Probe for MAGL

Abstract

The present invention provides compounds having the general formula (I) wherein R1, R2, X and Y are as described herein; compositions comprising the compounds; methods of making the compounds; and methods of using the compounds.

Description

Technical Field

[0001] This invention relates to organic compounds that can be used as fluorescent, Raman, or PET probes for monoacylglycerol lipase (MAGL). Background Technology

[0002] Fluorescent imaging probes have become high-resolution tools for studying localization, such as expression levels and protein distribution in healthy and disease states, and the structure, dynamics, and function of proteins in living cells (LA Stoddart, LE Kilpatrick, SJ Briddon, SJ Hill, Neuropharmacology 2015, 98, 48–57). These probes can be used, for example, with confocal live-cell imaging in flow cytometry fluorescence activated cell sorting (FACS) experiments or cell transport studies. Furthermore, fluorescent imaging probes allow for real-time monitoring of ligand-receptor interactions and protein visualization with high spatiotemporal precision (AJ Vernall, SJ Hill, B. Kellam, Br. J. Pharmacol. 2014, 171, 1073–1084; C. Iliopoulos-Tsoutsouvas, RN Kulkarni, A. Makriyannis, SP Nikas, Expert Opin. Drug Discov. 2018, 13, 933–947). Furthermore, these probes offer the potential to generate equilibrium and kinetic binding data in a high-throughput manner without the need for treatment of radioactive materials, such as time-resolved fluorescence resonance energy transfer (TR-FRET). Fluorescent imaging probes can also be used to support the translation of preclinical pharmacology animal data into clinical pharmacology animal data, and can be applied to dose selection in humans. They can be used, for example, as biomarkers for target binding by generating ex vivo quantitative receptor binding data in whole blood. Depending on their respective applications, fluorescent imaging probes need to be matched to specific criteria, including affinity for their respective targets, selectivity and specificity, favorable photophysical properties, and suitability across different technologies and cell types.

[0003] Similarly, Raman spectroscopy probes can monitor cellular and tissue changes in real time, which is helpful for studying drug-target binding (K. Antonio, Z. Schultz Anal. Chem., 2014, 86(1) 30-46; C. Krafft, J. PoppAnal. Bioanal. Chem., 2015, 407(3) 699-717; Lawson et al., J. Raman Spectrosc., 1997, 28(2–3), 111–117). Compared with fluorescent probes, only the specific Raman absorption of the probe, rather than its fluorescence properties, is relevant. The absorption range for reliable Raman spectroscopy analysis of biological samples is typically in the 1800 cm⁻¹ range. - ¹and 2800 cm - Between ¹, the Raman signal is easy to detect and distinguish.

[0004] [ 18 [F] Positron emission tomography (PET) probes labeled with the radioactive isotope fluorine-18 have significant advantages over fluorescent probes in biomedical imaging (S. Chua, A. Groves, Biomedical Imaging: Applications and Advances, 2014). First, [ 18 F-PET probes, due to their deep tissue penetration and high spatial resolution, allow for precise localization of molecular targets within organisms. Secondly, their radioactive properties enable accurate quantitative assessment of physiological processes and molecular interactions within the body. Fluorescent probes and [ 18 The advantages of each of the F-PET probes can be seen through bimodal [ 18 F]-BODIPY probe combo (Y. Kwon et al., Nucl. Med. Biol. 2021, 93, 22-36). Summary of the Invention

[0005] In a first aspect, the present invention provides a compound of formula (I). (I) Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 X and Y are as defined in this article.

[0006] In another aspect, the present invention provides compositions comprising formula (I), methods for manufacturing formula (I), and methods for using formula (I). Attached Figure Description

[0007] Figure 1 illustrates the process described in Example 2. 18 HPLC chromatogram of F-labeled compounds. Detailed Implementation

[0008] definition

[0009] Features, integrals, properties, compounds, chemical parts, or groups described in connection with a particular aspect, embodiment, or example of the invention should be understood to be applicable to any other aspect, embodiment, or example described herein, unless incompatible therewith. All features disclosed in this specification (including any appended claims, abstract, and drawings) and / or all steps of any method or process disclosed thereby may be combined in any combination, except for at least some mutually exclusive combinations of such features and / or steps. The invention is not limited to the details of any of the foregoing embodiments. The invention extends to any novel feature or any novel combination of features disclosed in this specification (including any appended claims, abstract, and drawings), or to any novel step or any novel combination of steps of any method or process disclosed thereby.

[0010] The term "alkyl" refers to a monovalent or polyvalent (e.g., monovalent or divalent) straight-chain or branched saturated hydrocarbon group ("C1-C6-alkyl") having 1 to 6 carbon atoms (e.g., 1, 2, 3, 4, 5, or 6 carbon atoms). In other embodiments, the alkyl group contains 1 to 3 carbon atoms, such as 1, 2, or 3 carbon atoms. Some non-limiting examples of alkyl groups include methyl, ethyl, propyl, 2-propyl (isopropyl), n-butyl, isobutyl, sec-butyl, tert-butyl, and 2,2-dimethylpropyl. A particularly preferred but non-limiting example of an alkyl group is methyl.

[0011] The term "haloalkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group has been replaced by a halogen atom, preferably fluorine. Preferably, "haloalkyl" refers to an alkyl group in which one, two, or three hydrogen atoms of the alkyl group have been replaced by a halogen atom, most preferably fluorine. Particularly preferred but non-limiting examples of haloalkyl are trifluoromethyl (CF3) and 2,2,2-trifluoroethyl (CF3CH2-).

[0012] The term "hydroxyalkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group has been replaced by a hydroxyl group. Preferably, "hydroxyalkyl" refers to an alkyl group in which one, two, or three hydrogen atoms of the alkyl group have been replaced by a hydroxyl group. A particularly preferred but non-limiting example of a hydroxyalkyl group is hydroxymethyl.

[0013] The term "halogen" or "halogenated" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). Preferably, the term "halogen" or "halogenated" refers to fluorine (F), chlorine (Cl), or bromine (Br). Particularly preferred, but non-limiting, examples of "halogen" or "halogenated" are fluorine (F) and chlorine (Cl).

[0014] The term "heteroaryl" refers to an aromatic monovalent or polyvalent monocyclic ring system containing one or more heteroatoms and having a total of 5 to 6 ring members. Preferably, "heteroaryl" refers to a 5- to 6-membered heteroaryl containing 1, 2, 3, or 4 heteroatoms independently selected from O, S, and N. More preferably, "heteroaryl" refers to a 5- to 6-membered heteroaryl containing 1 to 3 heteroatoms independently selected from O, S, and N. Most preferably, "heteroaryl" refers to a 5-membered heteroaryl containing 1 to 2 heteroatoms independently selected from O, S, and N. Preferred but non-limiting examples of heteroaryl include pyrroleyl.

[0015] The term "pharmaceutically acceptable salt" refers to those salts that retain the biological effects and properties of a free base or free acid, and are not undesirable in biological or other respects. These salts are formed using inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. (especially hydrochloric acid) and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethylsulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, etc. Furthermore, these salts can be prepared by adding an inorganic or organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of the following: primary, secondary, and tertiary amines; substituted amines (including naturally occurring substituted amines); cyclic amines; and basic ion exchange resins (such as isopropylamine, trimethylamine, diethylamine, hydroxylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyimide resins, etc.). Specific pharmaceutically acceptable salts of compounds of formula (I) are hydrochloride salts.

[0016] Compounds of formula (I) may contain several asymmetric centers and may exist as optically pure enantiomers, mixtures of enantiomers (e.g., racemates), optically pure diastereomers, mixtures of diastereomers, diastereomers, or mixtures of diastereomers.

[0017] According to the Cahn-Ingold-Prelog specification, asymmetric carbon atoms can have either an "R" or an "S" configuration.

[0018] The abbreviation "MAGL" refers to monoacylglycerol lipase. The terms "MAGL" and "monoacylglycerol lipase" are used interchangeably in this document.

[0019] The compounds of the present invention

[0020] In a first aspect, the present invention provides a compound of formula (I). (I) Or its pharmaceutically acceptable salt, wherein: X is CH and Y is CH2; or X and Y together form the group CH=C; R 1 Selected from: ; R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; and R 2 Selected from: , and ; R 3a and R 3b Each is independently selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; The reporting unit is selected from: ; ;and ; L is selected from –(CH2) p –, –(CH2)2-NHC(O)-CH2–, –CH2-NHC(O)-(CH2)2–, –((CH2)2O) q -(CH2)2– and –(CH2)2-(O(CH2)2) q –; n and m are each independently 1 or 2; p is selected from 1, 2, 3, 4, and 5; q is selected from 1, 2, and 3; and r is selected from 1, 2, 3, and 4.

[0021] In a first aspect, the present invention provides a compound of formula (I). (I) Or its pharmaceutically acceptable salt, wherein: X is CH and Y is CH2; or X and Y together form the group CH=C; R 1 Selected from: ; R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; and R 2 Selected from: , and ; R 3a and R 3b Each is independently selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; The reporting unit is selected from: ; ;and ; L is selected from –(CH2) p –, –(CH2)2-NHC(O)-CH2–, –CH2-NHC(O)-(CH2)2–, –((CH2)2O) q -(CH2)2– and –(CH2)2-(O(CH2)2) q –; n and m are each independently 1 or 2; p is selected from 1, 2, 3, 4, and 5; and q is selected from 1, 2, and 3.

[0022] In a preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein X is CH and Y is CH2.

[0023] In one embodiment, the invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 1 for , where R 1a As described in this article.

