong Technology Co., Ltd., 186004051, SEP-PAK LIGHT QMA50BX) were activated with sodium bicarbonate aqueous solution (7.3%) and sterile water for injection, respectively, and then used to capture and separate H2 18 Fluorine-18 anions in O. 1 mL of TEAB eluent (6 mg TEAB in 0.7 mL ACN and 0.3 mL H2O) was automatically drawn with a 3 mL syringe to elute the solid phase extraction column, and the eluent entered the reaction bottle.

[0121] The reaction flask was heated to 85°C and blown with dry nitrogen gas for 5 min. Then 1 mL of acetonitrile was added to the reaction flask and the mixture was evaporated to dryness at 85°C for 5 min and 110°C for 2 min. 18 F-TEAF / TEAB mixture. The reaction flask was then cooled to 40°C.

[0122] (2) Fluorine-18 anion and 18 F-d1-LW223 precursor reaction

[0123] 4.0mg 18 The F-d1-LW223 precursor (dissolved in 1.0 mL ACN) was added to the reaction bottle, and the reaction bottle was heated to 110°C for 10 min. The reaction bottle was then cooled to 40°C, and 1.5 mL ACN + 1.0 mL H2O were added to quench the reaction.

[0124] (3) HPLC separation and purification of radioactive products

[0125] 3.5 mL of solution was drawn from the reaction bottle and injected into the loop. In order to reduce the loss of the product in the reaction bottle, 3 mL of acetonitrile was drawn and injected into the reaction bottle again, and then sucked out and injected into the loop again.

[0126] The parameters of HPLC separation and purification are as follows:

[0127] Instrument: Shimadzu HPLC LC-16P;

[0128] Detector: Japan Shimadzu UV detector (SPD-16, λ=254nm) and radioactivity detector;

[0129] Semi-preparative column: OSAKA SODA CAPCELL PAK C18, 5 μm, 10 mm ID × 250 mm;

[0130] Column temperature: 20°C;

[0131] Mobile phase solution: CH3CN / H2O=70 / 30;

[0132] Flow rate: 3 ml per minute;

[0133] The UPLC detection chromatogram is as follows: Figure 3 , Figure 3 The product retention time is 17 min.

[0134] 3.2 Automated labeling of drugs using the GE Tracerlab FX2N device

[0135] (1) Fluorine-18 anion drying method

[0136] Fluorine-18 anions are generated by bombarding H2 with a proton beam in a medical cyclotron. 18 O was produced and transferred to the GE Tracerlab FX2N device through a pipeline. A Waters Sep-Pak light QMA solid phase extraction column (Tianjin Dexiang Maolong Technology Co., Ltd., 186004051, SEP-PAK LIGHTQMA 50BX) was activated with sodium bicarbonate aqueous solution (7.3%) and sterile water for injection, respectively, and then used to capture and separate H2 18 Fluorine-18 anion in O. Elution with alkaline solution in V6 (6 mg TEAB + 0.7 mL anhydrous ethanol + 0.3 mL sterile water) 18 F - To reaction bottle No. 2, evaporate to dryness at 65°C for 3 min, add the liquid in V7 (1 mL acetonitrile) to reaction bottle No. 2, evaporate to dryness at 65°C and 95°C for 10 min. Then cool the reaction bottle to 50°C.

[0137] (2) Fluorine-18 anion and 18 F-d1-LW223 precursor reaction

[0138] 4.0mg 18 The F-d1-LW223 precursor was dissolved in 1.0 mL ACN in V8, added to the reaction bottle No. 2, and heated to 110°C for 10 min. Then the reaction bottle No. 2 was cooled to 40°C, and V9 (1.5 mL acetonitrile + 1 mL water) was added to the reaction bottle 2 and transferred to Tube 2 to quench the reaction.

[0139] (3) HPLC separation and purification of radioactive products

[0140] Draw 3.5 mL of product from Tube 2 and insert it into the loop.

