Fluorescent probe for MAGL

Abstract

The present invention relates to a compound represented by general formula (I) or (II): TIFF2025530870000170.tif101165 (Wherein A, L, n and R 1 ~R 3 is as defined herein) or a pharmaceutically acceptable salt thereof, compositions comprising said compounds, methods for making said compounds, and methods for using said compounds.

Description

[Technical Field]

[0001] The present invention relates to organic compounds useful as fluorescent probes for monoacylglycerol lipase (MAGL). [Background technology]

[0002] Fluorescent imaging probes have emerged as high-resolution tools for investigating protein localization in live cells, including expression levels and protein distribution, structure, dynamics, and function in health and disease (LAStoddart, LEKilpatrick, SJBriddon, SJHill, Neuropharmacology 2015, 98, 48-57). Such probes can be applied, for example, to cellular trafficking studies using flow cytometry fluorescence-activated cell sorting (FACS) experiments or confocal live-cell imaging. Furthermore, fluorescent imaging probes enable real-time monitoring of ligand-receptor interactions and visualization of proteins with high spatiotemporal precision (AJ Vernall, SJ Hill, B. Kellam, Br. J. Pharmacol. 2014, 171, 1073-1084; C. Iliopoulos-Tsoutsouvas, R.N. Kulkarni, A. Makriyannis, S.P. Nikas, Expert Opin. Drug Discov. 2018, 13, 933-947). Furthermore, such probes offer the possibility of generating equilibrium and kinetic binding data in a high-throughput manner without handling radioactive materials, for example, using time-resolved fluorescence resonance energy transfer (TR-FRET). Fluorescent imaging probes can also be useful in supporting the translation of preclinical pharmacological animal data to the clinic and can be applied to dose selection in humans. They can be used, for example, as markers of target engagement via the generation of ex vivo quantitative receptor binding data in whole blood. Depending on the respective application, fluorescent imaging probes must meet certain criteria, including affinity, selectivity and specificity for the respective target, favorable photophysical properties, and applicability across different techniques and cell types. Summary of the Invention

[0003] In a first aspect, the present invention provides a compound of formula (I) or (II): [ka] [ka] (Wherein A, L, n and R 1 ~R 3 is as defined herein) or a pharmaceutically acceptable salt thereof.

[0004] In a further aspect, the present invention provides a method for preparing the compounds of formula (I) or (II).

[0005] In a further aspect, the present invention provides a compound of formula (I) or (II) as described herein when prepared according to the methods described herein.

[0006] In a further aspect, the present invention provides the use of a compound of formula (II) as described herein for the preparation of a fluorescent probe of formula (I) as described herein.

[0007] In a further aspect, the present invention provides a method of testing monoacylglycerol lipase (MAGL) occupancy, comprising contacting MAGL with a compound of formula (I) described herein.

[0008] In a further aspect, the present invention provides a method for diagnostic imaging of monoacylglycerol lipase (MAGL) in a mammal, comprising contacting the MAGL with a compound of formula (I) described herein.

[0009] In a further aspect, the present invention provides a method for generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL), comprising contacting MAGL with a compound of formula (I) described herein. DETAILED DESCRIPTION OF THE INVENTION

[0010] definition It should be understood that any feature, integer, characteristic, compound, chemical moiety, or group described in connection with a particular aspect, embodiment, or example of the invention is applicable to any other aspect, embodiment, or example described herein, except where inconsistent therewith. All features disclosed herein (including any accompanying claims, abstract, and drawings), and / or all steps of any method or process so disclosed, may be combined in any combination, except combinations in which at least some of such features and / or steps are mutually exclusive. The invention is not limited to the details of any of the foregoing embodiments. The invention extends to any novel one or any novel combination of features disclosed herein (including any accompanying claims, abstract, and drawings), or any novel one or any novel combination of steps of any method or process so disclosed.

[0011] The term "heteroaryl" refers to a monovalent or polyvalent monocyclic, bicyclic, or tricyclic, preferably bicyclic, ring system having a total of 5 to 14 ring members, preferably 5 to 12 ring members, and more preferably 5 to 10 ring members, wherein at least one ring in the system is aromatic and at least one ring in the system contains one or more heteroatoms. Preferably, "heteroaryl" refers to a 5- to 10-membered heteroaryl containing 1, 2, 3, or 4 heteroatoms independently selected from O, S, and N. Most preferably, "heteroaryl" refers to a 5- to 10-membered heteroaryl containing 1 to 2 heteroatoms independently selected from O, S, and N.Some non-limiting examples of heteroaryl include spiro[cyclopropane-1,3′-indoline] (e.g., spiro[cyclopropane-1,3′-indoline]-1′-yl), 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazin-2-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrimidin-6-yl, indol-1-yl, 1H-indol-2-yl, 1H-indol-3-yl, 1H-indol-4-yl, 1H-indol-5-yl, 1H-indol-6-yl, 1H-indol-7-yl, 1,2-benzoxazol-3-yl, 1,2-benzoxazol-4-yl, 1,2-benzoxazol-5-yl, 1,2-benzoxazol-6-yl, 1,2-benzoxazol-7-yl, 1H-indazol-3-yl, 1H-indazol-4-yl, 1H-indazol-5-yl, 1H-indazol-6-yl, 1H-indazol-7-yl, pyrazol-1-yl, 1H-pyrazol-3-yl, 1H- pyrazol-4-yl, 1H-pyrazol-5-yl, pyrazolo[1,5-a]pyridine, 2H-pyrazolo[4,3-b]pyridine, [1,2,4]triazolo[1,5-a]pyridine, 1H-pyrrolo[2,3-b]pyridine, imidazol-1-yl, 1H-imidazol-2-yl, 1H-imidazol-4-yl, 1H-imidazol-5-yl, oxazol-2-yl, oxazol-4-yl, oxazol-5-yl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, Examples include thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, pyridazin-3-yl, pyridazin-4-yl, 1,2,4-triazol-4-yl, 1,2,4-triazol-1-yl, 4H-1,2,4-triazol-3-yl, 4,5,6,7-tetrahydroindazol-2-yl, 6,7-dihydro-4H-pyrano[4,3-c]pyrazol-2-yl, thiazolyl, benzofurazan-4-yl, tetrazolyl, isoxazolyl, pyrrolyl, and morpholinyl.

[0012] The term "aryl" refers to a group of rings containing a total of 6 to 10 ring members ("C6-C 10"aryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic ring system having at least one ring in the system that is aromatic. Some non-limiting examples of aryl include phenyl and 9H-fluorenyl (e.g., 9H-fluoren-9-yl). A particularly preferred, but non-limiting example of aryl is phenyl.

[0013] The term "alkyl" refers to a monovalent or polyvalent, e.g., monovalent or divalent, straight-chain or branched saturated hydrocarbon group having 1 to 12 carbon atoms. In some preferred embodiments, the alkyl group contains 1 to 6 carbon atoms, e.g., 1, 2, 3, 4, 5, or 6 carbon atoms ("C1-C6-alkyl"). In other embodiments, the alkyl group contains 1 to 3 carbon atoms, e.g., 1, 2, or 3 carbon atoms. Some non-limiting examples of alkyl include methyl, ethyl, propyl, 2-propyl (isopropyl), n-butyl, iso-butyl, sec-butyl, tert-butyl, and 2,2-dimethylpropyl. A particularly preferred, but non-limiting, example of alkyl is methyl.

[0014] The term "alkoxy" refers to an alkyl group, as defined above, attached to the parent molecular moiety through an oxygen atom. An alkoxy group contains 1 to 12 carbon atoms, unless otherwise specified. In some preferred embodiments, an alkoxy group contains 1 to 6 carbon atoms ("C1-C6-alkoxy"). In other embodiments, an alkoxy group contains 1 to 4 carbon atoms. In still other embodiments, an alkoxy group contains 1 to 3 carbon atoms. Some non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and tert-butoxy. A particularly preferred, but non-limiting, example of an alkoxy is methoxy.

[0015] The term "alkoxycarbonyl" refers to an alkoxy group, as defined above, attached to the parent molecular moiety through a carbonyl group. In some preferred embodiments, the alkoxy portion of the alkoxycarbonyl group contains 1 to 6 carbon atoms ("C1-C6-alkoxycarbonyl"). In other embodiments, the alkoxy group contains 1 to 4 carbon atoms. In still other embodiments, the alkoxy group contains 1 to 3 carbon atoms. Some non-limiting examples of alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl, isobutoxycarbonyl, and tert-butoxycarbonyl. A particularly preferred, but non-limiting, example of alkoxy is tert-butoxycarbonyl.

[0016] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of the free base or free acid, without being biologically or otherwise undesirable. Salts are formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, 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, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine, and the like. These salts may also be prepared by adding an inorganic or organic base to the 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 primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyimine resins, etc. A particular pharmaceutically acceptable salt of the compound of formula (I) is the hydrochloride salt.

[0017] The compounds of formula (I) may contain several asymmetric centers and may exist as optically pure enantiomers, mixtures of enantiomers, e.g. racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereomeric racemates or mixtures of diastereomeric racemates.

[0018] According to the Cahn-Ingold-Prelog rules, the asymmetric carbon atom may be of the "R" or "S" configuration.

[0019] The abbreviation "MAGL" refers to the enzyme monoacylglycerol lipase. The terms "MAGL" and "monoacylglycerol lipase" are used interchangeably herein.

[0020] Compounds of the Invention

[0021] In a first aspect, the present invention provides a compound of formula (I) or (II): [ka] [ka] (In the formula, R 1 teeth,: [ka] (wherein the wavy line indicates the R 1 (indicating the attachment point of Selected from; R 2 teeth,: [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1 or 2; V is selected from hydrogen and methyl; W is methyl, phenyl, [ka] (Wherein, Ar is C6-C 10 aryl or 5-14 membered heteroaryl) Selected from; X is selected from O, S, Si(CH3)2, and CH2; Y is selected from OH, NH, N(CH), N(CD, and azetidin-1-yl. Selected from; R 3 is selected from hydrogen, C1-C6-alkoxycarbonyl, and C1-C6-alkoxycarbonyl-NH-C1-C6-alkyl-NH-C(O)-; A is, (i) [ka] and; L is selected from -CH2-, -CH2O-, -OCH2-, -CH2OCH2-, and -O-; or (ii) [ka] (In the formula, (i) X is CH; L is selected from -CH2-, -CH2O-, -OCH2-, -CH2OCH2-, and -O-; or (ii) X and L together represent the group: [ka] wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I) or (II), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forming and; wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I) or (II); n is an integer selected from 1, 2, 3, and 4. or a pharmaceutically acceptable salt thereof.

[0022] In one embodiment, the present invention provides In formula (I), R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of Selected from; In formula (II), R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of Selected from; R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1 or 2; V is selected from hydrogen and methyl; W is methyl, phenyl, [ka] (Wherein, Ar is C6-C 10 aryl or 5-14 membered heteroaryl) Selected from; X is selected from O, S, Si(CH3)2, and CH2; Y is selected from OH, NH, N(CH), N(CD, and azetidin-1-yl. Selected from; R 3 is selected from hydrogen, C1-C6-alkoxycarbonyl, and C1-C6-alkoxycarbonyl-NH-C1-C6-alkyl-NH-C(O)-; A, (i) [ka] and; L is selected from -CH2-, -CH2O-, -OCH2-, -CH2OCH2-, and -O-; or (ii) [ka] (In the formula, (i) X is CH; L is selected from -CH2-, -CH2O-, -OCH2-, -CH2OCH2-, and -O-; or (ii) X and L together represent the group: [ka] wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I) or (II), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forming and; wherein the wavy line indicates the point of attachment of A and L, and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I) or (II); n is an integer selected from 1, 2, 3, and 4; Provided are compounds of formula (I) or (II) as described herein, or pharmaceutically acceptable salts thereof.

[0023] In one aspect, the present invention provides a compound of formula (I): [ka] (In the formula, R 1 , R 2 , A, L, and n are as defined herein. or a pharmaceutically acceptable salt thereof.

[0024] In one aspect, the present invention provides a compound of formula (II): [ka] (In the formula, R 1 , R 3 , A and L are as defined herein. or a pharmaceutically acceptable salt thereof.