[0024] In one embodiment, the invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 1 for , where R 1a It is a halogenated-C1-C6-alkyl group.

[0025] In a preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 1 for , where R 1a Selected from CF3 and hydroxymethyl.

[0026] In a particularly preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 1 for , where R 1a It is CF3.

[0027] In one embodiment, the invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , and ; Where R 3a R 3b R 3c m, n, and r are as defined in this article.

[0028] In one embodiment, the invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , and ; Where R 3a R 3b R 3c m and n are as defined in this article.

[0029] In one embodiment, the invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , and ;in: R 3a Selected from hydrogen and C1-C6-alkyl; R 3b Selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; m is 1 or 2; and n is 1.

[0030] In a preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , and ;in: R 3a Selected from hydrogen and methyl; R 3b Selected from hydrogen, methyl, and pyrrole groups; R 3c Selected from F and 18 F; m is 1 or 2 n is 1.

[0031] In another preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , , , and ;in: R 3c Selected from F and 18 F.

[0032] In a particularly preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , and ;in: R 3c Selected from F and 18 F.

[0033] In a preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein R 3c It is F.

[0034] In one embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein: X is CH and Y is CH2; X and Y together form the group CH=C; R 1 for ; R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; R 2 Selected from: , and ;in: R 3a Selected from hydrogen and C1-C6-alkyl; R 3b Selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; m is 1 or 2; and n is 1.

[0035] In a preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein: X is CH and Y is CH2; R 1 for ; R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; R 2 Selected from: , , , , , and ;in: R 3c Selected from F and 18 F.

[0036] In a particularly preferred embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein: X is CH and Y is CH2; R 1 for ; R 1a Selected from CF3 and hydroxymethyl; R 2 Selected from: , , , , , and ;in: R 3c It is F.

[0037] In one embodiment, the present invention provides a compound of formula (I) as described herein, or a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) is selected from: 6-((5,5-difluoro-5H-4λ4,5λ4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid[ 18 F]-1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3-methyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3,7-dimethyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester; 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester; 6-(2-(2-(3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-3-yl)propamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-(2-(2-(4-((4-(dimethylamino)phenyl)but-1,3-diyne-1-yl)benzamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; and N-(10-(2-carboxy-5-((5-(2-((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-yl)methyl)phenoxy)pentyl)carbamoyl)phenyl)-7-(dimethylamino)-5,5-dimethyldibenzo[b,e]silicyclohexanetriene-3(5H)-ylidene)-N-methylmethylammonium.

[0038] In one particular embodiment, the present invention provides a pharmaceutically acceptable salt of a compound of formula (I) as described herein. In another particular embodiment, the present invention provides a compound of formula (I) as described herein as a free base.

[0039] In some embodiments, compounds of formula (I) are isotopically labeled by replacing one or more of their atoms with atoms having different atomic masses or mass numbers. Such isotopically labeled (i.e., radioactively labeled) compounds of formula (I) are considered to be within the scope of this disclosure. Examples of isotopes that can be incorporated into compounds of formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as, but not limited to, isotopes of these elements. 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 36 Cl、 123 I and 125 I. Certain isotope-labeled compounds of formula (I) (e.g., those containing a radioactive isotope) can be used for drug and / or matrix tissue distribution studies. Radioactive isotope tritium (i.e....) 3 H) and carbon-14 (i.e. 14 C) This is particularly useful because they are easy to incorporate and detection methods are readily available. For example, compounds of formula (I) can be enriched with a given isotope of 1%, 2%, 5%, 10%, 25%, 50%, 75%, 90%, 95%, or 99%.

[0040] Using heavier isotopes (such as deuterium, i.e.) 2 H) Substitution can provide certain therapeutic advantages due to greater metabolic stability, such as increased in vivo half-life or reduced dose requirements.

[0041] Using positron emission isotopes (such as...) 11 C 18 F, 15 O and 13 N) can be used to examine substrate acceptor occupancy in positron emission tomography (PET) studies. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the examples set forth below, using an appropriate isotopically labeled reagent instead of the previously used unlabeled reagent.

[0042] Preparation process

[0043] The preparation of compounds of formula (I) of the present invention can be carried out via sequential or concurrent synthetic routes. The synthesis of the present invention is illustrated in the following general scheme. The skills required to perform the reactions and purify the resulting products are known to those skilled in the art. Unless otherwise specified, the substituents and indices used in the following description of the methods have the meanings provided herein.

[0044] If any of the starting material, intermediate, or compound of formula (I) contains one or more functional groups that are unstable or reactive under the reaction conditions of one or more reaction steps, a suitable protecting group may be introduced prior to the critical steps of a process well known in the art (as described, for example, in TW Greene and PGM Wutts, “Protective Groups in Organic Chemistry,” 5th edition, 2014, John Wiley & Sons, NY). Such protecting groups may be removed later in the synthesis using standard methods described in the literature.

[0045] If the starting material or intermediate contains a stereoisomeric center, the compound of formula (I) can be obtained in the form of diastereomers or mixtures of enantiomers, which can be separated by methods well known in the art, such as chiral HPLC, chiral SFC, or chiral crystallization. Racemic compounds can be separated into their corresponding counterparts, for example, by diastereomeric salts, which are separated by crystallization with optically pure acids, or by specific chromatographic methods using chiral adsorbents or chiral eluents. Similarly, starting materials and intermediates containing stereoisomeric centers can be separated to provide diastereomer / enantiomer-enriched starting materials and intermediates. The use of such diastereomer / enantiomer-enriched starting materials and intermediates in the synthesis of compounds of formula (I) will generally yield diastereomer / enantiomer-enriched compounds of the corresponding formula (I).

[0046] Those skilled in the art will recognize that, if not desired, an "orthogonal protecting group strategy" will be applied in the synthesis of compounds of formula (I), which allows multiple protecting groups to be cleaved at a time without affecting other protecting groups in the molecule. The principle of orthogonal protection is well known in the art and has been reported in the literature (e.g., Barany and R.B. Merrifield, J. Am. Chem. Soc. 1977, 99, 7363; H. Waldmann et al., Angew. Chem. Int. Ed. Engl. 1996, 35, 2056).

[0047] Those skilled in the art will recognize that the reaction sequence can vary depending on the reactivity and properties of the intermediate.

[0048] More specifically, the compound of formula (I) can be prepared by the methods given below, by the methods given in the examples, or by similar methods. Appropriate reaction conditions for each reaction step are known to those skilled in the art. Likewise, for information on reaction conditions affecting the reactions reported in the literature, see, for example: *Comprehensive Organic Transformations: A Guide to Functional Group Preparations*, 2nd edition, Richard C. Larock, John Wiley & Sons, New York, NY. 1999). The reaction can be carried out with or without a solvent. There are no particular limitations on the nature of the solvent used, as long as it does not adversely affect the reaction or the reagents involved and is at least partially capable of dissolving the reagents. The described reaction can occur over a wide temperature range, and precise reaction temperatures are not critical to the invention. The above reactions can be conveniently carried out in the temperature range from -78°C to reflux. The reaction time can also vary considerably depending on many factors, particularly the reaction temperature and the nature of the reagents. However, it typically takes from 0.5 hours to several days to obtain the intermediates and compounds described. The reaction sequence is not limited to the order shown in the scheme; however, the order of reaction steps can be freely changed depending on the starting materials and their corresponding reactivity.

[0049] If the starting materials or intermediates are not commercially available, or their synthesis is not described in the literature, they can be prepared in a manner similar to existing procedures for closely analogous products or as outlined in the experimental section.

[0050] Compounds of Formula I can be processed according to procedures similar to those in the literature and / or as follows: Option 1 The synthesis described in the text.

[0051]

[0052] Option 1

[0053] The commercially available starting material 6-oxo-2-azaspiro[3.3]heptane-2-carboxylate tert-butyl ester (1) can be converted to 6-((4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylate tert-butyl ester (2) via a so-called boron-Wittig olefination reaction, as described in the literature (Cuenca and Fernández 2021; Kovalenko et al. 2019). The resulting boric acid compound is a general chemical structural unit that can be used in various cross-coupling reactions (Suzuki, Liebeskind-Srogl, Chan-Lam, etc.). By the Suzuki reaction, it can be coupled with various substituted aromatic moieties with halogen (Cl, Br, I) or pseudohalogen (including trifluoromethanesulfonate, methanesulfonate, etc.) leaving groups to give intermediates of general formula 3. As those skilled in the art will know, these reactions are typically carried out under palladium catalysis using various phosphine ligands (i.e., PPh3). The reactions are conducted in suitable solvents such as water, 1,4-dioxane, acetonitrile, THF, or mixtures thereof, at suitable reaction temperatures between 0°C and reflux temperatures, with typical reaction times ranging from 30 minutes to 24 hours. The N-boc-protected intermediate 3 can be readily deprotected under a wide range of acidic or basic conditions, most commonly with mixtures of TFA and DCM in varying proportions. Deprotected secondary amines can be coupled with R under alkaline conditions (TEA, DIPEA, NaHCO3, K2CO3, NaH, etc.) in appropriate solvents (DCM, MeCN, DMF, etc.) at temperatures between 0°C and reflux temperature, using various coupling reagents (CDI, CDT, triphosgene, bis-(pentafluorophenyl)-carbonate, etc.), or directly via their corresponding activated analogues (CDT, 4-nitrobenzene chloroformate, bis-(hexafluoroisopropyl)carbonate, or N,N′-disuccinimidyl carbonate) and R. 1The nucleophilic group is coupled to give intermediate 4. Those skilled in the art will recognize that the now-activated carbamates (4 to 11) should be handled with extreme care as they readily react with nucleophiles, including water, especially under basic conditions. In the hydrogenation step, intermediate 4 can be converted to 5, wherein two hydrogenation steps are carried out in one pot: hydrogenation of the CC double bond and the primary amine Cbz protecting group. This reaction can be carried out with elemental hydrogen or a hydrogen-generating substance (i.e., silane) in the presence of a suitable catalyst (most commonly palladium on activated carbon). Those skilled in the art will recognize that the Cbz protection strategy is an effective tool for obtaining the desired intermediate, but other protecting groups may be applicable (compare, e.g., Greene and Wuts, Protective Groups in Organic Synthesis, 1999). In the final step of obtaining a covalent probe having the general formula 6 as defined in claim 1, the corresponding reporter unit must be coupled to the corresponding carboxylic acid or acyl chloride via an amide formation reaction. The reaction is typically carried out via coupling reagents (i.e., HATU, HBTU, HOBt, DCC, etc.), under base catalysis (i.e., TEA, DIPEA, DBU, K2CO3, etc.), in a suitable solvent (DMF, DMA, MeCN, THF, DMSO, etc.), at a temperature between 0°C and reflux temperature.