[0141] The parameters of HPLC separation and purification are as follows:

[0142] Instrument: Shimadzu HPLC LC-16P;

[0143] Detector: Japan Shimadzu UV detector (SPD-16, λ=245nm) and radioactivity detector;

[0144] Semi-preparative column: OSAKA SODACAPCELL PAK C18, 5 μm, 10 mm ID × 250 mm;

[0145] Column temperature: 20°C;

[0146] Mobile phase solution: CH3CN / H2O=70 / 30;

[0147] Flow rate: 3 ml per minute;

[0148] The UPLC detection chromatogram is as follows: Figure 4 , Figure 4 The retention time of the intermediate product is 17 min.

[0149] 3.3 Radioactive product purity detection

[0150] The parameters for purity testing are as follows:

[0151] Instrument: Shimadzu HPLC LC-16P;

[0152] Detector: Japan Shimadzu UV detector (SPD-16, λ=245nm) and radioactivity detector;

[0153] Analytical column: GL Sciences, WondaSil C18-WR, 4.6 × 150 mm;

[0154] Column temperature: 20°C;

[0155] Mobile phase solution: CH3CN / H2O=70 / 30;

[0156] Flow rate: 1 mL / min;

[0157] Test results such as Figure 5 , Figure 5 The retention time of the radioactive product and the standard compound was 6.54min.

[0158] 3.4 Product consistency testing

[0159] The radioactive product 18 F-d1-LW223 and standard compound d1-LW223 were co-injected and tested, and the test parameters were as follows3:

[0160] Instrument: Shimadzu HPLC LC-16P;

[0161] Detector: Japan Shimadzu UV detector (SPD-16, λ=245nm) and radioactivity detector;

[0162] Analytical column: GL Sciences, WondaSil C18-WR, 4.6 × 150 mm;

[0163] Column temperature: 20°C;

[0164] Mobile phase solution: CH3CN / H2O=70 / 30;

[0165] Flow rate: 1 mL / min;

[0166] Test results such as Figure 6 , Figure 6 The retention time between the radioactive product and the standard compound was 6.5 min.

[0167] 4. Preparation 18 F-d2-LW223

[0168] 4.1 Automated labeling of drugs using All in One equipment

[0169] (1) Fluorine-18 anion drying method

[0170] Fluorine-18 anions are generated by bombarding H2 with a proton beam in a medical cyclotron. 18 O is produced and transmitted to the All in One device through a pipeline. Waters Sep-Paklight QMA solid phase extraction columns (Tianjin Dexiang Maolong Technology Co., Ltd., 186004051, SEP-PAK LIGHT QMA50BX) were activated with sodium bicarbonate aqueous solution (7.3%) and sterile water for injection, respectively, and then used to capture and separate H2 18 Fluorine-18 anions in O. 1 mL of TEAB eluent (6 mg TEAB in 0.7 mL ACN and 0.3 mL H2O) was automatically drawn with a 3 mL syringe to elute the solid phase extraction column, and the eluent entered the reaction bottle.

[0171] The reaction flask was heated to 85°C and blown with dry nitrogen gas for 5 min. Then 1 mL of acetonitrile was added to the reaction flask and the mixture was evaporated to dryness at 85°C for 5 min and 110°C for 2 min. 18 F-TEAF / TEAB mixture. The reaction flask was then cooled to 40°C.

[0172] (2) Fluorine-18 anion and 18 F-d2-LW223 precursor reaction

[0173] 4.0mg18 The F-d2-LW223 precursor (dissolved in 1.0 mL ACN) was added to the reaction bottle, and the reaction bottle was heated to 110°C for 10 min. The reaction bottle was then cooled to 40°C, and 1.5 mL ACN + 1.0 mL H2O were added to quench the reaction.

[0174] (3) HPLC separation and purification of radioactive products

[0175] 3.5 mL of solution was drawn from the reaction bottle and injected into the loop. In order to reduce the loss of the product in the reaction bottle, 3 mL of acetonitrile was drawn and injected into the reaction bottle again, and then sucked out and injected into the loop again.