[0025] In one embodiment, the present invention provides a compound comprising R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of The present invention provides a compound of formula (I) or (II) as described herein, selected from:

[0026] In one embodiment, the present invention provides a compound comprising R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of or a pharmaceutically acceptable salt thereof.

[0027] In one embodiment, the present invention provides a compound comprising R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of or a pharmaceutically acceptable salt thereof.

[0028] In one embodiment, A is (i) [ka] and; L is selected from -CH2O-, -OCH2-, -CH2OCH2-, and -O-; or (ii) [ka] (In the formula, (i) X is CH; L is selected from —CH— and —CHO—; or (ii) X and L together represent the group: [ka] wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forming and; wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I). Provided herein are compounds of formula (I), or pharmaceutically acceptable salts thereof.

[0029] In one embodiment, the present invention provides A, (i) [ka] and L is -CH2OCH2-; or (ii) [ka] (In the formula, (i) X is CH and L is -CH2-; or (ii) X and L together represent the group: [ka] wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forming and; wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I). Provided herein are compounds of formula (I), or pharmaceutically acceptable salts thereof.

[0030] In a preferred embodiment, the present invention comprises: A, (i) [ka] and L is -CH2OCH2-; or (ii) [ka] (wherein X is CH) and L is -CH2-; wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I). Provided herein are compounds of formula (I), or pharmaceutically acceptable salts thereof.

[0031] In certain preferred embodiments, the present invention provides A, [ka] and L is -CH2OCH2-, wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I). Provided herein are compounds of formula (I), or pharmaceutically acceptable salts thereof.

[0032] In certain preferred embodiments, the present invention provides a method for producing a pharmaceutical composition comprising: A, [ka] (wherein X is CH) and L is -CH2-; wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I). Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0033] In one embodiment, the present invention provides R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1; V is methyl; W is methyl; X is selected from O and Si(CH3)2; Y is N(CH3)2) Selected from; n is an integer selected from 1, 2, and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0034] In a preferred embodiment, the present invention comprises: R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1; V is methyl; W is methyl; X is selected from O and Si(CH3)2; Y is N(CH3)2) Selected from; n is an integer selected from 1, 2, and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0035] In a preferred embodiment, the present invention comprises: R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1; V is methyl; W is methyl; X is selected from O and Si(CH3)2; Y is N(CH3)2) Selected from; n is an integer selected from 1 and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0036] In a particularly preferred embodiment, the present invention comprises: R 2 but, [ka] (wherein the wavy line represents the R 2 (indicating the attachment point of and; n is an integer selected from 1 and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0037] In a particularly preferred embodiment, the present invention comprises: R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; V is methyl; W is methyl) and; n is an integer selected from 1 and 4; Provided herein are compounds of formula (I), or pharmaceutically acceptable salts thereof.

[0038] In a particularly preferred embodiment, the present invention comprises: R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; X is selected from O and Si(CH3)2; Y is N(CH3)2) and; n is an integer selected from 1 and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0039] In a particularly preferred embodiment, the present invention comprises: R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1) and; n is an integer selected from 1 and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0040] In one embodiment, the present invention provides A, (i) [ka] and L is selected from -OCH2-, -CH2OCH2-, and -O-; or (ii) [ka] (wherein X is CH) and L is -CH2O-; where the wavy line indicates the point of attachment to L and the asterisk indicates the point of attachment to the carbonyl group of formula (II). Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0041] In one embodiment, the present invention provides R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of Selected from; R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1; V is methyl; W is methyl; X is selected from O and Si(CH3)2; Y is N(CH3)2) Selected from; A, (i) [ka] and; L is selected from -CH2O-, -OCH2-, -CH2OCH2-, and -O-; or (ii) [ka] (In the formula, (i) X is CH; L is selected from —CH— and —CHO—; or (ii) X and L together represent the group: [ka] wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forming and; where the wavy line indicates the point of attachment to L and the asterisk indicates the point of attachment to the carbonyl group of formula (I); n is an integer selected from 1, 2, and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0042] In a preferred embodiment, the present invention comprises: R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of Selected from; R 2 but, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1; V is methyl; W is methyl; X is selected from O and Si(CH3)2; Y is N(CH3)2) Selected from; A, (i) [ka] and L is -CH2OCH2-; or (ii) [ka] (In the formula, (i) X is CH and L is -CH2-; or (ii) X and L together represent the group: [ka] wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forming and; where the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I); n is an integer selected from 1, 2, and 4; Provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0043] In a particularly preferred embodiment, the present invention comprises: R 1 but, [ka] (wherein the wavy line represents the R 1 (indicating the attachment point of Selected from; R 2 teeth, [ka] (In the formula, The wavy line represents the R 2 indicates the point of attachment; p is 1; V is methyl; W is methyl; X is selected from O and Si(CH3)2; Y is N(CH3)2) Selected from; A, (i) [ka] and L is -CH2OCH2-; or (ii) [ka] (wherein X is CH) and L is -CH2-; where the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I); n is an integer selected from 1 and 4; Provided herein are compounds of formula (I), or pharmaceutically acceptable salts thereof.

[0044] In one embodiment, the present invention provides benzyl(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.5]nonan-7-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(3-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamate; benzyl(6-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)hexyl)carbamate; benzyl(2-(2-(((1-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; Cyclohex-1-en-1-ylmethyl(2-(2-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((7-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-7-azaspiro[3.5]nonan-2-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)pentyl)carbamate; benzyl(2-(2-((((1R,5S)-3-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl (R)-(2-(2-(((1-(3-(2-oxooxazolidin-4-yl)propanoyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((1-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; 2-(6-(2-(2-((7-nitro-1l2,3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one; (4aR,8aS)-6-(3-(((2-(2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; 2-(3-(((2-(2-((7-nitro-1H-3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one; (4aR,8aS)-6-(3-(((2-(3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)propoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(3-(((2-((5-((7-nitro-1,3-dihydrobenzo[c][1,2,5]oxadiazol-4-yl)amino)pentyl)oxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(2-(2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(3-(2-((7-nitro-1,3-dihydrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; 2-(7-(dimethylamino)-3-(dimethyliminio)-5,5-dimethyl-3,5-dihydrodibenzo[b,e]silin-10-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 3,3-dimethyl-1-(6-oxo-6-((2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium 2,2,2-trifluoroacetate; 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborin-3-yl)-N-(2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)propanamide; 2,2,2-Trifluoroacetic acid, 3,3-dimethyl-1-(6-oxo-6-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium salt; 3,3-dimethyl-1-(6-oxo-6-((2-(2-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium 2,2,2-trifluoroacetate; 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborin-3-yl)-N-(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)propanamide; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate compounds and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((5-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)pentyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)pentyl)carbamoyl)benzoate; Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate; Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoate; Ethyl 2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate; tert-Butyl (2-(3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzamido)ethyl)carbamate; tert-Butyl (2-(2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzamido)ethyl)carbamate; (4aR,8aS)-6-(3-((benzyloxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; and (4aR,8aS)-6-(3-(phenoxymethyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one The present invention provides a compound of formula (I) or (II) as described herein, selected from:

[0045] In one embodiment, the present invention provides benzyl(6-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)hexyl)carbamate; benzyl(2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)pentyl)carbamate; benzyl (R)-(2-(2-(((1-(3-(2-oxooxazolidin-4-yl)propanoyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((1-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; 2-(6-(2-(2-((7-nitro-1l2,3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one; (4aR,8aS)-6-(3-(((2-(2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; 2-(3-(((2-(2-((7-nitro-1H-3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one; (4aR,8aS)-6-(3-(((2-(3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)propoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(3-(((2-((5-((7-nitro-1,3-dihydrobenzo[c][1,2,5]oxadiazol-4-yl)amino)pentyl)oxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(2-(2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(3-(2-((7-nitro-1,3-dihydrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; 2-(7-(dimethylamino)-3-(dimethyliminio)-5,5-dimethyl-3,5-dihydrodibenzo[b,e]silin-10-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 3,3-dimethyl-1-(6-oxo-6-((2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium 2,2,2-trifluoroacetate; 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborin-3-yl)-N-(2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)propanamide; 2,2,2-Trifluoroacetic acid, 3,3-dimethyl-1-(6-oxo-6-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium salt; 3,3-dimethyl-1-(6-oxo-6-((2-(2-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium 2,2,2-trifluoroacetate; 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborin-3-yl)-N-(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)propanamide; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate compounds and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((5-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)pentyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)pentyl)carbamoyl)benzoate The present invention provides a compound of formula (I) as described herein, selected from:

[0046] In one embodiment, the present invention provides benzyl(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.5]nonan-7-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(3-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamate; benzyl(2-(2-(((1-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; Cyclohex-1-en-1-ylmethyl(2-(2-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((7-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-7-azaspiro[3.5]nonan-2-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-((((1R,5S)-3-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)methoxy)methyl)phenoxy)ethyl)carbamate; Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate; Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoate; Ethyl 2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate; tert-Butyl (2-(3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzamido)ethyl)carbamate; tert-Butyl (2-(2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzamido)ethyl)carbamate; (4aR,8aS)-6-(3-((benzyloxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; and (4aR,8aS)-6-(3-(phenoxymethyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one The present invention provides a compound of formula (II) as described herein, selected from:

[0047] In certain embodiments, the present invention provides pharmaceutically acceptable salts of compounds according to formula (I) described herein. In more particular embodiments, the present invention provides compounds according to formula (I) described herein as the free base.

[0048] Manufacturing method The preparation of compounds of formula (I) or (II) of the present invention may be carried out by sequential or convergent synthetic routes. The synthesis of the present invention is illustrated in the following general scheme. The skills required to carry out the reactions and purification of the resulting products are known to those skilled in the art. The substituents and indices used in the following process descriptions have the meanings indicated herein unless otherwise indicated.

[0049] If one of the starting materials, intermediates, or compounds of formula (I') or (I) contains one or more functional groups that are not stable or reactive under the reaction conditions of one or more reaction steps, suitable protecting groups (such as those described in "Protective Groups in Organic Chemistry," by T.W. Greene and P.G.M. Hutts, 5th Edition, 2014, John Wiley & Sons, NY) may be introduced prior to a critical step by applying methods well known in the art. Such protecting groups may be removed at a later stage of the synthesis using standard methods described in the literature.

[0050] When the starting materials or intermediates contain a stereocenter, the compounds of formula (I) or (II) are obtained as mixtures of diastereomers or enantiomers, which can be separated by methods well known in the art, such as chiral HPLC, chiral SFC, or chiral crystallization. Racemates can be separated into their antipodes via diastereomeric salts, for example, by crystallization with optically pure acids, or by separating the antipodes by specific chromatographic methods using either chiral adsorbents or chiral eluents. Starting materials and intermediates containing a stereocenter can also be separated to obtain diastereomerically / enantiomerically enriched starting materials and intermediates. The use of such diastereomerically / enantiomerically enriched starting materials and intermediates in the synthesis of compounds of formula (I) or (II) generally results in the respective diastereomerically / enantiomerically enriched compounds of formula (I) or (II).

[0051] Those skilled in the art will recognize that in the synthesis of compounds of formula (I) or (II), unless otherwise desired, an "orthogonal protecting group strategy" may be applied to cleave some protecting groups one 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 described 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).

[0052] Those skilled in the art will recognize that the reaction sequence may vary depending on the reactivity and nature of the intermediates.

[0053] More specifically, compounds of formula (I) or (II) can be prepared by the methods described below, those described in the Examples, or similar methods. Suitable reaction conditions for the individual reaction steps are known to those skilled in the art. For literature-described reaction conditions affecting the described reactions, 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 reactions could be easily carried out with or without a solvent. There is no particular restriction on the nature of the solvent used, so long as it does not adversely affect the reaction or the reagents involved and is capable of dissolving the reagents to at least some extent. The described reactions can occur over a wide range of temperatures, and the exact reaction temperature is not critical to the present invention. The described reactions are conveniently carried out at temperatures ranging from -78°C to reflux. The time required for the reaction can also vary widely, depending on many factors, particularly the reaction temperature and the nature of the reagents. However, a period of 0.5 hours to several days is usually sufficient to obtain the intermediates and compounds described. The reaction order is not limited to the order shown in the scheme, but the order of the reaction steps can be freely changed depending on the starting materials and their respective reactivities.