[0054] Intermediate 2 can also be converted into intermediate 7, in the same manner as the conversions from 3 to 4 described above. Boric acid intermediate 7 can be hydrolyzed to the corresponding boric acid 8, typically using a buffer such as ammonium acetate and an oxidizing agent such as NaIO4 in a suitable co-solvent such as acetone. The corresponding boric acid can then be coupled via Liebeskind-Srogl cross-coupling (LSCC) to an 8-thiomethyl-substituted BODIPY analog, as described in the literature (Arroyo et al. 2011; de J. Gómez-Infante et al. 2016) to obtain intermediate 9. The CC double bond can be hydrogenated as described above to obtain the fluorescent covalent probe 10 as claimed in claim 1.

[0055] Bimodal fluorescent and radiolabeled compounds of general formula 11 can be obtained through radiolabeling techniques, one of which 19 F atoms are radioactive 18 F substitution, as described in the literature for these BODIPY structures (Hendricks et al. 2012; Keliher et al. 2014; Kwon et al. 2021).

[0056] References: Antonio, Karen A., and Zachary D. Schultz. 2014. “Advances in Biomedical Raman Microscopy.” Analytical Chemistry 86(1): 30–46. doi: 10.1021 / ac403640f. Arroyo, Ismael J., Rongrong Hu, Ben Zhong Tang, Fabiola I. López, and Eduardo Peña-Cabrera. 2011. “8-Alkenylborondipyrromethene Dyes. General Synthesis, Optical Properties, and Preliminary Study of Their Reactivity.” Tetrahedron 67(38): 7244–50. doi: 10.1016 / j.tet.2011.07.067. Chua, S., and A. Groves. 2014. Biomedical Positron Emission Tomography (PET) Imaging. Cisar, Justin S., Olivia D. Weber, Jason R. Clapper, Jacqueline L. Blankman, Cassandra L. Henry, Gabriel M. Simon, Jessica P. Alexander, Todd K. Jones, R. Alan B. Ezekowitz, Gary P. O’Neill, and Cheryl A. Grice. 2018. “Identification of ABX-1431, a Selective Inhibitor of Monoacylglycerol Lipase and Clinical Candidate for Treatment of Neurological Disorders.” Journal of Medicinal Chemistry 61(20): 9062–84. doi: 10.1021 / acs.jmedchem.8b00951. Cuenca, Hannah B., and Elena Fernandez. “Boron-White Olefination with Gem -Bis(Boryl)Alkanes.” . Chemical Society Reviews 50(1):72–86. doi:10.1039 / D0CS00953A. Hendricks, J. Adam, Edmund J. Keliher, Dongpeng Wan, Scott A.Hilderbrand, Ralph Weissleder, and Ralph Mazitschek.2012. “Synthesis of [ 18F]BODIPY: A Bifunctional Reporter for Hybrid Optical / Positron Emission TomographyImaging.” Applied Chemistry - International Edition 51(19):4603–6. doi:10.1002 / anie.201107957. of J. Gomez-Infante, Antonio, Jorge Bañuelos, Ismael Valois-Escamilla, David Cruz-Cruz, Ruth Prieto-Montero, Inigo Lopez-Arbeloa, TeresaArbeloa, and Eduardo Peña-Cabrera. “Synthesis, Properties, andFunctionalization of Nonsymmetric 8-MethylthioBODIPYs.” European Journal ofOrganic Chemistry 2016(29):5009–23. doi: 10.1002 / ejoc.201600724. Keliher, Edmund J., Jenna A. Klubnick, Thomas Reiner, Ralph Mazitschek, and Ralph Weissleder. 2014. “Efficient Acid-Catalyzed 18F / 19F Fluoride Exchange of BODIPY Dyes.” ChemMedChem 9(7): 1368–73. doi: 10.1002 / cmdc.201300506. Kovalenko, Maksym, Dmytro V. Yarmoliuk, Dmytro Serhiichuk, Daria Chernenko, Vladyslav Smyrnov, Artur Breslavskyi, Oleksandr V. Hryshchuk, Ihor Kleban, Yuliya Rassukana, Andriy V. Tymtsunik, Andrey A. Tolmachev, Yuliya O. Kuchkovska, and Oleksandr O. Grygorenko. 2019. “The Boron-Wittig Olefination of Aldehydes and Ketones with Bis[(Pinacolato)Boryl]Methane: An Extended Reaction Scope.” European Journal of Organic Chemistry 2019(33): 5624–35. doi: 10.1002 / ejoc.201900648. Krafft, Christoph, and Jürgen Popp. 2015. “The Many Facets of Raman Spectroscopy for Biomedical Analysis.” Analytical and Bioanalytical Chemistry 407(3): 699–717. doi: 10.1007 / s00216-014-8311-9. Kwon, Young Do, Youngjoo Byun, and Hee Kwon Kim. 2021. “18F-Labelled BODIPY Dye as a Dual Imaging Agent: Radiofluorination and Applications in PET and Optical Imaging.” Nuclear Medicine and Biology 93:22–36. doi: 10.1016 / j.nucmedbio.2020.11.004. Lawson, E. E., B. W. Barry, A. C. Williams, and H. G. M. Edwards. 1997. “Biomedical Applications of Raman Spectroscopy.” Journal of Raman Spectroscopy 28(2–3):111–17. doi: 10.1002 / (sici)1097-4555(199702)28:2 / 3<111::aid-jrs87>3.0.co;2-z. Schaus, Scott E., Bridget D. Brandes, Jay F. Larrow, Makoto Tokunaga, Karl B. Hansen, Alexandra E. Gould, Michael E. Furrow, and Eric N. Jacobsen. 2002. “Highly Selective Hydrolytic Kinetic Resolution of Terminal Epoxides Catalyzed by Chiral (Salen)CoIII Complexes. Practical Synthesis of Enantioenriched Terminal Epoxides and 1,2-Diols.” Journal of the American Chemical Society 124(7):1307–15. doi: 10.1021 / ja016737l.

[0057] MAGL inhibitory activity

[0058] The enzymatic activity of the compound against MAGL was determined by hydrolyzing the natural substrate 2-arachidonicylglycerol (2-AG) to obtain arachidonic acid, followed by analysis of the compound's inhibitory activity against MAGL, and then mass spectrometry analysis. This determination is referred to below as the "2-AG determination".

[0059] 2-AG assays were performed in 384-well plates (PP, Greiner catalog number 784201) with a total volume of 20 µL. Compound dilutions were prepared in 100% DMSO (VWR Chemicals 23500.297) using a 3-fold dilution step to achieve a final concentration range of 12.5 µM to 0.8 pM. 0.25 µL of the compound dilution (100% DMSO) was added to 9 µL of MAGL in the assay buffer (50 mM TRIS (GIBCO, 15567-027), 1 mM EDTA (Fluka, 03690-100 mL), 0.01% (v / v) Tween). After shaking, the plate was incubated at RT for 15 min. 10 µL of 2-arachidonicylglycerol assay buffer was added to initiate the reaction. The final concentrations in this assay were 50 pM MAGL and 8 µM 2-arachidonic acid glycerol. After shaking and incubation at RT for 30 min, 40 µL of ACN containing 4 µM d8-arachidonic acid was added to quench the reaction. The arachidonic acid concentration was monitored using an online SPE system (Agilent Rapidfire) coupled with a triple quadrupole mass spectrometer (Agilent 6460). A C18 SPE column (G9205A) was used in the ACN / water liquid position. The mass spectrometer was operated in negative electrospray mode, with the mass ion pair of arachidonic acid changing from 303.1 to 259.1 and that of d8-arachidonic acid from 311.1 to 267.0. The activity of the compounds was calculated based on the intensity ratio of [arachidonic acid / d8-arachidonic acid].

[0060] Table 1

[0061] In one aspect, the present invention provides compounds of formula (I) as described herein, and pharmaceutically acceptable salts or esters thereof, wherein the compounds of formula (I) and pharmaceutically acceptable salts or esters thereof have an MAGL inhibitory concentration of less than 25 µM, preferably less than 10 µM, more preferably less than 5 µM. 50 This value was measured as described in the MAGL determination described in this article.

[0062] In one embodiment, compounds of formula (I) as described herein and their pharmaceutically acceptable salts or esters have an IC50 between 0.000001 µM and 25 µM. 50 (MAGL inhibition) values, specific compounds have IC50 values ​​between 0.000005 µM and 10 µM. 50 Values, and for other specific compounds, IC50 values ​​are between 0.00005 µM and 5 µM. 50 The value is as measured in the MAGL determination described herein.