[0176] The parameters of HPLC separation and purification are as follows:

[0177] Instrument: Shimadzu HPLC LC-16P;

[0178] Detector: Japan Shimadzu UV detector (SPD-16, λ=254nm) and radioactivity detector;

[0179] Semi-preparative column: OSAKA SODA CAPCELL PAK C18, 5 μm, 10 mm ID × 250 mm;

[0180] Column temperature: 20°C;

[0181] Mobile phase solution: CH3CN / H2O=70 / 30;

[0182] Flow rate: 3 ml per minute;

[0183] Test results such as Figure 7 , Figure 7 The retention time of the product is 17 min.

[0184] 4.2 Automated labeling of drugs using the GE Tracerlab FX2N device

[0185] (1) Fluorine-18 anion drying method

[0186] Fluorine-18 anions are generated by bombarding H2 with a proton beam in a medical cyclotron. 18 O was produced and transferred to the GE Tracerlab FX2N device through a pipeline. A Waters Sep-Pak light QMA solid phase extraction column (Tianjin Dexiang Maolong Technology Co., Ltd., 186004051, SEP-PAK LIGHTQMA 50BX) was activated with sodium bicarbonate aqueous solution (7.3%) and sterile water for injection, respectively, and then used to capture and separate H2 18Fluorine-18 anion in O. Elution with alkaline solution in V6 (6 mg TEAB + 0.7 mL anhydrous ethanol + 0.3 mL sterile water) 18 F - To reaction bottle No. 2, evaporate to dryness at 65°C for 3 min, add the liquid in V7 (1 mL acetonitrile) to reaction bottle No. 2, evaporate to dryness at 65°C and 95°C for 10 min. Then cool the reaction bottle to 50°C.

[0187] (2) Fluorine-18 anion and 18 F-d2-LW223 precursor reaction

[0188] 4.0mg 18 The F-d2-LW223 precursor was dissolved in 1.0 mL ACN and placed in V8, added to the No. 2 reaction bottle, and heated to 110°C for 10 min. Then the No. 2 reaction bottle was cooled to 40°C, and V9 (1.5 mL acetonitrile + 1 mL water) was added to the reaction bottle 2 and transferred to Tube 2 to quench the reaction.

[0189] (3) HPLC separation and purification of radioactive products

[0190] Draw 3.5 mL of product from Tube 2 and insert it into the loop.

[0191] The parameters of HPLC separation and purification are as follows:

[0192] Instrument: Shimadzu HPLC LC-16P

[0193] Detector: Japan Shimadzu ultraviolet detector (SPD-16, λ=245nm) and radioactivity detector were used for joint detection.

[0194] Semi-preparative column: OSAKASODA CAPCELL PAK C18, 5μm, 10mm ID×250mm

[0195] Column temperature: 20℃

[0196] Mobile phase solution: CH3CN / H2O=70 / 30

[0197] Flow rate: 3 ml per minute

[0198] Test results such as Figure 8 , Figure 8 The product retention time is 16 min.

[0199] 4.3 Radioactive product purity detection

[0200] The parameters for purity testing are as follows:

[0201] Instrument: Shimadzu HPLC LC-16P;

[0202] Detector: Japan Shimadzu UV detector (SPD-16, λ=245nm) and radioactivity detector;

[0203] Analytical column: GL Sciences, WondaSil C18-WR, 4.6 × 150 mm;

[0204] Column temperature: 20°C;

[0205] Mobile phase solution: CH3CN / H2O=70 / 30;

[0206] Flow rate: 1 mL / min;

[0207] Test results such as Fig. 9 , Fig. 9 The retention time of the radioactive product and the standard compound was 7.6 min.

[0208] 4.4 Product consistency testing

[0209] Utilization of radioactive products 18 F-d2-LW223 and the control compound d2-LW223 were co-injected and tested, and the test parameters were as follows:

[0210] Instrument: Shimadzu HPLC LC-16P;

[0211] Detector: Japan Shimadzu UV detector (SPD-16, λ=245nm) and radioactivity detector;

[0212] Analytical column: GL Sciences, WondaSil C18-WR, 4.6 × 150 mm;

[0213] Column temperature: 20°C;

[0214] Mobile phase solution: CH3CN / H2O=70 / 30;

[0215] Flow rate: 1 mL / min;

[0216] Test results such as Fig.10 , Fig.10 The retention time of the radioactive product and the standard compound was 7.6 min.