[0054] If the starting materials or intermediates are not commercially available or their synthesis is not described in the literature, they can be prepared analogously to existing procedures for similar analogs or as outlined in the experimental section.

[0055] The following abbreviations are used herein: AcOH = acetic acid, ACN = acetonitrile, Boc = tert-butyloxycarbonyl, CAS RN = Chemical Abstracts Registry Number, Cbz = benzyloxycarbonyl, CsCO 3=Cesium carbonate, CO = carbon monoxide, DMAP = 4-dimethylaminopyridine, DMF = N,N-dimethylformamide, DMP = Dess-Martin periodinane, DIBAL-H = diisobutylaluminum hydride, DIPEA = N,N-diisopropylethylamine, dppf = 1,1-bis(diphenylphosphino)ferrocene, EDC.HCl = N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, ESI = electrospray ionization, EtOAc = ethyl acetate, EtOH = ethanol, h = time (s), FA = formic acid, H2O = water, H2SO4 = sulfuric acid, Hal = halogen, HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate, HBTU = O-benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluorophosphate ate, HCl = hydrogen chloride, HOBt = 1-hydroxy-1H-benzotriazole; HPLC = high performance liquid chromatography, K2CO3 = potassium carbonate, KHCO3 = potassium bicarbonate, KI = potassium iodide, KOH = potassium hydroxide, K3PO4 = tripotassium phosphate, KOtBu = potassium tert-butoxide, LiAlH4 or LAH = lithium aluminum hydride, LiHMDS = lithium bis(trimethylsilyl)amide, LiOH = lithium hydroxide, LiTMP = lithium tetramethyl-piperidine, MgSO4 = Magnesium sulfate, min = minute, mL = milliliter, MPLC = medium pressure liquid chromatography, MS = mass spectrum, Ms = mesyl, NaH = sodium hydride, NaHCO3 = sodium bicarbonate, NaOH = sodium hydroxide, Na2CO3 = sodium carbonate, Na2SO4 = sodium sulfate, Na2S2O3 = sodium thiosulfate, NBS = N-bromosuccinimide, nBuLi = n-butyllithium, NEt3 = triethylamine (TEA), NH4Cl = ammonium chloride, NMP = N-methyl-2-pyrrolidone, OA c = acetoxy, T3P = propylphosphonic anhydride, PG = protecting group, Pd-C = palladium on activated carbon, Pd(dppf)Cl2 = 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride, Pd2(dba)3 = tris(dibenzylideneacetone)dipalladium(0), Pd(OAc)2 = palladium(II) acetate, Pd(OH)2 = palladium hydroxide, Pd(PPh3)4 = tetrakis(triphenylphosphine)palladium(0), PTSA = p-toluenesulfonic acid, Ts, tosyl, R = any group rt= room temperature, SFC = supercritical fluid chromatography, TBAI = tetrabutylammonium iodide, TEA = triethylamine, TFA = trifluoroacetic acid, THF = tetrahydrofuran, TMEDA = N,N,N',N'-tetramethylethylenediamine.

[0056] A, L, n, R 1 and R 2 Compounds of formula I, wherein: is as described herein, may be synthesized analogously to literature procedures and / or as shown, for example, in Scheme 1. [ka]

[0057] Scheme 1 Thus, amine derivative 1 has a suitable nitrogen protecting group "PG" such as benzyloxycarbonyl (Cbz) carbamate at the end of the alkane chain and an orthogonal nitrogen protecting group "PG" attached to the cyclic amine such as tert-butyloxycarbonyl (Boc) carbamate. 1 The protecting group PG can be obtained by applying a method known in the art (e.g., a Boc group using TFA in DCM at a temperature between 0° C. and room temperature, a Cbz group using hydrogen on activated carbon in a suitable solvent such as MeOH, EtOH, EtOAc or a mixture thereof in the presence of a suitable catalyst such as Pd or Pd(OH) 2 , as described in, for example, "Protective Groups in Organic Chemistry" by T.W. Greene and P.G.Wuts, 4th Ed., 2006, Wiley NY). 1 Intermediate 1 is subsequently removed, and the resulting free amine is converted to a compound of type 2a (R 2a containing an N at the point of attachment to the remainder of formula I) in the presence of a urea-forming reagent, such as bis(trichloromethyl) carbonate, using a suitable base and solvent, such as sodium bicarbonate in DCM. 1The reaction with the carbamoyl chloride (C moiety) gives compound 3, which contains the compound of formula Ia (Step a1). Additional urea-forming reagents include, but are not limited to, phosgene, trichloromethyl chloroformate, (4-nitrophenyl) carbonate, or 1,1'-carbonyldiimidazole. This type of reaction and the use of these reagents have been widely described in the literature (e.g., G. Sartori et al., Green Chemistry 2000, 2, 140). Those skilled in the art will recognize that the order of addition of reagents can be important in this type of reaction due to the reactivity and stability of the carbamoyl chloride formed as an intermediate, as well as to avoid the formation of undesired symmetrical urea by-products. Alternatively, R containing a C at the point of attachment to the remainder of formula I can be used. 1 The moiety was introduced as a carboxylic acid, which was then converted into PG 1 The intermediate amine obtained after removal is coupled to form the respective amide containing compound of formula Ia (step a2). This type of amide coupling can be achieved by using one of the well-known coupling reagents such as DCC, HATU, EDCI, HOBt, TBTU, T3P, and a base such as Hunig's base, triethylamine, or DMAP in a suitable solvent such as N,N-dimethylformamide, DMA, DCM, or 1-4-dioxane, preferably at 0° C. to room temperature.

[0058] Removal of the protecting groups in intermediate 3 applying methods known in the art (e.g., Boc group using TFA in DCM at temperatures between 0° C. and room temperature, Cbz group using hydrogen in the presence of a suitable catalyst such as Pd on charcoal or Pd(OH) in a suitable solvent such as MeOH, EtOH, EtOAc, or a mixture thereof, and groups as described, for example, in Protective Groups in Organic Chemistry, T.W. Greene and P.G.M. Buts, 4th Edition, 2006, Wiley, NY) gives intermediate 4 (step b).

[0059] Reaction of amine 4 with a fluorescent label containing a carboxylic acid moiety affords compounds of formula Ib (R 2= fluorescent label) is obtained (step c). This type of amide coupling can be achieved by using one of the well-known coupling reagents, such as DCC, HATU, EDCI, HOBt, TBTU, or T3P, and a base, such as Huenig's base, triethylamine, or DMAP, in a suitable solvent, such as N,N-dimethylformamide, DMA, DCM, or dioxane, preferably at 0°C to room temperature. Alternatively, a fluorescent label containing an N-hydroxysuccinimide (NHS) activated ester is reacted with 4 in the presence of a suitable base, such as N,N-diisopropylethylamine (DIPEA), 2,6-lutidine, or the like, in a suitable solvent, such as DCM, THF, ACN, DMF, or dioxane, preferably at 0°C to room temperature, to give compounds of formula Ib. To form the final molecule Ib bearing the nitrobenzofurazan dye, amine 4 can be reacted in a nucleophilic aromatic substitution, for example, with 4-chloro-7-nitrobenzene-2-oxa-1,3-diazole (CAS RN10199-89-0) or 4-fluoro-7-nitrobenzene-2-oxa-1,3-diazole (CAS RN29270-56-2) in a solvent such as MeOH, preferably at 0° C. to room temperature, in the presence of a base such as triethylamine (step c). Optionally, the amine derivative 1, in which "PG" is a fluorescent label, can be directly converted to a compound of formula Ib according to step a.

[0060] Protected amines 1 and their amine 2a or carboxylic acid 2b counterparts are either commercially available or can be prepared according to literature methods or as shown below.

[0061] Intermediates of type 1, such as intermediates 1a and 1b, can be prepared by a variety of conditions, which can be exemplified by the general synthetic procedure outlined in Scheme 2. [ka]

[0062] Scheme 2 Intermediates 5, 6, and 11 are commercially available or can be prepared according to literature methods. The functional hydroxyl groups of 5 and 11 can be utilized to carry out nucleophilic substitution (S) in the presence of a suitable base (e.g., CsCO, KCO) in a suitable solvent (e.g., DMF) at temperatures preferably between room temperature and 80°C. N Alkane chain derivatives of type 6 can be attached via a 2-methyl-2-propanol (2-methyl-2-propanol) reaction to give compounds 7 and 12, respectively (step d). Here, "LG" is a suitable leaving group, such as, but not limited to, methanesulfonate (OM), p-toluenesulfonate (OT), bromine, iodide, etc. Intermediates of type 7 and 12 can also be obtained by literature methods or by other etherification reactions under Mitsunobu conditions, for example, using PPh3 and diethyl azodicarboxylate. Benzoates of type 7 can be converted to their corresponding benzyl alcohols 8 by treatment with a suitable reducing agent such as LiAlH, LiBH, diisobutylaluminum hydride (DIBAL-H) ​​in a suitable solvent such as toluene at temperatures preferably between −78° C. and 0° C. (step e).

[0063] Benzyl alcohols 8 can be converted to their benzyl bromide analogues 9 by reactions described in the literature, such as the Appel reaction with CBr4 and PPh3 or by the use of PBr3 in a suitable solvent, such as DCM, preferably at temperatures between 0 and 25 °C (step f).

[0064] Molecules of type 1a can be obtained by reacting benzyl bromide 9 with alcohol 10 in a nucleophilic substitution reaction in a suitable solvent such as THF in the presence of a suitable base, such as KOtBu, at temperatures preferably between room temperature and 80° C. (step g). Those skilled in the art will recognize that other etherification methods described in the literature can be used to prepare 1a, as well as analogs of bromine 9 bearing other suitable leaving groups in place of the bromine (e.g., I, OM, OT).

[0065] Boronic acids, esters, and their derivatives are known as versatile building blocks for transition metal-mediated cross-coupling reactions. Intermediates of type 1b can be readily obtained via cross-coupling with a suitable building block, such as vinyl boronate ester 13, in the presence of a suitable base, such as KCO, a suitable water-containing solvent mixture, such as water / dioxane, and a transition metal catalyst, such as, but not limited to, Pd(dppf)Cl, preferably at temperatures between 25 and 100 °C (step h). Alternatively, hydrolysis of vinyl boronate 13 by methods known to those skilled in the art, such as treatment with an ammonium acetate-buffered water / acetone mixture and sodium periodate, can be carried out as a separate step, followed by (step h) in the absence of water.

[0066] Vinylboronic acid ester 13 is commercially available or can be prepared according to literature methods or as shown below. Vinylboronates 13 (e.g., vinylboronic acid pinacol ester) can be readily obtained by treating ketone 14 with bis((pinacolato)boryl)methane (CAS RN 78782-17-9) in the presence of a suitable base, such as lithium tetramethylpiperidine (LiTMP), as described in the literature (Kovalenko et al., The Boron-Wittig Olefination of Aldehydes and Ketones with Bis[(pinacolato)boryl]methane: an Extended Reaction Scope. Eur J. Org. Chem., 2019:5264-5635). The general synthetic method is shown in Scheme 3 (Step i). [ka]

[0067] Scheme 3 A, L, R 1 and R 3 Compounds of formula II, wherein: is as described herein, can be synthesized analogously to literature procedures and / or as shown, for example, in Scheme 4. [ka]

[0068] Scheme 4 The protecting group "PG" as described for Intermediate 1 1 " and amines of type 15 bearing leaving group "LG" as described for intermediate 6 are both commercially available and can be easily obtained in one step from alcohol derivative 10 by methods widely described in the literature (reaction with p-toluenesulfonic acid chloride in the presence of DMAP and TEA, or Comprehensive Organic Transformations: A Guide to Functional Group Preparations, 2nd Edition, Richard C. Larock. John Wiley & Sons, New York, NY. 1999, step j).

[0069] Phenolic ester derivative 16 can be reacted with intermediate 15 via nucleophilic substitution to give ether intermediate 17 (step d), similar to the method described for the conversion of 5 to 7 in Scheme 2. Compounds of type IIa can be obtained from 17 similarly to the conversion of 1 to 3 in Scheme 1 (step a).