[0063] Using the compounds of the present invention

[0064] Compounds of formula (I) are fluorescence imaging, Raman, or PET probes with high affinity for MAGL. Therefore, they can be used as high-resolution tools to study localization, such as expression levels and protein distribution in healthy and disease states, and the structure, dynamics, and function of MAGL in living cells. They can also be applied, for example, using confocal live-cell imaging in flow cytometry fluorescence activated cell sorting (FACS) experiments or cell transport studies.

[0065] In one aspect, the present invention provides a compound of formula (I) as described herein for the study of the occupancy of monoacylglycerol lipase (MAGL).

[0066] In another aspect, the present invention provides compounds of formula (I) as described herein for diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals.

[0067] In another aspect, the present invention provides compounds of formula (I) as described herein for generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

[0068] In another aspect, the present invention provides the use of a compound of formula (I) as described herein in the study of the occupancy of monoacylglycerol lipase (MAGL).

[0069] In another aspect, the present invention provides the use of the compound of formula (I) described herein in diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals.

[0070] In another aspect, the present invention provides the use of compounds of formula (I) as described herein for generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

[0071] In another aspect, the present invention provides a method for studying the occupancy of monoacylglycerol lipase (MAGL), the method comprising contacting MAGL with a compound of formula (I) described herein.

[0072] In another aspect, the present invention provides a method for diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals, the method comprising contacting MAGL with a compound of formula (I) described herein.

[0073] In another aspect, the present invention provides a method for generating equilibrium and kinetic binding data of monoacylglycerol lipase (MAGL), the method comprising contacting MAGL with a compound of formula (I) as described herein.

[0074] Example

[0075] The invention will be more fully understood by referring to the following examples. However, the claims should not be construed as limiting the scope of the examples.

[0076] In the case of preparation examples obtained as mixtures of enantiomers, pure enantiomers can be separated by the methods described herein or by methods known to those skilled in the art, such as chiral chromatography (e.g., chiral SFC) or crystallization.

[0077] Unless otherwise specified, all reaction examples and intermediates are prepared under an argon atmosphere.

[0078] Example 1: 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((5,5-difluoro-5H-4λ4,5λ4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid

[0079] Step 1) 6-((4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester 2,2,6,6-Tetramethylpiperidine (311 mg, 2.2 mmol) was dissolved in anhydrous THF (20 mL) in an oven-dried vial equipped with a magnetic stir bar and cooled to –78 °C under a nitrogen atmosphere. nBuLi (2.5 M in hexane) (880 µL, 2.2 mmol) was added dropwise, and the reaction mixture was stirred at the same temperature for 30 min. Next, a solution of bis(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methane (590 mg, 2.2 mmol) in THF (8 mL) was added dropwise. The reaction was stirred for 5 min. Then, tert-butyl 6-oxo-2-azaspiro[3.3]heptane-2-carboxylate (232 mg, 1.1 mmol) was added dropwise over 5 min in THF (12 mL). The reaction vial was slowly heated to room temperature overnight. After completion, the reaction mixture was exposed to air and filtered through a short silica gel column using diethyl ether as the eluent. The mixture was concentrated under reduced pressure and adsorbed onto the solvent. The product was obtained as a white amorphous solid by silica gel chromatography in 0 to 20% ethyl acetate in cyclohexane. LC-MS (ESI): m / z = 336.1 [M+H] + .

[0080] 1 H NMR (300 MHz, CDCl3) δ 5.18 (p, J = 2.3 Hz, 1H), 4.00 – 3.87 (m,4H), 3.10 – 3.05 (m, 2H), 2.93 (dq, J = 3.3, 1.9 Hz, 2H), 1.43 (s, 9H), 1.23(s, 12H).

[0081] Step 2) 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid: The intermediate was synthesized by first deprotecting 6-((4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (67 mg, 200 µmol) by stirring in 20% TFA in DCM for 3 hours, followed by removal of any volatiles under reduced pressure. The deprotected amine was dissolved in 10 mL of anhydrous DCM and DIPEA (45 µl, 260 µmol) was added. The mixture was cooled to 0 °C and then a solution of bis(1,1,1,3,3,3-hexafluoropropane-2-yl) carbonate ~50 wt% in Et2O (1 equivalent, freshly prepared according to the literature (Cisar et al., 2018)) was added. The mixture was heated to room temperature and stirred for 3 hours. The mixture was diluted with 10 mL of DCM and washed with saturated NaCl solution (5 mL). The organic layer was dried over MgSO4, filtered, and adsorbed onto silica. It was then purified by silica gel chromatography with an elution gradient of 0–20% EtOAc in cyclohexane. The product was obtained as a colorless crystalline solid (67 mg, 78%).

[0082] 1 H NMR (300 MHz, CDCl3) δ 5.64 (hept, J = 6.2 Hz, 1H), 5.22 (p, J =2.3 Hz, 1H), 4.20 – 4.06 (m, 4H), 3.14 (t, J = 2.5 Hz, 2H), 3.00 – 2.97 (m,2H), 1.23 (s, 12H).

[0083] Step 3) ((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid Pinaryl borate (1.0 equivalent), sodium periodate (5.0 equivalent), and ammonium acetate (5.0 equivalent) were dissolved in acetone / water 2:1 (0.05 M boric acid compound) and stirred at ambient temperature for 18 to 24 hours until LC-MS indicated consumption of the starting material. Acetone was removed under reduced pressure, 4 mL of ACN / H₂O 1:1 was added, and the mixture was filtered. The filtrate was purified by RP-HPLC from 5% to 75% ACN:H₂O (+ 0.1% TFA) to give the product as a colorless solid.

[0084] 1H NMR (300 MHz, MeOD) δ 6.02 (hept, J = 6.4 Hz, 1H), 5.34 (p, J =2.3 Hz, 1H), 4.15 (s, 2H), 4.09 (s, 2H), 3.11 (p, J = 2.5 Hz, 2H), 3.01 (d, J= 2.2 Hz, 2H).

[0085] Step 4) 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester ((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid and 8-thiomethyl-BODIPY (1.0 mol) were dissolved in anhydrous THF. The mixture was purged with nitrogen for 10 min. Then copper thiophene-2-carboxylate (I) (3.0 mol) was added, followed by tris(dibenzylideneacetone)-dipalladium (O) (7.5 mol%) and tris-(2-furanyl)-phosphine (22.5 mol%). The mixture was placed in a preheated oil bath at 55 °C for 20 to 120 min until LC-MS indicated the consumption of boric acid. The mixture was cooled and then concentrated under reduced pressure. The residue was dissolved in ACN:H2O 1:1 (or 4:1 for less polar analogues), filtered through a PTFE filter (45 µm), and purified by RP-HPLC with an elution gradient of 15% to 85% ACN in H2O (+0.1% TFA).

[0086] 1 H NMR (300 MHz, CD3CN) δ 7.82 (s, 2H), 7.36 (d, J = 4.3 Hz, 2H), 6.71 (p, J = 2.2 Hz, 1H), 6.60 – 6.54 (m, 2H), 5.90 (hept, J = 6.3 Hz, 1H), 4.09 (dt, J = 23.1, 6.8 Hz, 4H), 3.19 (t, J = 2.1 Hz, 2H), 3.12 (t, J = 2.4 Hz, 2H).

[0087] Step 5) 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester The alkenyl-BODIPY compound 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester) was dissolved in anhydrous methanol, and Pd / C (20 mol%) was added. Triethylsilane (20 equivalents) was added dropwise at 0 °C. The reaction was heated to room temperature, and after completion (controlled by LC-MS, 60 min), a few drops of water were added, and Pd / C was removed by filtration through a PTFE filter (22 µm). The solution was concentrated under reduced pressure and purified by RP-HPLC to give the title compound (23 mg, 88%).

[0088] HRMS (ESI) m / z [M+H] + :C 20 H 18 The calculated value of BF8N3O2 is 494.1401, and the measured value is 494.1412.

[0089] 1 H NMR (600 MHz, CD3CN) δ 7.85 (s, 2H), 7.48 (d, J = 4.3 Hz, 2H), 6.63– 6.59 (m, 2H), 5.88 (hept, J = 6.4 Hz, 1H), 4.09 – 3.92 (m, 4H), 3.06 (d, J = 7.5 Hz, 2H), 2.50 (hept, J = 8.0 Hz, 1H), 2.28 – 2.23 (m, 2H), 2.05 – 2.00 (m, 2H).

[0090] Example 2 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid[ 18 F]-1,1,1,3,3,3-hexafluoropropane-2-ester 1,1,1,3,3,3-hexafluoropropane-2-ester (Example 1) of 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid was dissolved in anhydrous MeCN to prepare a stock solution with a concentration of 3 mg / mL. On the day of the experiment, 6 μL of the stock solution was mixed with 7.1 μmol SnCl4 in a reaction vial. 18 O(p,n) 18 The F reaction, using a Cyclone 18 / 9 cyclotron (18 MeV; IBA, Belgium), produces fluorine-18, containing [ 18 [F]fluoride ions were collected in aqueous solution on a QMA column (Waters SepPak Accell QMA carbonate column). The column was first washed with approximately 5 mL of Milli-Q water, then eluted with a solution of 75 mM tetraethylammonium bicarbonate in MeCN / H₂O (v / v = 4 / 5). After azeotropic drying with 1.0 mL × 2 MeCN, the residue was dissolved in 1 mL of anhydrous MeCN. Equal parts of [ 18 [F] Fluoride (approximately 100 μL) was partitioned into a reaction vial for radiolabeling. After shaking for 10 min at room temperature, the reaction was quenched with 10 mL of 0.1% TFA in Milli-Q water. The resulting mixture was passed through a pretreated MCX column and washed with 3 mL of Milli-Q water. Subsequently, the product was eluted with 1 mL of EtOH. The purity of the product was analyzed using an Agilent 1100 Series HPLC system equipped with a UV detector and a GabiStar radiodetector (Raytest) using an ACE XDB-C18 Zobrax column (75 mm × 4.6 mm, 3.5 μm). Mobile phases A and B were Milli-Q water (v / v) containing 0.1% H3PO4 and acetonitrile, respectively. A gradient method was used: 0.0–6.0 min, 5–60% B; 6.0–9.0 min, 60–95% B; 9.0–10.0 min, 95% B; 10.0–11.0 min, 95–5% B; wavelength 489 nm; flow rate 1 mL / min. The radioactive material was identified by co-injection with a reference compound (see Figure 1).