[0217] 2. In vivo activity studies of compounds

[0218] 1. 18 F-d2-LW223, 18 F-d1-LW223 and 18 Comparison of imaging efficacy of F-LW223 in cynomolgus monkeys

[0219] 60 minutes after the drug was injected into the cynomolgus monkey, arterial blood was collected from the cynomolgus monkey and centrifuged to obtain plasma. The radioactive components in the plasma were analyzed to obtain 18 F-d2-LW223 has the best metabolic stability. The SUV normalized by the aorta uptake value was compared, with lumbar vertebrae 3 to 5 as the region of interest. 18 The bone uptake of F-d2-LW223 was lower than 18 F-d1-LW223, 18 F-LW223 (L3: 3.12VS 3.29, P=0.0299; L4: 3.29VS 4.18, P=0.0381; L5: 3.45VS 4.21, P=0.0362). Fig.11 yes 18 F-d2-LW223, 18 F-d1-LW223 and 18 Comparative results of F-LW223 metabolism in cynomolgus monkeys, Fig.11 Medium A is 60 minutes 18 F-d2-LW223, 18 F-d1-LW223 and 18 Results of plasma metabolism analysis of F-LW223 in cynomolgus monkeys. Fig.11 B is 18 F-d2-LW223, 18 F-d1-LW223 and 18 Whole-body static imaging results of F-LW223 in cynomolgus monkeys. Fig.11 The results showed that the deuterated radiopharmaceutical had better plasma metabolic stability and could significantly improve the defluorination of traditional drugs.

[0220] Table 1 is 18 F-d2-LW223 and 18 The statistical results of the pharmacokinetic parameters of F-d1-LW223 are shown in Table 1. 18 V calculated by various pharmacokinetic models for F-d2-LW223 T The values ​​are lower than 18 F-LW223, Description 18 F-d2-LW223 has lower background uptake in brain imaging.

[0221] Table 1 18 F-d2-LW223 and 18 Comparison of pharmacokinetic parameters of F-LW223

[0222]

[0223] 2. 18 Study on imaging of F-d2-LW223 in stroke rat model

[0224] 18 F-d2-LW223 brain imaging in stroke rats showed that PET uptake in the ischemic area was significantly increased compared with the contralateral normal area ( Fig.12 In A and C, after using PK11195, a TSPO-specific blocking agent, the uptake in the lesion area that had previously increased uptake was significantly suppressed, reflecting 18 The specificity of F-d2-LW223 on TSPO expression in the brain ( Fig.12 B and C). Immunofluorescence results in the brain of stroke rats showed increased TSPO expression and microglial activation in the lesion site, which was consistent with the PET imaging results ( Fig.12 (middle D).

[0225] 3. 18 Clinical application study of F-d2-LW223 in stroke patients

[0226] Edaravone and dextroborneol are a compound preparation that uses two ingredients to scavenge free radicals, fight inflammation, and protect the blood-brain barrier through multiple mechanisms, which can significantly improve neuroinflammation caused by acute ischemic stroke. 18 F-d2-LW223 tracer observes the activation state of microglia to verify the neuroprotective mechanism of edaravone and dexamethasone. The clinical study has achieved phased research results. The results show that 18 F-d2-LW223 can well visualize neuroinflammatory changes in the brain ( Fig.13 A), and patients treated with edaravone and dexamethasone, 18 The F-d2-LW223 signal changes showed a slight decrease or remained basically unchanged, while the control group patients 18 F-d2-LW223 uptake increased significantly with time ( Fig.13 The research has achieved preliminary results. 18 F-d2-LW223 PET / MR examination can track and display the microglial activation process with high sensitivity and specificity, reflecting the progression of inflammation.