[0070] Compounds of formula IIb can be obtained, for example, from the reaction of IIa with amine 18 under amide coupling conditions, for example by using one of the well-known coupling reagents such as DCC, HATU, EDCI, HOBt, TBTU, T3P, and a base such as Huenig's base, triethylamine or DMAP in a suitable solvent such as N,N-dimethylformamide, DMA, DCM or dioxane, preferably at room temperature (step k). Treatment of benzyl bromide 19 with alcohols of type 10 (see above) in the presence of a suitable base, such as sodium hydride (NaH), and a suitable solvent, such as a DMF / DCM mixture, under nucleophilic substitution conditions, preferably at temperatures between 0°C and 25°C, affords ether intermediate 20 (step 1). Compounds having formula IIc are those having R in the remainder of formula II. 3 By applying =H, it is readily available from 20, similar to that described for the conversion of 1 to 3 in Scheme 1 (step a).

[0071] In one aspect, the present invention provides a method for preparing a compound of formula (I) described herein, comprising the step of: [ka] (In the formula, A, L, R 1 and n is as defined herein. of, (a) with a fluorescent label containing a carboxylic acid moiety in the presence of a coupling reagent and a base; or (b) with a fluorescent label containing an N-hydroxysuccinimide (NHS) activated ester in the presence of a base; or (c) with 4-chloro-7-nitrobenz-2-oxa-1,3-diazole or 4-fluoro-7-nitrobenz-2-oxa-1,3-diazole in the presence of a base reacting to obtain a compound of formula (I).

[0072] In one embodiment, the fluorescent label comprising a carboxylic acid moiety in reaction (a) is [ka] wherein X, Y, V, W, and p are as defined herein. is selected from.

[0073] In one embodiment, the amide coupling reagent is selected from DCC, HATU, EDCI, HOBt, TBTU, and T3P.

[0074] In one embodiment, the base in reaction (a) is selected from Hunig's base, trimethylamine, and DMAP.

[0075] In one embodiment, reaction (a) is carried out in a suitable solvent, preferably a solvent selected from N,N-dimethylformamide, DMA, DCM and dioxane.

[0076] In one embodiment, the fluorescent label containing an N-hydroxysuccinimide (NHS) activated ester in reaction (b) is [ka] wherein X, Y, V, W, and p are as defined herein. is selected from.

[0077] In one embodiment, the base in reaction (b) is selected from N,N-diisopropylethylamine (DIPEA) and 2,6-lutidine.

[0078] In one embodiment, the reaction (b) is carried out in a suitable solvent, preferably a solvent selected from DCM, THF, ACN, DMF and dioxane.

[0079] In one embodiment, the base in reaction (c) is triethylamine.

[0080] In one embodiment, the reaction (c) is carried out in a suitable solvent, preferably methanol.

[0081] In one aspect, the present invention provides a compound of formula (I) or (II) as described herein when prepared according to any one of the methods described herein.

[0082] MAGL inhibitory activity Compounds were profiled for MAGL inhibitory activity by measuring enzymatic activity via the hydrolysis of the natural substrate, 2-arachidonoylglycerol (2-AG), to produce arachidonic acid, which was followed by mass spectrometry (hereafter referred to as the "2-AG assay").

[0083] The 2-AG assay was performed in a 384-well assay plate (PP, Greiner, catalog no. 784201) in a total volume of 20 μL. Compound dilutions were performed in 3-fold dilutions in 100% DMSO (VWR Chemicals 23500.297) in polypropylene plates, resulting in a final assay concentration range of 12.5 μM to 0.8 pM. 0.25 μL of compound dilution (100% DMSO) was added to 9 μL of MAGL in 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 room temperature for 15 min. To initiate the reaction, 10 μL of 2-arachidonoylglycerol in assay buffer was added. The final concentrations in the assay were 50 pM MAGL and 8 μM 2-arachidonoylglyerol. After shaking and incubation at room temperature for 30 min, the reaction was quenched by adding 40 μL of ACN containing 4 μM d8-arachidonic acid. The amount of arachidonic acid was monitored using an online SPE system (Agilent Rapidfire) coupled to a triple quadrupole mass spectrometer (Agilent 6460). A C18 SPE cartridge (G9205A) was used with an ACN / water liquid setup. The mass spectrometer was operated in negative electrospray mode, following the mass transitions of arachidonic acid (303.1 → 259.1) and d8-arachidonic acid (311.1 → 267.0). Compound activity was calculated based on the intensity ratio [arachidonic acid / d8-arachidonic acid].

[0084] [Table 1]

[0085] Uses of the Compounds of the Invention The compounds of formula (I) are fluorescent imaging probes with high affinity for MAGL. Therefore, they can be used as high-resolution tools to investigate the localization of MAGL in live cells, including its expression level and protein distribution, structure, dynamics, and function in health and disease. They can also be applied to cellular trafficking studies, for example, using flow cytometry (fluorescence-activated cell sorting) experiments or confocal live-cell imaging.

[0086] In one aspect, the present invention provides a compound of formula (I) as described herein for use in a monoacylglycerol lipase (MAGL) occupancy assay.

[0087] In a further aspect, the present invention provides a compound of formula (I) as described herein for use in diagnostic imaging of monoacylglycerol lipase (MAGL) in a mammal.

[0088] In a further aspect, the present invention provides a compound of formula (I) as described herein for use in generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

[0089] In a further aspect, the present invention provides the use of a compound of formula (I) as described herein in a monoacylglycerol lipase (MAGL) occupancy assay.

[0090] In a further aspect, the present invention provides the use of a compound of formula (I) as described herein in the diagnostic imaging of monoacylglycerol lipase (MAGL) in a mammal.

[0091] In a further aspect, the present invention provides the use of a compound of formula (I) as described herein for the generation of equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

[0092] In a further aspect, the present invention provides a method of testing monoacylglycerol lipase (MAGL) occupancy, comprising contacting MAGL with a compound of formula (I) described herein.

[0093] In a further aspect, the present invention provides a method for diagnostic imaging of monoacylglycerol lipase (MAGL) in a mammal, comprising contacting the MAGL with a compound of formula (I) described herein.

[0094] In a further aspect, the present invention provides a method for generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL), comprising contacting MAGL with a compound of formula (I) described herein.

[0095] The compound of formula (II) is a useful synthetic intermediate for preparing the fluorescent probe of formula (I). Thus, in one aspect, the present invention provides the use of a compound of formula (II) described herein for the preparation of the fluorescent probe of formula (I) described herein. [Example]

[0096] The present invention will be more fully understood by reference to the following examples, which should not, however, be construed as limiting the scope of the claims to the examples.

[0097] Where preparations are obtained as mixtures of enantiomers, the pure enantiomers may 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.

[0098] Unless otherwise stated, all reactions and intermediates were prepared under an argon atmosphere.

[0099] Example 1 Benzyl (2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate [ka] Step d Ethyl 2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzoate [ka] Ethyl salicylate (CAS RN118-61-6, 2.26 g, 13.6 mmol) was dissolved in DMF (8 mL), CsCO (4.43 g, 13.6 mmol) was added, and the mixture was stirred at room temperature for 5 min. Next, 2-(((benzyloxy)carbonyl)amino)ethyl 4-methylbenzenesulfonate (CAS RN93407-96-6, 2.80 g, 8.00 mmol) was added, and the mixture was stirred at 55 °C for 18 h. The reaction was quenched by diluting with DCM (60 mL), extracted with sn. 10% NaOH (2 × 40 mL), and washed with brine (50 mL). The organic layer was dried over MgSO, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel flash chromatography (0–50% AcOEt in cyclohexane) to give the title compound (2.54 g, 7.40 mmol, 92%). 1 H NMR(300MHz,CDCl3)δ 7.82(dd,J=7.8,1.8Hz,1H),7.45(ddd,J=8.2,7.4,1.8Hz,1H),7.41-7.24(m,5H),7.07-6.90(m,2H),5.98(s,1H,NHCbz),5.12(s ,2H,OCH2),4.35(q,J=7.1Hz,2H,OCH2CH3),4.14(t,J=5.0Hz,2H,OCH2),3.64(m,2H,CH2NHCbz),1.36(t,J=7.1Hz,3H,OCH2CH3). 13C NMR(75MHz,CDCl3)δ 166.2,158.44,156.73(C=O CO2Et,C=O Cbz,C Ar -O)136.8,133.7,131.9,128.6,128.11,128.09,121.1,120.9,114.3,68.8(OCH2),66.8(OCH2),61.1(OCH2),40.6(CH2NHCbz),14.4(CH3).C 19 H 21 NO5Na[M+Na] + Calculated for LC-HRMS (ESI): 366.1312; Found: 366.1301

[0100] Step e Benzyl (2-(2-(hydroxymethyl)phenoxy)ethyl)carbamate [ka] Ethyl 2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzoate (1.15 g, 3.35 mmol) was dissolved in toluene (11.2 mL) at 0 °C under a N atmosphere, and a 1 M solution of DIBAL in DCM (15.1 mL, 15.1 mmol) was added dropwise. The reaction mixture was stirred at 0 °C until completion (1.5 h, TLC monitoring). The reaction was diluted with DCM (20 mL) and quenched by the dropwise addition of AcOH (0.86 mL, 15.1 mmol). The mixture was then extracted with 10% aqueous sn. NaOH (2 × 40 mL) and washed with brine (40 mL). The organic layer was dried over MgSO, filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel flash chromatography (0 to 65% AcOEt in cyclohexane) to give the title compound (817 mg, 2.71 mmol, 81%). 1H NMR(300MHz,CDCl3)δ 7.39-7.20(m,4H),6.95(td,J=7.5,1.1Hz,1H),6.85(d,J=8.1Hz,1H),5.12(s,2H,OC H2),4.66(s,2H,OCH2),4.10(t,J=5.0Hz,2H,OCH2),3.63(t,J=5.0Hz,2H,CH2NHCbz). 13 C NMR(75MHz,CDCl3)δ 156.9,156.7(C=O Cbz,C Ar -O),136.5,129.5,129.4,129.3,128.7,128.3,121.3,111.6,67.6(OCH2),67.1(OCH2),62.1(OCH2),40.8(CH2-NHCbz).C 17 H 20 NO4[M+H] + Calculated for LC-HRMS (ESI): 302.1386; Found: 302.1365

[0101] Step f Benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate [ka] The alcohol benzyl (2-(2-(hydroxymethyl)phenoxy)ethyl)carbamate (795 mg, 2.64 mmol) was dissolved in DCM (20.3 mL) and cooled to 0°C. CBr4 (1.00 g, 3.04 mmol) was then added, followed by PPh3 (796 mg, 3.04 mmol, 1.5 M solution in DCM). The reaction mixture was stirred at room temperature until completion (10 min, TLC monitoring). The reaction was quenched by diluting with DCM (30 mL), adding saturated NaHCO3 (2 × 30 mL), and washing with brine (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel flash chromatography (0–40% AcOEt in cyclohexane) to give the title compound (694 mg, 1.90 mmol, 72%). 1H NMR(300MHz,CDCl3)δ 7.38-7.28(m,7H),7.04-6.81(m,2H),5.12(s,2H,OCH2Cbz),4.59(d,J=27.9Hz,2H,CH 2-Br),4.14(t,J=4.9Hz,2H,OCH2CH2-NHCbz),3.68(q,J=5.3Hz,2H,OCH2CH2-NHCbz). 13 C NMR(75MHz,CDCl3)δ 156.7,136.7,131.0,130.6,128.7,128.1,121.2,111.8,67.5(-OCH2-),66.9 (-OCH2-),40.7(-OCH2CH2-NHCbz),29.7(CH2-Br).LC-MS(ESI):386.0[M+Na] + .