[0091] Example 3: 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid.

[0092] 5,5-Difluoro-10-(methylthio)-3-(1H-pyrrolo-2-yl)-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextriene Thiomethyl BODIPY (71.4 mg, 300 µmol) was dissolved in 2.5 mL of freshly distilled pyrrole and placed in a 10 mL sealed microwave-safe vial. The solution was heated to 150 °C under MW radiation in air for 60 minutes. The vial was then opened, and the reaction status was checked by LC-MS. This process was repeated until LC-MS indicated the formation of 3,8-bis(pyrrole-2-yl)BODIPY (m / z = 323 [M+H]) after 2 to 3 hours. + The mixture was concentrated under reduced pressure, and the resulting residue was purified by RP-HPLC from 50 to 95% ACN:H2O (+0.1% TFA). The title compound (8 mg, 9%) was isolated as a deep purple amorphous solid.

[0093] HRMS (ESI) m / z [M+H] + :C 14 H 12 The calculated value for BF2N3S is 302.0849, and the measured value is 302.0843.

[0094] 1 H NMR (300 MHz, CDCl3) δ 10.51 (s, 1H), 7.62 – 7.54 (m, 2H), 7.23 –7.19 (m, 2H), 7.18 (d, J = 4.1 Hz, 1H), 7.10 – 7.02 (m, 1H), 6.93 (d, J = 4.8Hz, 1H), 6.47 (t, J = 3.2 Hz, 1H), 6.43 – 6.39 (m, 1H), 2.70 (s, 3H).

[0095] Steps one through three are the same as those described in Example 1.

[0096] Step 4) 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester According to step 4 of Example 1, the title compound was synthesized from ((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid (7.9 mg, 19.8 µmol, 1.1 equivalents) and 5,5-difluoro-10-(methylthio)-3-(1H-pyrrolo-2-yl)-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohexanetriene (5.5 mg, 18 µmol, 1.0 equivalents). The product (5.9 mg, 59%) was obtained as a deep purple amorphous solid.

[0097] 1 H NMR (300 MHz, CDCl3) δ 10.50 (s, 1H), 7.59 (s, 1H), 7.21 – 7.09 (m,2H), 7.02 (ddd, J = 3.9, 2.5, 1.4 Hz, 1H), 6.90 (d, J = 4.7 Hz, 1H), 6.87 (d,J = 4.0 Hz, 1H), 6.48 (t, J = 2.3 Hz, 1H), 6.44 (dd, J = 3.9, 2.2 Hz, 1H), 6.40 (dq, J = 4.2, 2.2 Hz, 1H), 5.63 (hept, J = 6.2 Hz, 1H), 4.26 – 4.06 (m,4H), 3.17 (d, J = 2.3 Hz, 2H), 3.03 (d, J = 1.9 Hz, 2H).

[0098] Step 5) 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester According to step 5 of Example 1, the title compound was synthesized from 1,1,1,3,3,3-hexafluoropropane-2-ester (5.6 mg, 10 µmol) of 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid. It was obtained by RP-HPLC purification as a purple-red amorphous solid, exhibiting red fluorescence in solution (3 mg, 54%).

[0099] HRMS (ESI) m / z [M+H] + :C 24 H 21 The calculated value of BF8N4O2 is 559.1666, and the measured value is 559.1651.

[0100] 1 H NMR (300 MHz, CD3CN) δ 10.42 (s, 1H), 7.58 (s, 1H), 7.55 (d, J =4.8 Hz, 1H), 7.27 (q, J = 2.3 Hz, 1H), 7.18 (dt, J = 3.8, 1.2 Hz, 1H), 7.14(d, J = 3.6 Hz, 1H), 7.08 (d, J = 4.8 Hz, 1H), 6.50 (dd, J = 3.9, 2.2 Hz,1H), 6.39 (dt, J = 4.2, 2.3 Hz, 1H), 5.89 (hept, J = 6.2 Hz, 1H), 4.00 (t, J= 20.1 Hz, 4H), 2.97 (d, J = 7.4 Hz, 2H), 2.50 (hept, J = 7.8 Hz, 1H), 2.32 –2.19 (m, 2H), 2.08 – 1.95 (m, 2H).

[0101] Example 4: 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((5,5-difluoro-3-methyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid

[0102] Steps one through three are the same as those described in Example 1.

[0103] Step 4) 6-((5,5-difluoro-3-methyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester According to Step 4 of Example 1, the title compound was synthesized from ((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid (25.3 mg, 72.8 µmol, 1.0 equivalent) and 5,5-difluoro-3-methyl-10-(methylthio)-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohexanetriene (18.4 mg, 72.8 µmol, 1.0 equivalent), which was previously synthesized according to the literature (de J. Gómez-Infante, A. et al. Synthesis, Properties, and Functionalization of Nonsymmetric 8-MethylthioBODIPYs. European J. Org. Chem. 2016, Synthesized (5009–5023). The product (21.3 mg, 58%) was obtained as a deep red amorphous solid.

[0104] 1 H NMR (300 MHz, CD3CN) δ 7.64 (s, 1H), 7.32 (d, J = 4.4 Hz, 1H), 7.16 (d, J = 4.0 Hz, 1H), 6.60 (p, J = 2.2 Hz, 1H), 6.48 (dd, J = 4.4, 2.0 Hz,1H), 6.43 (d, J = 4.3 Hz, 1H), 5.90 (hept, J = 6.4 Hz, 1H), 4.20 – 3.99 (m,4H), 3.16 (q, J = 2.6 Hz, 2H), 3.04 (q, J = 2.6 Hz, 2H), 2.56 (s, 3H).

[0105] Step 5) 6-((5,5-difluoro-3-methyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester According to step 5 of Example 1, the title compound was synthesized from 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((5,5-difluoro-3-methyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid (21.3 mg, 42 µmol). It was obtained by RP-HPLC purification as an orange-red amorphous solid exhibiting green fluorescence in solution (16 mg, 75%).

[0106] 1 H NMR (300 MHz, CD3CN) δ 7.66 (s, 1H), 7.45 (d, J = 4.4 Hz, 1H), 7.28 (d, J = 4.1 Hz, 1H), 6.51 (dd, J = 4.4, 2.0 Hz, 1H), 6.47 (d, J = 4.4 Hz,1H), 5.89 (hept, J = 6.4 Hz, 1H), 3.99 (t, J = 20.2 Hz, 4H), 2.97 (d, J = 7.5Hz, 2H), 2.56 (s, 3H), 2.47 (p, J = 8.4 Hz, 1H), 2.23 (ddt, J = 9.5, 7.6, 2.3Hz, 2H), 2.07 – 1.94 (m, 2H).

[0107] Example 5: 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((5,5-difluoro-3,7-dimethyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid.

[0108] Steps one through three are the same as those described in Example 1.

[0109] Step 4) 6-((5,5-difluoro-3,7-dimethyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester According to Step 4 of Example 1, the title compound was synthesized from ((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid (52.1 mg, 150 µmol, 1.5 equivalents) and 5,5-difluoro-3,7-dimethyl-10-(methylthio)-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohexanetriene (26.6 mg, 100 µmol, 1.0 equivalents), which was previously synthesized according to the literature (de J. Gómez-Infante, A. et al. Synthesis, Properties, and Functionalization of Nonsymmetric 8-MethylthioBODIPYs. European J. Org. Chem. 2016, Synthesized (5009–5023). The product (12.4 mg, 24%) was obtained as a deep red amorphous solid.

[0110] 1 H NMR (300 MHz, CD3CN) δ 7.14 (d, J = 4.1 Hz, 2H), 6.52 (p, J = 2.2Hz, 1H), 6.33 (d, J = 4.1 Hz, 2H), 6.00 – 5.81 (m, 1H), 4.07 (dt, J = 21.0,7.4 Hz, 4H), 3.13 (d, J = 2.1 Hz, 2H), 3.00 (d, J = 2.6 Hz, 2H), 2.53 (s,6H).

[0111] Step 5: 6-((5,5-difluoro-3,7-dimethyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester According to step 5 of Example 1, the title compound was synthesized from 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-((5,5-difluoro-3,7-dimethyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid (12.5 mg, 24 µmol). It was obtained by RP-HPLC purification as an orange-red amorphous solid, exhibiting green fluorescence in solution (9.1 mg, 72%).

[0112] 1 H NMR (300 MHz, CD3CN) δ 7.27 (d, J = 4.2 Hz, 2H), 6.36 (d, J = 4.2Hz, 2H), 5.89 (hept, J = 6.5 Hz, 1H), 3.99 (t, 4H), 2.91 (d, J = 7.4 Hz, 2H), 2.52 (s, 6H), 2.42 (dq, J = 15.9, 8.0 Hz, 1H), 2.22 (ddt, J = 9.6, 7.6, 2.3Hz, 2H), 2.00 (d, J = 10.0 Hz, 2H).