[0227] 4. 18 Application of F-d2-LW223 in Huntington's disease model

[0228] Huntington's disease is an autosomal dominant, chronic progressive neurodegenerative disease in which neuroinflammation plays an important role in the course of the disease. 18F-d2-LW223 PET imaging can detect a significant increase in whole brain uptake in the disease group ( Fig.14 The important advantage of PET is that it can realize the time longitudinal study of the changes of a certain target in the same group of research subjects. Our team conducted a longitudinal study of the Huntington's disease model at multiple time periods (1.5 months, 3 months, 7 months, 10 months, and 13 months of age). 18 F-d2-LW223 PET imaging showed that 18 F-d2-LW223 uptake increases with age ( Fig.15 Immunofluorescence experiments confirmed the imaging effect of TSPO ( Fig.16 ). In summary, 18 F-d2-LW223PET can track the continued activation of microglia during the chronic course of Huntington's disease and further confirm 18 Imaging efficacy of F-d2-LW223 PET in neuroinflammation.

[0229] All documents mentioned in this application are cited as references in this application, just as each document is cited as reference separately. Unless they conflict with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited with all contents and all purposes. When the cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When the cited documents are involved in this application, the examples and preferred methods of the cited relevant technical features can also be incorporated into this application as references, but are limited to the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be modified adaptively according to the description of this application.

[0230] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0231] The above-described embodiments only express several implementation methods of the present application, but they cannot be understood as limiting the scope of the patent application. It should be pointed out that, for those of ordinary skill in the art, without departing from the concept of the present application, several variations and improvements can also be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by those skilled in the art on the basis of the technical solutions provided in the present application through logical analysis, reasoning or limited experiments are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. An organic compound having a structure as shown in formula (I): in, M1 and M2 are each independently selected from D or H, and at least one of M1 and M2 is D; X1 from F 18 or F.

2. The organic compound according to claim 1, characterized in that Both M1 and M2 are D.

3. A method for preparing an organic compound according to claim 1 or 2, characterized in that: The steps include: Compound A is reacted with a deuterated reducing agent to reduce the ester group and deuterate the hydrogen at the ortho position to obtain compound B; The compound B is subjected to oxidation and esterification reaction to obtain compound C; The compound C is ring-opened and subjected to an amidation reaction with the compound D to obtain the compound E; The OH of the compound E is subjected to a halogenation reaction with an X2 donor reagent to obtain a compound F, wherein X2 is selected from Br or Cl; X2 of the compound F is subjected to a halogen exchange reaction with a donor reagent X1 to prepare an organic compound having a structure as shown in formula (I).

4. The preparation method according to claim 3, characterized in that: The deuterated reducing agent comprises at least one of deuterated lithium aluminum hydride and deuterated sodium borohydride; and / or The molar ratio of the compound A to the deuterated reducing agent is 1:(3.5-4.5).

5. The preparation method according to claim 3, characterized in that: The compound B is subjected to oxidation and esterification reaction, comprising: Mix the compound B and the oxidant at 15°C±1°C, and carry out reduction and esterification reaction at 25°C-35°C for 25-35 minutes; Optionally, the oxidant is selected from chromate pyridinium salt oxidants; Optionally, the molar ratio of the compound B to the oxidant is 1:(2-3).

6. The preparation method according to claim 3, characterized in that: The compound C is ring-opened and subjected to an amidation reaction with the compound D, comprising: Add Lewis acid to the organic solution of compound D at 15°C±1°C, and stir at 25°C-35°C for 10-20 minutes to obtain a first mixed solution; Adding the compound C to the first mixed solution, and subjecting to thermal reflux reaction for 24 hours to 48 hours; Optionally, the molar ratio of the compound C to the compound D is 1:(4-4.5); Optionally, the molar ratio of the Lewis acid to the compound D is 1:(3-3.2).

7. The preparation method according to claim 3, characterized in that: The temperature of the halogen exchange reaction is 110°C ± 5°C, and the time is 8 min to 12 min.

8. A PET probe for an 18 kDa translocator protein, characterized in that: The organic compound comprises any one of claims 1 to 7.

9. Use of the organic compound according to any one of claims 1 to 7 in the preparation of a medicament for treating neuritis.

10. The use according to claim 9, characterized in that: The organic compound treats neuritis by targeting the 18 kDa translocator protein.

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