[0102] Step g tert-Butyl 3-(((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carboxylate (BB1) [ka] tert-Butyl 3-(hydroxymethyl)azetidine-1-carboxylate (CAS RN142253-56-3, 196 mg, 1.05 mmol) was dissolved in anhydrous THF (7 mL) at 0 °C, KOtBu (118 mg, 1.05 mmol) was added, and the mixture was stirred for 5 min. The bromine derivative benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate (254.9 mg, 0.70 mmol) was then dissolved in anhydrous THF (1.5 mL) and added dropwise to the reaction mixture. The mixture was then warmed to room temperature and stirred for 4 h until completion (TLC monitoring). The reaction was diluted with DCM (10 mL) and quenched by the dropwise addition of AcOH (0.1 mL, 1.05 mmol). The mixture was extracted with saturated aqueous NaHCO3 (2 × 20 mL) and washed with brine (20 mL). The organic layer was dried over MgSO, filtered, and concentrated under reduced pressure, and the concentrate was purified by silica gel flash chromatography (0-40% AcOEt in cyclohexane) to give the title compound (186 mg, 0.40 mmol, 57%). 1 H NMR(300MHz,CDCl3)δ 7.39-7.23(m,7H),6.96(td,J=7.4,1.0Hz,1H),6.85(d,J=8.2Hz,1H),5.50(s,1H,NHCbz),5.10(s,2H,OCH 2),4.54(s,2H,OCH2),4.14-4.03(m,2H,OCH2),3.86(t,J=8.4Hz,2H,OCH2),3.67-3.51(m,6H,CH2NHCbz,2 x N-CH2Azetidine),2.78-2.63(m,1H,CHAzetidine),1.43(s,9H,3 x CH3Boc).13C NMR(75MHz,CDCl3)δ 156.5,136.5,129.9,129.4,128.7,128.44,128.38,126.7,121.2,112.1,79.4(C(CH3)Boc) ,72.7(CH azetidine),68.8(OCH2),67.6(OCH2),67.0(OCH2),40.8(CH2-NHCbz),28.5(CH3Boc).C 26 H 35 N2O6[M+H] +Calculated for LC-HRMS (ESI): 471.2490; Found: 471.2475.

[0103] Step a1 Benzyl (2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate 1. Boc Deprotection: The N-Boc protected derivative tert-butyl 3-(((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carboxylate (BB1, 93.0 mg, 0.20 mmol) was dissolved in DCM (1.3 mL) at 0 °C, and TFA (152 μL, 2.0 mmol) was added. The reaction mixture was stirred at that temperature until Boc deprotection was complete (1 h, LC-MS monitoring). The reaction mixture was diluted with toluene (3.00 mL) and coevaporated twice with toluene (3.00 mL) under reduced pressure. The 2,2,2-trifluoroacetate salt of the free amine 3-(((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine was used in the next step (2) without further purification. 2. Urea Coupling: (4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazin-6-ium (2S,3S)-3-carboxy-2,3-bis((4-methylbenzoyl)oxy)propanoate salt (CAS RN 2624363-49-9, 119 mg, 0.16 mmol) was dissolved in anhydrous acetonitrile (0.7 mL), and 1,1'-carbonyl-di-(1,2,4-triazole) (CAS RN 41864-22-6, 26.3 mg, 0.16 mmol) was added, followed by triethylamine (0.16 mL, 1.12 mmol). The reaction mixture was stirred at room temperature for 2 h. The 2,2,2-trifluoroacetate salt of the amine nucleophile from Step 1, 3-(((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine, was then dissolved in ACN (0.5 mL) and added dropwise to the reaction mixture. The reaction mixture was heated to 50 °C and stirred at that temperature for 2.5 h. The reaction was quenched by diluting with DCM (5.00 mL), adding saturated aqueous NaHCO (2.00 mL), and washing with brine (2.00 mL). The organic layer was dried over MgSO, filtered, and concentrated under reduced pressure. The concentrate was purified by HPLC (25-75% ACN in HO with 0.1% TFA) to give the title compound (70.0 mg, 0.13 mmol, 79%). 1 H NMR(300MHz,MeOD)δ 7.37-7.16(m,7H,H-Ar),6.96-6.84(m,2H,H-Ar),5.05(s,2H,O-CH2-),4.51(s,2H,O-CH2-),4.25-4.06(m,2H),4.06-3.88(m,4H),3.8 2-3.62(m,3H),3.58-3.45(m,4H),3.32-3.20(m,3H),2.99-2.83(m,2H),2.80-2.66(m,1H,-CH-azetidine),1.90-1.64(m,2H.-CH2-HHPO). 13 C NMR(75MHz,MeOD)δ 171.2,163.7(2 x C=O urea,amide),158.9,158.0(C=O Cbz,C イプソPhenol), 138.3, 130.8, 130.3, 129.5, 129.0, 128.8, 127.8, 121.8, 112.9 (C-Ar), 73.0, 70.4, 69.18, 68.21, 68.1, 67.5, 55.2, 55.1, 50.4, 47.1, 41.5, 40.5, 30.4, 30.2.C 29 H 37 N4O7 + [M+H] + Calculated for LC-HRMS (ESI): 553.2657; Found: 553.2665.

[0104] Unless otherwise indicated, the following examples were synthesized similarly to the synthesis described for Example 1, using the appropriate building blocks. [Table 2] TIFF2025530870000071.tif229169 TIFF2025530870000072.tif193169 TIFF2025530870000073.tif214169 TIFF2025530870000074.tif67169

[0105] Example 15 Benzyl (2-(2-(((7-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-7-azaspiro[3.5]nonan-2-yl)oxy)methyl)phenoxy)ethyl)carbamate [ka] Step g tert-Butyl 2-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)-7-azaspiro[3.5]nonane-7-carboxylate (BB13) [ka] In a similar manner to the procedure described in Example 1g), benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate (see Example 1) was reacted with tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate (CAS RN240401-28-9) to give the title compound (59.6 mg, 0.114 mmol, 76%). 1 H NMR(300MHz,CDCl3)δ 7.38-7.26(m,7H),6.95(td,J=7.5,1.0Hz,1H),6.85(d,J=8.2Hz,1H),5.57(s,1H,NHCbz),5.10(s,2H,OCH2),4.41(s,2H,OCH2), 4.16-3.95(m,3H),3.61(q,J=5.3Hz,2H),3.23(dt,J=20.9,5.7Hz,4H),2.12(t,J=10.0Hz,2H),1.80-1.57(m,3H),1.45(m,12H). 13 C NMR(75MHz,CDCl3)δ 156.7,155.1(C=O Cbz,C=O Boc,C Ar -O),136.6,130.1,129.4,128.7,128.4,128.3,126.9,121.2,112.2,79.4(C(CH3 )3Boc),69.1,67.7,67.0,65.7,41.0,40.9,40.8,39.6,39.4,36.7,31.0,28.6(3 x CH3Boc),27.1.C 30 H 40 N2O6Na[M+Na] + Calculated LC-HRMS (ESI): 547.2779; Found: 547.2799.

[0106] Step a1 Benzyl (2-(2-(((7-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-7-azaspiro[3.5]nonan-2-yl)oxy)methyl)phenoxy)ethyl)carbamate 1. Boc Deprotection

[0077] Analogously to the procedure described in Example 1a1, step (1), tert-butyl 2-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)-7-azaspiro[3.5]nonane-7-carboxylate (BB13) (52.5 mg, 0.10 mmol) was dissolved in DCM (0.7 mL) at 0°C, and TFA (0.11 mL, 1.40 mmol) was added. The reaction mixture was stirred at that temperature until Boc deprotection was complete (30 min, LC-MS monitoring). The reaction mixture was diluted with toluene (3.00 mL) and coevaporated twice with toluene (3.00 mL) under reduced pressure. The trifluoroacetate salt of the free amine was used in the next step without further purification. 2. Urea Coupling Triphosgene (CAS RN32315-10-9, 53.8 mg, 0.10 mmol) and NaHCO (33.6 mg, 0.08 mmol) were suspended in DCM (1.0 mL) at 0 °C. The deprotected amine derivative of benzyl (2-(2-(((7-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-7-azaspiro[3.5]nonan-2-yl)oxy)methyl)phenoxy)ethyl)carbamate obtained in step a1, 1) was dissolved in anhydrous DCM (1.5 mL) and added dropwise to the triphosgene mixture. The mixture was allowed to warm to room temperature and stirred overnight. The filtrate from this solution was then added dropwise to a solution of (4aR,8aS)-hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one (CAS RN2377107-31-6, 15.6 mg, 0.10 mmol) and DIPEA (70.0 μL, 0.40 mmol) in DCM (1.0 mL) at 0 °C. The mixture was warmed to room temperature and stirred for an additional 3 h. The mixture was concentrated under reduced pressure and purified by RP-HPLC (25-75% ACN in HO containing 0.1% TFA) to obtain the title compound (23.6 mg, 39.0 μmol, 39%). 1H NMR(300MHz,MeOD)δ 7.37-7.19(m,7H),6.91(td,J=8.3,1.4Hz,2H),5.08(s,2H,OCH2),4.42( s,2H,OCH2),4.24-4.00(m,5H),3.96(d,J=3.0Hz,1H),3.59(dd,J=12.7,5 .1Hz,1H),3.52(t,J=5.4Hz,2H),3.44-3.32(m,2H),3.17-2.89(m,6H),2. 21-2.09(m,2H),1.92-1.82(m,2H),1.77-1.64(m,2H),1.56-1.41(m,4H). 13 C NMR(75MHz,CDCl3)δ 171.2,165.5,158.9,158.1(C=O urea,C=O amide,C=O Cbz,C Ar -O),138.4,130.9,130.3,129.5,129.0,128.7,127.9,121.8,112.8,70.6,70.3 ,68.2,67.5,66.2,50.4,45.2,45.1,42.9,41.6,40.6,40.4,37.7,32.3,30.3.C 33 H 42 N4O7Na[M+Na] + Calculated for LC-HRMS (ESI) 629.2946; Found: 629.2961.

[0107] Unless otherwise indicated, the following examples were synthesized similarly to the synthesis described for Example 15, using the appropriate building blocks. [Table 3]

[0108] Example 17 Benzyl (R)-(2-(2-(((1-(3-(2-oxooxazolidin-4-yl)propanoyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate [ka] Building Block Synthesis: 3-[(4R)-2-Oxooxazolidin-4-yl]propanoic Acid (3 Steps) Step 1) Methyl (S)-4-((tert-butoxycarbonyl)amino)-5-hydroxypentanoate To a solution of (R)-2-((tert-butoxycarbonyl)amino)-5-methoxy-5-oxopentanoic acid (1 g, 3.83 mmol, CAS: 76379-01-6) in THF (15 mL) was added N-methylmorpholine (421 μL, 3.83 mmol) at −10 °C, followed by ethyl chloroformate (368 μL, 3.83 mmol), and the reaction mixture was stirred at this temperature for 10 min. NaBH (434 mg, 11.5 mmol) was added in one portion without causing a temperature increase. MeOH (35 mL) was added dropwise over 30 min at −1 °C to 17 °C. Stirring was continued for 1 h in an ice bath. 1 M aqueous KHSO sol (40 mL) was added dropwise to the reaction mixture, and then the organic solvent was evaporated under reduced pressure. The aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with 1M aqueous KHSO4 and saturated aqueous NaHCO3, dried over MgSO4, filtered, loaded onto silica gel, and evaporated under reduced pressure. The crude compound was purified by silica gel chromatography using an MPLC system eluting with a gradient of n-heptane:ethyl acetate (100:0 to 30:70) to give the title compound as a colorless oil (0.70 g, 66%). MS (ESI): m / z = 192.1 [M+H] + .

[0109] Step 2) Methyl (R)-3-(2-oxooxazolidin-4-yl)propanoate To a solution of methyl (S)-4-((tert-butoxycarbonyl)amino)-5-hydroxypentanoate (690 mg, 2.79 mmol, 1.0 equiv.) in THF (8.8 mL) was added thionyl chloride (611 μL, 8.37 mmol, 3.0 equiv.) dropwise, and the solution was stirred at room temperature for 3 h. Silica gel was added, and the reaction mixture was evaporated under reduced pressure. The compound was purified by silica gel chromatography using an MPLC system eluting with a gradient of n-heptane:ethyl acetate (100:0 to 0:100) to give the title compound as a colorless oil (404 mg, 79%). MS (ESI): m / z = 174.1 [M+H] + .