[0113] Example 6 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester

[0114] (R)-1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-ol According to the literature (McAllister, LA et al. Discovery of Trifluoromethyl Glycol Carbamates as Potent and Selective Covalent Monoacylglycerol Lipase (MAGL) Inhibitors for Treatment of Neuroinflammation. J. Med. Chem. 61, 3008–3026), (4-methoxyphenyl)methanol (CAS 105-13-5, 777 mg, 5.625 mmol, 1.5 equivalents) was slowly added to a 0°C solution of NaH (60 wt% in mineral oil, 450 mg, 11.25 mmol, 3 equivalents) in tetrahydrofuran (10 mL). After stirring the reaction mixture at 0 °C for 60 min, (2R)-2-(trifluoromethyl)ethylene oxide (prepared by hydrolytic kinetic resolution from a racemic mixture (Schaus et al. 2002), 420 mg, 3.75 mmol) was added to 2 mL of THF, and the mixture was heated to ambient temperature and stirred for 40 h. The reaction mixture was then cooled to 0 °C and quenched by adding water. The solution was separated by water and EtOAc. The aqueous phase was extracted two or more times with EtOAc, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The title compound was purified by rapid chromatography with an elution gradient of 0–20% EtOAc in cyclohexane. The product (635 mg, 68%) was given as a pale yellow oil.

[0115] 1 H NMR (300 MHz, CDCl3) δ 7.30 – 7.21 (m, 2H), 6.95 – 6.85 (m, 2H), 4.53 (d, J = 1.2 Hz, 2H), 4.12 (d, J = 5.7 Hz, 1H), 3.82 (s, 3H), 3.76 – 3.57(m, 2H), 2.88(s, 1H).

[0116] Step 2) 6-((4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-ester 6-((4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (see Example 1, Step 1) (80.4 mg, 240 µmol) was deprotected by BOC using a TFA / DCM 1:4 mixture. Triethylamine (200 µl, 1.44 mmol) and bis(pentafluorophenyl)carbonate (CAS 59483-84-0, 136 mg, 346 µmol) were added to a solution of (R)-1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-ol (72 mg, 288 µmol) in anhydrous acetonitrile (2 mL). The reaction mixture was stirred at 0 °C for 1 hour and then heated to room temperature. The mixture was then treated with a solution of the previously deprotected boc intermediate and triethylamine (167 µL, 1.2 mmol) in anhydrous acetonitrile (2 mL). The reaction mixture was stirred at room temperature for 16 hours, followed by concentration under reduced pressure. The crude product was purified by silica gel chromatography with an elution gradient of 0–20% EtOAc in cyclohexane. The title compound (102 mg, 82%) was given as a colorless amorphous solid.

[0117] HRMS (ESI) m / z [M+H] + :C 25 H 33 The calculated value of BF3NO6 is 510.2389, and the measured value is 510.2405.

[0118] 1 H NMR (300 MHz, CDCl3) δ 7.25 (d, J = 8.9 Hz, 2H), 6.93 – 6.84 (m,2H), 5.38 (qd, J = 7.1, 4.0 Hz, 1H), 5.21 (t, J = 2.3 Hz, 1H), 4.50 (q, J =11.7 Hz, 2H), 4.19 – 3.99 (m, 4H), 3.81 (s, 3H), 3.69 (qd, J = 11.2, 5.5 Hz, 2H), 3.11 (d, J = 2.5 Hz, 2H), 2.96 (s, 2H), 1.24 (s, 12H).

[0119] Step 3) (R)-((2-(((1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid The title compound was obtained from the intermediate (102 mg, 200 µmol) of step 2, as in step 3 of Example 1. The crude product (H2O / EtOAc) obtained after liquid extraction was used without further purification.

[0120] Step 4) 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-ester According to GP6, the title compound was synthesized from crude (R)-((2-(((1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid (32 mg, approx. 80% purity, approx. 67.5 µmol, 1.5 equivalents) and 8-thiomethyl-bodipy (10.7 mg, 45 µmol). The title compound (23 mg, 89%) was obtained as a deep red amorphous solid after RP-HPLC.

[0121] HRMS (ESI) m / z [M+H] + :C 28 H 27 The calculated value of BF5N3O4 is 574.2051, and the measured value is 574.2038.

[0122] 1 H NMR (300 MHz, CDCl3) δ 7.84 (s, 2H), 7.25 – 7.19 (m, 2H), 7.16 (d,J = 4.2 Hz, 2H), 6.88 (s, 2H), 6.56 (tt, J = 2.2, 1.3 Hz, 1H), 6.51 (dd, J =4.2, 1.7 Hz, 2H), 5.39 (pd, J = 7.0, 3.8 Hz, 1H), 4.49 (q, J = 11.7 Hz, 2H), 4.16 – 4.01 (m, 4H), 3.79 (s, 3H), 3.70 (td, J = 12.8, 5.8 Hz, 2H), 3.16 (s,2H), 3.07 (s, 2H).

[0123] Step 5) 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-ester (11.5 mg, 20 µmol) was dissolved in DCM (2 mL), and H2O (0.1 mL) and DDQ (CAS 84-58-2, 6.8 mg, 30 µmol) were added. The mixture was stirred vigorously at ambient temperature for 18 hours. The mixture was separated by distillation with saturated sodium bicarbonate solution and DCM. The aqueous layer was extracted twice with DCM, and the combined organic phases were dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by RP-HPLC to obtain the title compound (5.42 mg, 60%) as a deep red amorphous solid.

[0124] 1 H NMR (300 MHz, CD3CN) δ 7.82 (s, 2H), 7.36 (d, J = 4.2 Hz, 2H), 6.71 (p, J = 2.2 Hz, 1H), 6.57 (dd, J = 4.3, 1.7 Hz, 2H), 5.17 (pd, J = 7.2, 3.9Hz, 1H), 4.15 – 3.94 (m, 4H), 3.80 (dd, J = 12.4, 4.0 Hz, 1H), 3.69 (dd, J =12.4, 6.8 Hz, 1H), 3.18 (q, J = 2.7 Hz, 2H), 3.10 (q, J = 2.7 Hz, 2H).

[0125] Step 6) AH222 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester The title compound was synthesized from the intermediate (5.4 mg, 12 µmol) in step 5 of Example 1. The product was obtained as an orange solid by RP-HPLC, exhibiting green fluorescence in solution (5.4 mg, quantitative yield).

[0126] HRMS (ESI) m / z [M+H] + :C 20 H 21 The calculated value of BF5N3O3 is 456.1632, and the measured value is 456.1637.

[0127] 1 H NMR (300 MHz, CD3CN) δ 7.85 (s, 2H), 7.49 (d, J = 4.3 Hz, 2H), 6.65– 6.57 (m, 3H), 5.16 (pd, J = 7.2, 3.9 Hz, 1H), 4.07 – 3.85 (m, 4H), 3.85 –3.62 (m, 2H), 3.05 (d, J = 7.5 Hz, 2H), 2.49 (tt, J = 8.9, 7.3 Hz, 1H), 2.30– 2.16 (m, 1H), 2.12 – 1.95 (m, 2H).

[0128] Example 7 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester

[0129] Steps one and two are the same as in Example 6.

[0130] Step 3) (R)-((2-(((1,1,1-trifluoro-3-hydroxypropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid As described in Step 4 of Example 6, the title compound was synthesized by PMB deprotection of 6-((4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-((4-methoxybenzyl)oxy)propane-2-ester (46 mg, 90 µmol) with DDQ. The crude residue after liquid-liquid extraction was hydrolyzed with pinacol ester (Bpin) under the conditions of Step 3 of Example 1, without prior purification of the crude product. The title compound (16.9 mg, 61%, in 2 steps) was then purified and separated as a colorless solid by RP-HPLC.

[0131] 1 H NMR (300 MHz, MeOD) δ 5.34 (p, J = 2.3 Hz, 1H), 5.21 (pd, J = 7.2,3.8 Hz, 1H), 4.15 (s, 1H), 4.10 – 4.01 (m, 3H), 3.85 (dd, J = 12.4, 3.8 Hz,1H), 3.73 (dd, J = 12.4, 7.2 Hz, 1H), 3.16 – 3.04 (m, 2H), 3.02 – 2.94 (m,2H).

[0132] Step 4) 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester According to step 4 of Example 1, the title compound was synthesized from (R)-((2-(((1,1,1-trifluoro-3-hydroxypropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-ylidene)methyl)boronic acid (10.7 mg, 34.5 µmol, 1.5 equivalents) and 5,5-difluoro-10-(methylthio)-3-(1H-pyrrolo-2-yl)-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohexanetriene (7.0 mg, 23 µmol, 1.0 equivalents). The product (7.26 mg, 61%) was obtained as a deep purple amorphous solid.

[0133] 1H NMR (300 MHz, CD3CN) δ 10.44 (s, 1H), 7.56 (d, J = 2.1 Hz, 1H), 7.41 (d, J = 4.8 Hz, 1H), 7.28 (td, J = 2.8, 1.4 Hz, 1H), 7.18 (ddd, J = 4.0,2.6, 1.4 Hz, 1H), 7.05 (d, J = 4.8 Hz, 1H), 7.02 (d, J = 4.1 Hz, 1H), 6.57(p, J = 2.1 Hz, 1H), 6.47 (dd, J = 4.0, 2.3 Hz, 1H), 6.40 (dt, J = 4.2, 2.4Hz, 1H), 5.17 (pd, J = 7.2, 4.0 Hz, 1H), 4.10 – 3.99 (m, 4H), 3.80 (dd, J =12.3, 3.9 Hz, 1H), 3.69 (dd, J = 12.4, 6.7 Hz, 1H), 3.15 (d, J = 2.2 Hz, 2H),3.02 (d, J = 2.6 Hz, 2H).