[0110] Step 3: (R)-3-(2-oxooxazolidin-4-yl)propanoic acid To a solution of (S)-methyl 3-(2-oxooxazolidin-4-yl)propanoate (400 mg, 2.31 mmol, 1.0 equiv.) in 1,4-dioxane (2 mL) and water (2 mL), lithium hydroxide monohydrate (107 mg, 2.54 mmol, 1.1 equiv.) was added, and the reaction mixture was stirred at room temperature for 2 hours. 1,4-Dioxane was evaporated, and aqueous HCl (2.54 mL, 2.54 mmol, 1.1 equiv.) was added dropwise to the solution. The aqueous layer was extracted five times with ethyl acetate. The combined organic layers were dried over MgSO4, filtered, and evaporated to give the title compound as a colorless solid (330 mg, 86%). MS (ESI): m / z = 160.1 [M+H] + . Step a2 (R)-(2-(2-(((1-(3-(2-oxooxazolidin-4-yl)propanoyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate (R)-3-(2-oxooxazolidin-4-yl)propanoic acid (12.7 mg, 0.08 mmol) was dissolved in 1:1 THF / ACN (1.60 mL), and then DIPEA (42.0 μL, 0.24 mmol) was added, followed by HATU (36.5 mg, 96.0 μmol). After stirring for 10 min, the deprotected amine derivative of tert-butyl 3-(((2-(2-((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carboxylate BB1 ​​(38.7 mg, 0.08 mmol, prepared as described in Example 1, Step 1, Boc deprotection) was added. The mixture was stirred at room temperature overnight, then the mixture was diluted directly with ACN / HO and purified by RP-HPLC (15-85% ACN in HO containing 0.1% TFA) to give the title compound (16.3 mg, 0.032 mmol, 40%). 27 H 34 N3O7[M+H] + Calculated for LC-HRMS (ESI): 512.2389; Found: 512.2391.

[0111] Example 18 Benzyl (2-(2-(((1-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate [ka] In a similar manner to the procedure described in Example 17, step a2, tert-butyl 3-(((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carboxylate BB1 ​​was reacted with 3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6-carboxylic acid (CAS RN: 134997-87-8) to give the title compound (22.3 mg, 0.0.41 mmol, 50%). 30 H 32 N3O7[M+H] +Calculated for LC-HRMS (ESI): 546.2235; Found: 546.2239.

[0112] Example 19 2-(6-(2-(2-((7-nitro-1l2,3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one [ka] Step b 2-(6-(2-(2-aminoethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one [ka] A mixture of 20% tBuOH in EtOAc (HPLC grade) was purged with N for at least 15 minutes and maintained under a N atmosphere in a sealed vial. In a pear-shaped flask, the starting material, benzyl (2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate (Example 13, 16.0 mg, 0.03 mmol), was dissolved in the previously prepared solvent mixture and purged with N for 10 minutes. Palladium on carbon (10 mol%) was then added to the mixture, which was then purged with N for an additional 15 minutes. The reaction was then carried out using a H-filled balloon to constantly bubble H through the solution until completion (LC-MS monitoring). The solvent mixture level was maintained at 1-2 mL by adding small amounts of the previously prepared N2-purged solvent mixture as needed. Palladium on carbon was removed from the mixture by filtering through a PTFE syringe filter (pore size: 0.45 μm) using ACN and water, washing the filter. The crude product was lyophilized, and the resulting title compound was used in the next step (step c) without further purification. LC-MS [M+H] + :400.9.

[0113] Step c 2-(6-(2-(2-((7-nitro-1l2,3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one 2-(6-(2-(2-aminoethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one (12.0 mg, 0.03 mmol) from step c) was dissolved in MeOH (0.75 mL) and trimethylamine (13.0 μL, 0.09 mmol) was added. Then, a solution of 4-fluoro-7-nitrobenz-2-oxa-1,3-diazole (CAS RN29270-56-2, 11.0 mg, 0.06 mmol) in MeOH (0.3 mL) was added dropwise. The reaction was stirred in the dark for 3.5 h. The solvent was then removed under reduced pressure at 20° C. and the reaction was purified by RP-HPLC with 15-85% ACN in H 2 O containing 0.1% TFA to give the title product (3.19 mg, 6.0 μmol, 19%). 1 H NMR (600 MHz, CDCN) δ 8.53(d,J=8.8Hz,1H),7.16(td,J=7.8,1.7Hz,1H),7.06(dd,J=7.4,1.7Hz,1H), 6.92(dd,J=8.3,1.1Hz,1H),6.87(td,J=7.4,1.1Hz,1H),6.51-6.39(m,2H),4.4 7(s,2H),4.27(t,J=5.0Hz,2H),4.05-3.83(m,6H),3.67(s,2H),3.60(s,2H),2. 55(d,J=7.5Hz,2H),2.26(p,J=7.8Hz,1H),2.03-1.96(m,2H),1.75-1.68(m,2H). 13 C NMR(151MHz,CD3CN)δ 162.19,157.44,156.29,144.81,137.33,130.22,129.00,127.24,120.80,11 1.37,74.18,65.81,62.84,62.43,61.60,54.21,38.31,36.10,34.53,29.56.C 27 H 30 N7O7[M+H] + Calculated for LC-HRMS (ESI): 564.2201; Found: 564.2192

[0114] Unless otherwise indicated, the following examples were synthesized similarly to the synthesis described for Example 19, using the appropriate building blocks. [Table 4] TIFF2025530870000083.tif225169

[0115] Example 26 2-(7-(dimethylamino)-3-(dimethyliminio)-5,5-dimethyl-3,5-dihydrodibenzo[b,e]silylin-10-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate [ka] Step b 2-(6-(2-((5-aminopentyl)oxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one [ka] In a similar manner to the procedure described in Example 19b), benzyl (5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)pentyl)carbamate (Example 14) was treated with hydrogen and palladium / carbon to give the title compound, which was used in the next step (step c) without further purification. LC-MS [M+H] + :442.9.

[0116] Step c 2-(7-(dimethylamino)-3-(dimethyliminio)-5,5-dimethyl-3,5-dihydrodibenzo[b,e]silyn-10-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate 2-(6-(2-((5-aminopentyl)oxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one (8.96 mg, 0.02 mmol) was dissolved in ACN containing 10% DMF (0.90 mL) at 0 °C, followed by the addition of 2-(7-(dimethylamino)-3-(dimethyliminio)-5,5-dimethyl-3,5-dihydrodibenzo[b,e]silyn-10-yl)-4-(((2,5-dioxopyrrolidin-1-yl)oxy)carbonyl)benzoate (CAS RN1426090-09-6, 5.00 mg, 9.0 μmol), followed by DIPEA (9.0 μL, 7.0 μmol). The reaction was stirred at room temperature in the dark for approximately 4 hours (LC-MS monitoring of the reaction). It was then diluted with HO / ACN and lyophilized. The crude product was purified by RP-HPLC with 25-75% HO / ACN containing 0.1% TFA over 30 minutes to give the title product (6.0 mg, 7.0 μmol, 76%). 1H NMR(600MHz,MeOD)δ 8.33(d,J=8.2Hz,1H),8.14(dd,J=8.2,1.8Hz,1H),7.74(d,J=1.7Hz,1H),7.36(d,J=2.9Hz,2H) ,7.13(td,J=7.8,1.7Hz,1H),7.04(dd,J=7.4,1.7Hz,1H),7.02-6.99(m,2H),6.88(dd,J=8.2,1 .1Hz,1H),6.83(td,J=7.3,1.0Hz,1H),6.77(dd,J=9.6,2.9Hz,2H),4.53(s,2H,OCH2),4.07(s, 4H,OCH2),4.00(t,J=6.1Hz,2H),3.93(s,2H),3.84(s,2H),3.47(t,J=7.1Hz,2H),3.32(s,12H,4 x N-CH3), 2.64 (d, J = 7.4 Hz, 2H), 2.44 (p, J = 7.7 Hz, 1H, CH spirocycle), 2.22-2.16 (m, 2H, CH-CH2-C spirocycle), 1.88 (ddd, J = 10.5, 7.9, 2.8 Hz, 4H, CH-CH2-C spirocycle, -CH2-alkyl), 1.74 (p, J = 7.2 Hz, 2H, -CH2-alkyl), 1.67-1.59 (m, 2H, -CH2-alkyl), 0.68 (s, 3H, Si-CH3), 0.62 (s, 3H, Si-CH3). 13 C NMR (151 MHz, MeOD) δ 166.6, 162.3, 159.0, 156.8 (C=O urea, C=O carbamate, COO - ,C Ar -O), 129.7, 128.5, 127.4, 126.9, 120.1, 119.8, 113.8, 110.9, 74.2 (OCH2), 67.2 (OCH2), 63.0, 62.4, 61.8, 54.4 (-C-carbamate spirocycle), 39.8, 39.6, 38.2, 36.0, 34.7 (-C-spirocycle), 29.7 (CH spirocycle), 28.9, 28.8 (-CH2-alkyl), 23.5 (-CH2-alkyl), -2.20 (Si-CH3), -3.09 (Si-CH3). 51 H 60 N6O7Si[M+H] +Calculated for LC-HRMS (ESI): 897.4366; Found: 897.4357.

[0117] Unless otherwise indicated, the following examples were synthesized similarly to the synthesis described for Example 26, using the appropriate building blocks.

[0118] Example 32 is a mixture of positional isomers of 5- and 6-carboxytetramethylrhodamine derivatives obtained in one reaction. A fraction of the 5-carboxytetramethylrhodamine derivative (Example 33) was separated from the mixture by HPLC purification. Each 6-carboxytetramethylrhodamine derivative was not obtained in a pure form. The same considerations apply to Examples 34 and 35, and 36 and 37, respectively. Example 38 was obtained as a mixture of positional isomers of 5- and 6-carboxytetramethylrhodamine derivatives. [Table 5] TIFF2025530870000087.tif223169 TIFF2025530870000088.tif103169 TIFF2025530870000089.tif165169 TIFF2025530870000090.tif124169 TIFF2025530870000091.tif175169 TIFF2025530870000092.tif124169 TIFF2025530870000093.tif160169 TIFF2025530870000094.tif124169 TIFF2025530870000095.tif165169

[0119] Example 39 Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate [ka] Step d tert-Butyl 3-((3-(ethoxycarbonyl)phenoxy)methyl)azetidine-1-carboxylate (BB15) [ka] In a similar manner to the procedure described in Example 1d), ethyl 3-hydroxybenzoate (CAS RN7781-98-8) was reacted with tert-butyl 3-(bromomethyl)azetidine-1-carboxylate (CAS RN253176-93-1) to give the title compound (18.0 mg, 0.05 mmol, 18%). LC-MS (ESI) [M+Na] + .[M+Na] + :338.1

[0120] Step a1 Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate Analogously to the procedure described in Example 1, a1), (4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazin-6-ium(2S,3S)-3-carboxy-2,3-bis((4-methylbenzoyl)oxy)propanoate salt (CAS RN2624363-49-9, 66.7 mg, 0.09 mmol) was reacted with the deprotected amine derivative of tert-butyl 3-((3-(ethoxycarbonyl)phenoxy)methyl)azetidine-1-carboxylate BB15 (33.5 mg, 0.10 mmol, prepared as described for Example 1, Step 1, Boc deprotection) to give the title compound (16.3 mg, 0.032 mmol, 40%). LC-MS (ESI) [M+H] + :418.2.