[0134] Step 5) 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester According to Example 1, the title compound was synthesized from 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester (7.28 mg, 14 µmol). It was obtained by RP-HPLC as a deep purple-red solid with red fluorescence in solution (3.64 mg, 50%).

[0135] HRMS (ESI) m / z [M+H] + :C 24 H 24 The calculated value of BF5N4O3 is 521.1898, and the measured value is 521.1951.

[0136] 1 H NMR (600 MHz, CD3CN) δ 10.43 (s, 1H), 7.58 (s, 1H), 7.55 (d, J =4.8 Hz, 1H), 7.27 (td, J = 2.8, 1.4 Hz, 1H), 7.19 – 7.17 (m, 1H), 7.15 (d, J = 3.9 Hz, 1H), 7.07 (d, J = 4.8 Hz, 1H), 6.50 (dd, J = 4.0, 2.2 Hz, 1H), 6.40 (dt, J = 4.4, 2.4 Hz, 1H), 5.16 (pd, J = 7.1, 3.9 Hz, 1H), 4.09 – 3.84 (m,4H), 3.79 (dd, J = 12.3, 3.9 Hz, 1H), 3.69 (dd, J = 12.6, 6.8 Hz, 1H), 2.98 (d, J = 7.5 Hz, 2H), 2.50 (hept, J = 8.0 Hz, 1H), 2.28 – 2.21 (m, 2H), 2.02(dd, J = 12.2, 8.8 Hz, 2H).

[0137] Example 8 6-(2-(2-(3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-3-yl)propamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester

[0138] Step 1 is the same as Step 1 in Example 1.

[0139] Step 2) (2-(2-bromophenoxy)ethyl)carbamate benzyl ester 2-Bromophenol (CAS 95-56-7, 882 mg, 5.10 mmol) was dissolved in DMF (4 mL), Cs₂CO₃ (1 equivalent) was added, and the mixture was stirred at room temperature for five minutes. Next, 2-(((benzyloxy)carbonyl)amino)ethyl 4-methylbenzenesulfonic acid (CAS 93407-96-6, 1.6 equivalents) was added, and the mixture was stirred at 55 °C for 18 hours. The reaction was quenched by dilution with DCM (60 mL), extracted with 10% NaOH solution (2 x 40 mL), and washed with brine (50 mL). The organic layer was dried over MgSO₄, filtered, and concentrated. The crude product was purified by silica gel rapid chromatography (0 to 50% EtOAc in cyclohexane) to give the title compound.

[0140] 1 H NMR (300 MHz, CDCl3) δ 7.47 (dd, J = 7.9, 1.6 Hz, 1H), 7.33 – 7.13(m, 6H), 6.87 – 6.73 (m, 2H), 5.31 (s, 1H), 5.06 (s, 2H), 4.03 (t, J = 5.0Hz, 2H), 3.59 (q, J = 5.4Hz, 2H).

[0141] Step 3) 6-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester In a sealed vial under a nitrogen atmosphere, tert-butyl 6-((4,4,5,5-tetramethyl-1,3,2-dioxaborphane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylate (117 mg, 0.35 mmol), benzyl (2-(2-bromophenoxy)ethyl)carbamate (123 mg, 0.35 mmol), and Na₂CO₃ (74.2 mg, 0.70 mmol) were suspended in a 3:1 dioxane / water mixture (2.30 mL) previously purged with sonicated bubbling N₂. The mixture was purged with N₂ for 10 min. Then, Pd(dppf)Cl₂ (CAS72287-26-4) was added, and the mixture was purged for another 15 min. The reaction was heated to 55 °C and stirred at this temperature overnight. The reactants were then diluted with DCM (15 mL), extracted with saturated NaHCO3 aqueous solution (20 mL), and washed with brine (20 mL). The organic layer was dried over MgSO4, concentrated under reduced pressure, and purified by silica gel chromatography (0 to 60% EtOAc in cyclohexane) to give the title compound (97.0 mg, 0.20 mmol, 58%).

[0142] HRMS (ESI): C 28 H 34 N₂O₅Na [M+Na]⁺ Calculated value: 501.2360; Measured value: 501.2388.

[0143] Step 4) 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl-2-azaspiro[3.3]heptane-2-carboxylic acid The intermediate was synthesized by first deprotecting 6-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (67 mg, 200 µmol) by stirring in 20% TFA in DCM for 3 hours, followed by removal of any volatiles under reduced pressure. The deprotected amine was dissolved in 10 mL of anhydrous DCM and DIPEA (45 µl, 260 µmol) was added. The mixture was cooled to 0 °C and then a solution of ~50 wt% bis(1,1,1,3,3,3-hexafluoropropane-2-yl) carbonate in Et2O (1 equivalent, freshly prepared according to the literature) was added. The mixture was heated to room temperature and stirred for 3 h. The mixture was diluted with 10 mL of DCM and washed with saturated NaCl solution (5 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The title compound (60 mg, 35%) was purified by RP-HPLC to obtain a colorless solid.

[0144] 1 H NMR (600 MHz, CDCl3) δ 7.40 – 7.30 (m, 5H), 7.16 (t, J = 7.5 Hz, 2H), 6.93 (t, J = 7.5 Hz, 1H), 6.84 (d, J = 8.4 Hz, 1H), 6.49 (t, J = 2.6 Hz,1H), 5.65 (hept, J = 6.2 Hz, 1H), 5.18 (s, 1H), 5.12 (s, 2H), 4.15 (m, 4H),4.06 (t, J = 5.2 Hz, 2H), 3.64 (m, 2H), 3.21 (s, 2H), 3.10 (s, 2H).

[0145] Step 5) 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-(2-aminoethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylene)-2-azaspiro[3.3]heptane-2-carboxylic acid (68.7 mg, 120 mmol) was dissolved in EtOAc: tIn BuOH (4:1), Pd / C (20 mol%) was added, and the suspension was first degassed by ultrasound. Then, hydrogen gas was continuously bubbled into the suspension for 150 minutes at room temperature. The suspension was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound as a colorless, viscous solid in quantitative yield.

[0146] Step 6) 6-(2-(2-(3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-3-yl)propamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-3-yl)propionic acid (BODIPY-Fl, CAS 165599-63-3, 4.4 mg, 15 µmol) and HATU (6.8 mg, 18 µmol, 1.2 equivalents) were dissolved in anhydrous DMF, and 2,6-dimethylpyridine (3.2 mg, 2 equivalents) was added. The mixture was stirred at ambient temperature for 10 minutes. 6-(2-(2-aminoethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid (1,1,1,3,3,3-hexafluoropropane-2-ester (7.9 mg, 15 µmol)) was added, and the mixture was stirred for 18 hours. The mixture was concentrated under reduced pressure, and the residue was purified by RP-HPLC to give the title compound (6 mg, 56%) as an orange amorphous solid that fluoresces green in solution.

[0147] 1H NMR (600 MHz, CD3CN) δ 7.29 (s, 1H), 7.15 (td, J = 7.8, 1.8 Hz,1H), 7.07 (dd, J = 7.5, 1.7 Hz, 1H), 6.92 – 6.82 (m, 3H), 6.63 (s, 1H), 6.29(d, J = 4.0 Hz, 1H), 6.21 (s, 1H), 5.88 (hept, J = 6.4 Hz), 4.05 – 3.89 (m,6H), 3.54 (m, 2H), 3.17 (m, 2H), 2.63 (m, 2H), 2.57 (t, J = 7.5 Hz, 2H), 2.49(s, 3H), 2.41 – 2.38 (m, 1H), 2.25 (s, 3H), 2.23 – 2.18 (m, 2H), 1.92 – 1.87(m, 2H).

[0148] Example 9 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-(2-(4-((4-(dimethylamino)phenyl)but-1,3-diyne-1-yl)benzamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid Steps 1 through 5 are the same as in Example 8.

[0149] Step 6) According to Example 8, Step 6, the title compound (9 mg, 72%) was synthesized from 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-(2-aminoethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid (7.7 mg, 18 µmol, 1.0 equivalent) and 4-((4-(dimethylamino)phenyl)but-1,3-diyn-1-yl)benzoic acid (5.1 mg, 18 µmol) as a yellow solid.

[0150] 1H NMR (300 MHz, CDCl3) δ 7.72 (d, J = 8.3 Hz, 2H), 7.57 (d, J = 8.3Hz, 2H), 7.46 (d, J = 8.7 Hz, 2H), 7.37 (d, J = 4.4 Hz, 1H), 7.23 – 7.14 (m,1H), 7.06 (d, J = 7.3 Hz, 1H), 6.97 – 6.82 (m, 3H), 6.46 (s, 1H), 5.63 (p, J= 6.2 Hz, 1H), 4.17 (t, J = 5.2 Hz, 2H), 4.04 (d, J = 11.3 Hz, 2H), 3.96 (d,J = 13.1 Hz, 2H), 3.90 (q, J = 5.3 Hz, 2H), 3.04 (s, 6H), 2.68 (d, J = 7.2Hz, 2H), 2.50 – 2.39 (m, 1H), 2.26 (t, J = 9.9 Hz, 2H), 1.91 (t, J = 10.0 Hz, 2H).