[0121] Unless otherwise indicated, the following examples were synthesized similarly to the synthesis described for Example 39, using the appropriate building blocks. [Table 6]

[0122] Example 42 tert-Butyl (2-(3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzamido)ethyl)carbamate [ka] Process K: 1. Ester Hydrolysis: Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoate (Example 40, 12.5 mg, 0.03 mmol) was dissolved in THF / MeOH / HO:1:1 (0.2 mL) and lithium hydroxide (LiOH, 2, 15 mg, 0.09 mmol) was added. The reaction was stirred at room temperature until complete (approximately 2 hours). The reaction was quenched by diluting with ethyl acetate (1.0 mL), adding 1 M aqueous HCl (0.5 mL), and washing with brine (0.5 mL). The organic layer was dried over MgSO, filtered, and the solvent removed under reduced pressure. The carboxylic acid 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoic acid was used in the next step without further purification. 2. Amide Coupling. Crude 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoic acid, obtained in step 1 from ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoate (Example 40), was dissolved in DMF (0.43 mL) and DIPEA (16.0 μL, 0.09 mmol), followed by 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, CAS RN 0221-10-1). To the resulting solution was added 148893-10-1 (13.7 mg, 0.04 mmol). The mixture was stirred for 10 minutes, and then tert-butyl (2-aminoethyl)carbamate (CAS RN 57260-73-8, 5.28 mg, 0.03 mmol). The mixture was stirred at room temperature for 18 hours, then diluted with ethyl acetate (1.0 mL), washed with brine (0.5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by RP-HPLC H2O / ACN containing 0.1% TFA 25-75% to give the title compound (5.0 mg, 0.01 mmol, 32%). 25 H 36 HRMS calculated for N2O7[M+H]+ (ESI): 518.2609; Found: 518.2601

[0123] Example 43 tert-Butyl (2-(2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzamido)ethyl)carbamate [ka] Analogously to the procedure described in Example 42, k) ethyl 2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate (Example 41) was subjected to ester hydrolysis (step k,1) and then reacted with tert-butyl (2-aminoethyl)carbamate (CAS RN57260-73-8, 5.28 mg, 0.03 mmol) to give the title compound (5.0 mg, 9.0 μmol, 55%). LC-MS (ESI) [M+H] + :532.2.

[0124] Example 44 (4aR,8aS)-6-(3-((benzyloxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one [ka] Step 1t-butyl 3-((benzyloxy)methyl)azetidine-1-carboxylate [ka] tert-Butyl 3-(hydroxymethyl)azetidine-1-carboxylate (CAS RN 142253-56-3, 103 mg, 0.55 mmol) and a 60% dispersion of sodium hydride in mineral oil (CAS RN 7646-69-7, 55.0 mg, 1.38 mmol) were suspended in anhydrous DMF (2.0 mL) at 0 °C for 15 minutes or until no more gas evolution was observed. Benzyl bromide (CAS RN 100-39-0, 0.16 mL, 1.38 mmol) was then added. The reaction was allowed to warm to room temperature and stirred for 16 hours. The mixture was diluted with dichloromethane (10.0 mL), neutralized with acetic acid, saturated NaHCO (10 mL) was added, and washed with water (10 mL). The organic layer was dried over MgSO, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel flash chromatography using 0-80% ethyl acetate in cyclohexane as eluent to give the title compound (120 mg, 0.43 mmol, 77%). 1 H NMR(300MHz,CDCl3)δ 7.40-7.26(m,5H),3.99(t,J=8.5Hz,2H),3.66(dd,J=8.7,5.3Hz,2H),3.58(d,J=6.8Hz,2H),2.88-2.67(m,1H),1.43(s,9H).LC-MS[M+Na] + :300.1.

[0125] Step a1 (4aR,8aS)-6-(3-((benzyloxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one In a similar manner to the procedure described in Example 1, a1), tert-butyl 3-((benzyloxy)methyl)azetidine-1-carboxylate was reacted with (4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazin-6-ium(2S,3S)-3-carboxy-2,3-bis((4-methylbenzoyl)oxy)propanoate salt (CAS RN2624363-49-9) to give the title compound. LC-MS (ESI) [M+H] + :360.1.

[0126] Example 45 (4aR,8aS)-6-(6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one [ka] Step 1 tert-Butyl 6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carboxylate [ka] Analogously to the procedure described in Example 44l), tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate (CAS RN 1147557-97-8) was reacted with benzyl bromide (CAS RN 100-39-0) to give the title compound. 1 H NMR(300MHz,CDCl3)δ 7.41-7.27(m,5H),4.39(s,2H),4.02-3.75(m,5H),2.46(ddd,J=9.8,6.8,3.0Hz,2H),2.14(ddd,J=10.1,7.3,3.0Hz,2H),1.42(s,9H).

[0127] Step a1 (4aR,8aS)-6-(6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one In a similar manner to the procedure described in Example 1, a1), tert-butyl 6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carboxylate was reacted with (4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazin-6-ium(2S,3S)-3-carboxy-2,3-bis((4-methylbenzoyl)oxy)propanoate salt (CAS RN 2624363-49-9) to give the title compound. LC-MS (ESI) [M+H] + :386.2.

[0128] Example 46 (4aR,8aS)-6-(3-(phenoxymethyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one [ka] In a similar manner to the procedure described in Example 1 a1), tert-butyl 3-(phenoxymethyl)azetidine-1-carboxylate (obtained in a similar manner to Example 1d from phenol, CAS RN108-95-2, and tert-butyl 3-(bromomethyl)azetidine-1-carboxylate, CAS RN 253176-93-1) was reacted with (4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazin-6-ium(2S,3S)-3-carboxy-2,3-bis((4-methylbenzoyl)oxy)propanoate salt (CAS RN2624363-49-9) to give the title compound. LC-MS (ESI) [M+H] + :346.1

[0129] Composition of Components BB2 tert-Butyl 3-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)azetidine-1-carboxylate [ka] Analogously to the procedure described in Example 1g), tert-butyl 3-hydroxy-azetidine-1-carboxylate (CAS RN 141699-55-0, 125 mg, 0.72 mmol) was reacted with benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate (see Example 1, 131 mg, 0.36 mmol) to give the title compound (101 mg, 0.22 mmol, 61%). 1H NMR(300MHz,CDCl3)δ 7.41-7.27(m,7H),6.97(t,J=7.4Hz,1H),6.86(d,J=8.1Hz,1H),5.43(s,1H) ,NH-Cbz),5.11(s,2H,OCH2-Ar),4.46(s,2H,OCH2Cbz),4.31(dq,J=10.8,5 .1Hz,1H,CH azetidine),4.10(t,J=5.0Hz,2H),4.07-3.97(m,2H),3.92-3.81(m ,2H),3.62(q,J=5.4Hz,2H,OCH2CH2-NHCbz),1.42(d,J=1.0Hz,9H,CH3Boc). 13 C NMR(75MHz,CDCl3)δ 156.6,156.5(C=O Cbz,C=O urea),136.6,130.1,129.7,128.7,128.4,128.3,121.3,112.0(C-Ar),79.7(CH Boc),67.6,67.0,66.5(3 x OCH2),56.65,40.8(CH2-NHCbz),28.5(CH3Boc).C 25 H 33 N2O6[M+H] + Calculated for LC-HRMS (ESI): 457.2333; Found: 457.2320.

[0130] BB3 tert-Butyl 6-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)-2-azaspiro[3.3]heptane-2-carboxylate [ka] In a similar manner to the procedure described in Example 1g), benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate (see Example 1) was reacted with tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate (CAS RN1147557-97-8) to give the title compound (48.5 mg, 0.098 mmol, 65%). 1H NMR (300 MHz, CDCl3) δ 7.39-7.21 (m, 7H), 6.95 (td, J = 7.5, 1.0 Hz, 1H), 6.84 (d, J = 8.5 Hz, 1H), 5.54 (s, 1H, NHCbz), 5.10 (s, 2H, OCH2), 4.40 (s, 2H, OCH2), 4.10 (t, J = 5.0 Hz, 2H), 3.93-3.72 (m, 5H, CH spirocycle, 2 x NCH2 spirocycle), 3.61 (q, J = 5.3 Hz, 2H, CH2NHCbz), 2.41-2.28 (m, 2H, C-CH2-CH spirocycle), 2.14-1.95 (m, 2H, C-CH2-CH spirocycle), 1.43 (s, 9H, 3 x CH3Boc). 13 C NMR(75MHz,CDCl3)δ 156.7,156.6,156.2(C=O Cbz,C=O Boc,C Ar -O), 136.5, 130.1, 129.5, 128.7, 128.5, 128.4, 126.6, 121.2, 112.1, 79.4 (C(CH3)3Boc), 68.1, 67.6, 67.0, 65.8, 61.8, 60.5, 41.2, 40.7 (CH2NHCz, 2 x C-CH2-CH spirocycle), 30.1 (-C-spirocycle), 28.5 (3 x CH3Boc).C 28 H 36 N2O6Na[M+Na] + Calculated for: 519.2466; Measured: 519.2450.

[0131] BB4 tert-Butyl 7-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)-2-azaspiro[3.5]nonane-2-carboxylate [ka] In a similar manner to the procedure described in Example 1g), benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate (see Example 1) was reacted with tert-butyl 7-hydroxy-2-azaspiro[3.5]nonane-2-carboxylate (CAS RN 1363383-18-9) to give the title compound (18.0 mg, 0.05 mmol, 26%). 1 H NMR(300MHz,CDCl3)δ 7.35(s,7H),7.02-6.77(m,2H),5.41(s,1H,NHCbz),5.10(s,2H,OCH2),4.53(s,2H,OCH2),4.08 (t,J=5.0Hz,2H),4.04-3.93(m,1H),3.69-3.41(m,7H),1.94-1.72(m,4H),1.54-1.32(m,12H). 13 C NMR(75MHz,CDCl3)δ 156.6,156.5,156.2(C=O Cbz,C=O Boc,C Ar -O),136.4,129.5,128.9,128.6,128.5,128.2,128.1,127.9,127.4,121.1,111.7,7 9.2(C(CH3)3Boc),67.3,66.9,65.2,59.0,40.7(CH2NHCbz),34.5,33.0,28.5,28.2(3 x CH3Boc).C 30 H 40 N2O6Na[M+Na] + Calculated for LC-HRMS (ESI): 547.2779; Found: 547.2774.

[0132] BB5 tert-Butyl 3-(((3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carboxylate [ka] Step d Ethyl 3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzoate [ka] In a similar manner to the procedure described in Example 1d), ethyl 3-hydroxybenzoate (CAS RN7781988) was reacted with 2-(((benzyloxy)carbonyl)amino)ethyl 4-methylbenzenesulfonate (CAS RN 93407-96-6) to give the title compound (1.05 g, 3.05 mmol, 61%). 1 H NMR(300MHz,CDCl3)δ 7.66(dt,J=7.7,1.3Hz,1H),7.54(dd,J=2.7,1.5Hz,1H),7.40-7.27(m,6H),7.12-7.02(m,1H),5.22(s,1H,NH-Cbz),5.12(s,2H,OC H2CH3),4.37(q,J=7.1Hz,2H,OCH2),4.09(t,J=5.1Hz,2H,OCH2),3.63(q,J=5.4Hz,2H,CH2NHCbz),1.39(t,J=7.1Hz,3H,OCH2CH3). 13 C NMR(75MHz,CDCl3)δ 166.5,158.5,156.5(C=O CO2Et,C=O Cbz,C Ar -O),136.5,132.1,129.6,128.7,128.4,128.3,122.6,119.7,114.9,67.3(OCH2),67.1(OCH2),61.3(OCH2),40.7(CH2NHCbz),14.5(CH3).C 19 H 22 NO5[M+H] + Calculated for LC-HRMS (ESI) 366.1492; Found: 366.1508.

[0133] Step e Benzyl (2-(3-(hydroxymethyl)phenoxy)ethyl)carbamate [ka] Ethyl 3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzoate was reacted in a similar manner to the procedure described in Example 1e) to give the title compound (115 mg, 0.38 mmol, 66%). 1H NMR(300MHz,CDCl3)δ 7.43-7.16(m,6H),7.00-6.85(m,2H),6.87-6.70(m,1H),5.11(s,2H,O-CH2-),4.66(s,2H) ,O-CH2-),4.04(t,J=5.1Hz,2H,-OCH2CH2NHCbz),3.60(d,J=5.3Hz,2H,-OCH2CH2NHCbz). 13 C NMR(75MHz,CDCl3)δ 158.8,156.6(C=O Cbz,C Ar -O),142.8,136.5,129.8,128.7,128.32,128.28,119.7,113.8,112.9(C- Ar),67.01(O-CH2-),66.96(O-CH2-),65.2(O-CH2-),40.7(-CH2-NHCbz).C 17 H 20 NO4[M+H] + Calculated for LC-HRMS (ESI): 302.1387; Found: 302.1373. Step f Benzyl (2-(3-(bromomethyl)phenoxy)ethyl)carbamate [ka] Benzyl (2-(3-(hydroxymethyl)phenoxy)ethyl)carbamate was reacted in a similar manner to the procedure described in Example 1f) to give the title compound (503 mg, 1.38 mmol, 63%). 1 H NMR(300MHz,CDCl3)δ 7.42-7.22(m,6H),7.03-6.97(m,1H),6.95-6.90(m,1H),6.88-6.79(m,1H),5.24(s,1H,NHCbz),5. 13(s,2H,OCH2),4.46(s,2H,CH2Br),4.06(t,J=5.1Hz,2H,OCH2),3.63(q,J=5.4Hz,2H,CH2NHCbz). 13 C NMR(75MHz,CDCl3)δ 158.8,156.5(C=O Cbz,C Ar-O),139.4,139.2,136.5,130.1,128.7,128.33,128.29,121.9,121.4,115.2,114.7,114.7,67.0(2 x OCH2),40.7(CH2NHCbz),33.4(CH2Br).C 17 H 19 BrNO3[M+H] + Calculated for LC-HRMS (ESI): 366.0524; Found: 366.0551.