[0151] Example 10 N-(10-(2-carboxy-5-((5-(2-((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-yl)methyl)phenoxy)pentyl)carbamoyl)phenyl)-7-(dimethylamino)-5,5-dimethyldibenzo[b,e]silicyclohexanetriene-3(5H)-ylidene)-N-methylmethylammonium

[0152] Step 1 is the same as Step 1 in Example 1.

[0153] Step 2) (5-(2-bromophenoxy)pentyl)carbamate benzyl ester According to Step 2 of Example 8, the title compound was synthesized from 2-bromophenol (2.4 mmol) and 5-(((benzyloxy)carbonyl)amino)pentyl 4-methylbenzenesulfonic acid (CAS 93066-51-4, 2 mmol). The title compound (51%) was isolated as a colorless oil.

[0154] 1¹H NMR (300 MHz, CDCl₃) δ 7.52 (dd, J = 7.9, 1.6 Hz, 1H), 7.39 – 7.20 (m, 6H), 6.94 – 6.71 (m, 2H), 5.10 (s, 2H, OCH₂), 4.78 (s, 1H, NHCbz), 4.01 (t, J = 6.2 Hz, 2H, OCH₂), 3.24 (q, J = 6.3 Hz, 2H, CH₂NHCbz), 1.86 (t, J = 6.8 Hz, 2H, CH₂ linker), 1.76 – 1.48 (m, 4H, 2 x CH₂ linker). 13 C NMR (75 MHz, CDCl3) δ 156.4, 155.3 (C=O Cbz, C Ar -O)

[0155] Step 3) 6-(2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester According to step 3 of Example 8, the title compound was synthesized from (5-(2-bromophenoxy)pentyl)carbamate (640 mg, 1.63 mmol) and 6-((4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylate tert-butyl ester (601 mg, 1.8 mmol). A pale yellow oil (571 mg, 67%) was obtained.

[0156] HRMS (ESI): C 31 H 41 N₂O₅ [M+H] + Calculated value: 521.3010; Measured value: 521.3000

[0157] Step 4) 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzylene)-2-azaspiro[3.3]heptane-2-carboxylic acid The title compound was synthesized from tert-butyl 6-(2-((5-((((benzyloxy)carbonyl)amino)pentyl)oxy)benzylene)-2-azaspiro[3.3]heptane-2-carboxylate according to step 4 of Example 8. A colorless amorphous solid (81 mg, 0.14 mmol, 47%) was obtained.

[0158] 1 H NMR (300 MHz, CDCl3) δ 7.36 – 7.19 (m, 5H), 7.16 – 6.99 (m, 2H), 6.85 – 6.66 (m, 2H), 6.44 (t, J = 2.4 Hz, 1H), 5.57 (p, J = 6.2 Hz, 1H), 5.01(s, 2H), 4.10 – 3.97 (m, 4H), 3.87 (t, J = 6.4 Hz, 2H), 3.21 – 3.08 (m, 4H), 3.00 (d, J = 3.7 Hz, 2H), 1.74 (p, J = 6.6 Hz, 2H), 1.56 – 1.36 (m, 4H).

[0159] Step 5) 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-((5-aminopentyl)oxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid According to step 5 of Example 8, the title compound was synthesized from 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-((5-((((benzyloxy)carbonyl)amino)pentyl)oxy)benzylene)-2-azaspiro[3.3]heptane-2-carboxylic acid. The crude product, a colorless oil, can be used in subsequent steps without further purification.

[0160] Step 6) N-(10-(2-carboxy-5-((5-(2-((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-yl)methyl)phenoxy)pentyl)carbamoyl)phenyl)-7-(dimethylamino)-5,5-dimethyldibenzo[b,e]silicyclohexanetriene-3(5H)-ylidene)-N-methylmethylammonium The title compound was synthesized from 1,1,1,3,3,3-hexafluoropropane-2-ester of 6-(2-((5-aminopentyl)oxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid and silirodamine-COOH (Spirochrome AG, product number SC004) as a blue amorphous solid (5.7 mg, 63%).

[0161] HRMS (ESI): C 49 H 55 F6N4O6Si [M+H] +Calculated value: 937.3790; Measured value: 937.3787.

Claims

1. A compound of formula (I) (I) Or its pharmaceutically acceptable salt, wherein: X is CH and Y is CH2; or X and Y together form the group CH=C; R 1 Selected from: ； R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; and R 2 Selected from: , and ; R 3a and R 3b Each is independently selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; The reporting unit is selected from: ; ;and ; L is selected from –(CH2) p –, –(CH2)2-NHC(O)-CH2–, –CH2-NHC(O)-(CH2)2–, –((CH2)2O) q -(CH2)2– and –(CH2)2-(O(CH2)2) q –; n and m are each independently 1 or 2; p is selected from 1, 2, 3, 4, and 5; q is selected from 1, 2, and 3; and r is selected from 1, 2, 3, and 4.

2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein X is CH and Y is CH2.

3. The compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R 1 for , where R 1a As defined in claim 1.

4. The compound of formula (I) according to claim 3, or a pharmaceutically acceptable salt thereof, wherein R 1a It is a halogenated-C1-C6-alkyl group.

5. The compound of formula (I) according to claim 3, or a pharmaceutically acceptable salt thereof, wherein R 1a Selected from CF3 and hydroxymethyl.

6. The compound of formula (I) according to claim 3, or a pharmaceutically acceptable salt thereof, wherein R 1a It is CF3.

7. The compound of formula (I) according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , and ; Where R 3a R 3b R 3c m, n, and r are as defined in claim 1.

8. The compound of formula (I) according to claim 7, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , and ;in: R 3a Selected from hydrogen and C1-C6-alkyl; R 3b Selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; m is 1 or 2; and n is 1.

9. The compound of formula (I) according to claim 8, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , and ;in: R 3a Selected from hydrogen and methyl; R 3b Selected from hydrogen, methyl, and pyrrole groups; R 3c Selected from F and 18 F; m is 1 or 2; and n is 1.

10. The compound of formula (I) according to claim 9, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , , , and ;in: R 3c Selected from F and 18 F.

11. The compound of formula (I) according to claim 10, or a pharmaceutically acceptable salt thereof, wherein R 2 Selected from: , , , and ;in: R 3c Selected from F and 18 F.

12. The compound of formula (I) according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein R 3c It is F.

13. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: X is CH and Y is CH2; X and Y together form the group CH=C; R 1 for ; R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; R 2 Selected from: , and ;in: R 3a Selected from hydrogen and C1-C6-alkyl; R 3b Selected from hydrogen, C1-C6-alkyl, and 5- to 6-membered heteroaryl groups; R 3c Selected from F and 18 F; m is 1 or 2; and n is 1.

14. The compound of formula (I) according to claim 13, or a pharmaceutically acceptable salt thereof, wherein: X is CH and Y is CH2; R 1 for ; R 1a Selected from halogenated-C1-C6-alkyl and hydroxy-C1-C6-alkyl; R 2 Selected from: , , , , , and ;in: R 3c Selected from F and 18 F.

15. The compound of formula (I) according to claim 14, or a pharmaceutically acceptable salt thereof, wherein: X is CH and Y is CH2; R 1 for ; R 1a Selected from CF3 and hydroxymethyl; R 2 Selected from: , , , , , and ;in: R 3c It is F.

16. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) is selected from: 6-((5,5-difluoro-5H-4λ4,5λ4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid[ 18 F]-1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3-methyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-3,7-dimethyl-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-((5,5-difluoro-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester; 6-((5,5-difluoro-3-(1H-pyrrolo-2-yl)-5H-4l4,5l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-10-yl)methyl)-2-azaspiro[3.3]heptane-2-carboxylic acid(R)-1,1,1-trifluoro-3-hydroxypropane-2-ester; 6-(2-(2-(3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborhexacyclohextrien-3-yl)propamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; 6-(2-(2-(4-((4-(dimethylamino)phenyl)but-1,3-diyne-1-yl)benzamido)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carboxylic acid 1,1,1,3,3,3-hexafluoropropane-2-ester; and N-(10-(2-carboxy-5-((5-(2-((2-(((1,1,1,3,3,3-hexafluoropropane-2-yl)oxy)carbonyl)-2-azaspiro[3.3]heptane-6-yl)methyl)phenoxy)pentyl)carbamoyl)phenyl)-7-(dimethylamino)-5,5-dimethyldibenzo[b,e]silicyclohexanetriene-3(5H)-ylidene)-N-methylmethylammonium.

17. The compound of formula (I) according to any one of claims 1 to 16, used for a study of monoacylglycerol lipase (MAGL) occupancy.

18. The compound of formula (I) according to any one of claims 1 to 16, for diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals.

19. The compound of formula (I) according to any one of claims 1 to 16, used to generate equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

20. Use of the compound of formula (I) according to any one of claims 1 to 16 in a study of monoacylglycerol lipase (MAGL) occupancy.

21. Use of the compound of formula (I) according to any one of claims 1 to 16 in diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals.

22. Use of the compound of formula (I) according to any one of claims 1 to 16 for generating equilibrium and kinetic binding data of monoacylglycerol lipase (MAGL).

23. A method for studying the occupancy of monoacylglycerol lipase (MAGL), the method comprising contacting MAGL with a compound of formula (I) according to any one of claims 1 to 16.

24. A method for diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals, the method comprising contacting MAGL with a compound of formula (I) according to any one of claims 1 to 16.

25. A method for generating equilibrium and kinetic binding data of monoacylglycerol lipase (MAGL), the method comprising contacting MAGL with a compound of formula (I) according to any one of claims 1 to 16.

26. The present invention as described above.

Images