[0134] Step g tert-Butyl 3-(((3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carboxylate In a similar manner to the procedure described in Example 1g), benzyl (2-(3-(bromomethyl)phenoxy)ethyl)carbamate was reacted with tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (CAS RN 142253-56-3) to give the title compound (85.0 mg, 0.18 mmol, 55%). 1 H NMR(300MHz,CDCl3)δ 7.40-7.20(m,6H),7.01-6.74(m,3H),5.31-5.06(m,3H,NHCbz,OCH2),4.70-4.42(m,3H), 4.07-3.91(m,4H),3.74-3.51(m,5H),2.88-2.69(m,1H,CH azetidine),1.43(d,J=1.2Hz,9H,3 x CH3Boc).C 26 H 34 N2O6Na[M+Na] + Calculated for LC-HRMS (ESI): 493.2309; Found: 493.2312

[0135] BB6 tert-Butyl 3-((3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)azetidine-1-carboxylate [ka] In a similar manner to the procedure described for BB7, benzyl (2-(3-(bromomethyl)phenoxy)ethyl)carbamate was reacted with tert-butyl 3-hydroxyazetidine-1-carboxylate (CAS RN 141699-55-0) to give the title compound (55.1 mg, 0.12 mmol, 55%). 25 H 33 N2O6[M+H] + Calculated for LC-HRMS (ESI): 457.2333; Found: 457.2328.

[0136] BB7 tert-Butyl 6-((3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)-2-azaspiro[3.3]heptane-2-carboxylate [ka] In a similar manner to the procedure described for BB7, benzyl (2-(3-(bromomethyl)phenoxy)ethyl)carbamate was reacted with tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate (CAS RN 1147557-97-8) to give the title compound (21.0 mg, 0.042 mmol, 28%). 1 H NMR (300 MHz, CDCl3) δ 7.36 (d, J = 4.9 Hz, 5H), 7.22 (d, J = 7.8 Hz, 1H), 6.93-6.74 (m, 3H), 5.11 (s, 2H, OCH2), 4.35 (s, 2H, OCH2), 4.04 (t, J = 5.1 Hz, 2H, OCH2), 4.01-3.78 (m, 5H, N-CH2-C spirocycle, CH spirocycle), 3.61 (q, J = 5.4 Hz, 2H, CH2NHCbz), 2.47 (m, 2H, C-CH2-CH spirocycle), 2.14 (m, 2H, C-CH2-CH spirocycle), 1.42 (s, 9H, 3 x CH3Boc). 13 C NMR(75MHz,CDCl3)δ 158.7,156.5,156.3(C=O Cbz,C=O Boc,C Ar-O), 139.9, 136.5, 129.7, 128.7, 128.34, 128.30, 120.6, 113.8, 79.5 (C(CH3)3Boc), 70.1, 68.1, 67.0, 61.9, 60.6, 41.2, 40.7, 30.3 (3 x CH3Boc), 28.5 (-C-spirocycle).C 28 H 37 N2O6[M+H] + Calculated for LC-HRMS (ESI): 497.2646; Found: 497.2636.

[0137] BB8 tert-Butyl 3-(((2-(3-(((benzyloxy)carbonyl)amino)propoxy)benzyl)oxy)methyl)azetidine-1-carboxylate [ka] Step d Ethyl 2-(3-(((benzyloxy)carbonyl)amino)propoxy)benzoate [ka] In a similar manner to the procedure described in Example 1d), ethyl salicylate (CAS RN 118-61-6) was reacted with 3-(((benzyloxy)carbonyl)amino)propyl 4-methylbenzenesulfonate (CAS RN 68076-37-9) to give the title compound (3.98 g, 11.2 mmol, 82%). 1 H NMR(300MHz,CDCl3)δ 7.85(dd,J=7.8,1.8Hz,1H),7.56-7.20(m,6H),7.06-6.87(m,2H),6.54(s,1H,NHCbz),5.11(s,2H,OCH2),4.24(q,J=7.1Hz,2H,O CH2CH3),4.13(t,J=5.5Hz,2H,OCH2),3.57-3.40(m,2H,CH2NHCbz),2.06(p,J=5.6Hz,2H,-CH2-),1.29(t,J=7.1Hz,3H,OCH2CH3). 13C NMR(75MHz,CDCl3)δ 166.0,158.7,157.0(C=O CO2Et,C=O Cbz,C Ar -O),137.1,133.9,132.0,128.4,128.0,127.9,120.4,119.7,112.7,68.2(O CH2),66.5(OCH2),61.0(OCH2),39.9(CH2NHCbz),29.1(-CH2-),14.4(CH3).C 20 H 23 LC-HRMS (ESI) calculated for NO5Na[M+Na]+: 380.1468; Found: 380.1474.

[0138] Step e Benzyl (3-(2-(hydroxymethyl)phenoxy)propyl)carbamate [ka] Ethyl 2-(3-(((benzyloxy)carbonyl)amino)propoxy)benzoate was reacted in a similar manner to the procedure described in Example 1e) to give the title compound (2.35 g, 7.45 mmol, 98%). 1 H NMR(300MHz,CDCl3)δ 7.39-7.19(m,7H),6.94(td,J=7.4,1.1Hz,1H),6.83(d,J=8.1Hz,1H),5.08(s,2H,-OCH2-),4.66(s,2H,-OCH2-),4.03(td,J =5.9,1.7Hz,2H,-OCH2CH2CH2NHCbz),3.40(t,J=6.3Hz,2H,-OCH2CH2CH2NHCbz),1.98(p,J=5.9Hz,2H,-OCH2CH2CH2NHCbz). 13 C NMR(75MHz,CDCl3)δ 156.7,156.6(C=O Cbz,C Ar -O), 136.6, 129.3, 129.0, 128.5, 128.09, 128.04, 120.7, 111.0 (C-Ar), 66.7 (OCH2), 65.7 (OCH2), 61.5 (OCH2), 38.6 (CH2-NHCbz), 29.4 (-CH2-linker).C 18 H 21NO4Na[M+Na] + Calculated for LC-HRMS (ESI): 338.1363; Found: 338.1353.

[0139] Step f Benzyl (3-(2-(bromomethyl)phenoxy)propyl)carbamate [ka] Benzyl (3-(2-(hydroxymethyl)phenoxy)propyl)carbamate was reacted in a similar manner to the procedure described in Example 1f) to give the title compound (2.00 g, 5.29 mmol, 80%). 1 H NMR(300MHz,CDCl3)δ 7.45-7.18(m,7H),6.98-6.79(m,2H),5.11(s,2H,OCH2),4.54(s,2H,CH2-Br),4.10(t,J= 5.8Hz,2H,,OCH2 linker),3.49(q,J=6.4Hz,2H,CH2-NHCbz),2.18-1.96(m,2H,-CH2-linker). 13 C NMR(75MHz,CDCl3)δ 156.8,156.7(C=O Cbz,C Ar -O), 136.7, 131.0, 130.8, 130.5, 130.4, 128.7, 128.2, 126.2, 125.9, 121.0, 111.77, 111.71 (C-Ar), 66.8 (-OCH2-), 65.85 (-OCH2-), 42.13 (-CH2-NHCbz), 38.6 (CH2-Br), 29.53 (-CH2- linker).C 18 H 21 BrNO3[M+H] + Calculated for LC-HRMS (ESI): 378.0699; Found: 378.0700

[0140] Step g tert-Butyl 3-(((2-(3-(((benzyloxy)carbonyl)amino)propoxy)benzyl)oxy)methyl)azetidine-1-carboxylate In a similar manner to the procedure described in Example 1g), benzyl (3-(2-(bromomethyl)phenoxy)propyl)carbamate was reacted with tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (CAS RN 142253-56-3) to give the title compound (401 mg, 0.83 mmol, 59%). 1 H NMR(300MHz,CDCl3)δ 7.41-7.28(m,7H),6.95(td,J=7.5,1.1Hz,1H),6.85(d,J=8.2Hz,1H),5.46 (s,1H,NHCbz),5.10(s,2H,OCH2),4.54(s,2H,OCH2),4.05(t,J=5.8Hz,2H) ,3.92(t,J=8.4Hz,2H),3.67-3.51(m,4H),3.43(q,J=6.1Hz,2H,CH2NHCbz) ,2.81-2.63(m,1H,CH azetidine),2.02(p,J=6.0Hz,2H,CH2 linker),1.42(s,9H,3 x CH3Boc). 13 C NMR(75MHz,CDCl3)δ 156.7,156.6,156.5(C=O Cbz,C=O Boc,C Ar -O), 136.8, 129.6, 129.2, 128.6, 128.2, 126.4, 120.8, 111.4, 79.4 (C(CH3)3Boc), 72.4 (OCH2), 68.7 (OCH2), 66.7 (OCH2), 66.4 (OCH2), 52.0 (2 x N-CH2-azetidine), 39.1 (CH2NHCbz), 29.6 (CH azetidine), 28.5 (3 x CH3Boc), 27.1 (-CH2-linker). 27 H 37 N2O6[M+H] + Calculated for LC-HRMS (ESI): 485.2646; Found: 485.2646.

[0141] BB9 tert-Butyl 3-(((2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzyl)oxy)methyl)azetidine-1-carboxylate [ka] Step d Ethyl 2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzoate [ka] In a similar manner to the procedure described in Example 1g), ethyl salicylate (CAS RN 118-61-6) was reacted with 5-(((benzyloxy)carbonyl)amino)pentyl 4-methylbenzenesulfonate (CAS RN 93066-51-4) to give the title compound (1.0 g, 2.59 mmol, 89%). 1 H NMR(300MHz,CDCl3)δ 7.77(dd,J=7.7,1.8Hz,1H),7.52-7.28(m,6H),7.04-6.84(m,2H),5.09(s,2H,O-CH2),4.31(q,J=7.1Hz,2H,O-CH2- CH3),4.02(t,J=6.2Hz,2H,O-CH2),3.23(m,2H,-CH2-NHCbz),1.84(p,J=6.5Hz,2H,-CH2-alkane),1.64-1.53(m,4H,2 x-CH2-alkane),1.33(t,J=7.1Hz,3H,O-CH2-CH3). 13 C NMR(75MHz,CDCl3)δ 166.6,158.6,156.6(C=O CO2Et,C=O Cbz,C Ar -O), 136.8, 133.4, 131.66, 128.6, 128.22, 128.19, 120.8, 120.3, 113.2 (C-Ar), 68.5, 66.7, 60.9 (3 x O-CH2), 41.0 (-CH2-NHCbz), 31.1, 28.8, 23.2 (3 x CH2 alkanes), 14.44 (O-CH2CH3).C 22 H 27 NO5Na[M+Na] + Calculated for LC-HRMS (ESI): 408.1781; Found: 408.1798. Step e Benzyl (5-(2-(hydroxymethyl)phenoxy)pentyl)carbamate [ka] Ethyl 2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzoate was reacted in a similar manner to the procedure described in Example 1e) to give the title compound (737 mg, 2.15 mmol, 86%). 1 H NMR(300MHz,CDCl3)δ 7.41-7.19(m,7H),6.93(td,J=7.4,1.0Hz,1H),5.10(s,2H,OCH2),4.68(s,2H,OCH2),4.01(t,J=6.3Hz,2H,OCH2),3.23(t,J=6.6Hz,2H CH2NHCbz),1.84(p,J=6.5Hz,2H,-CH2-alkane),1.68-1.46(m,4H,2 x-CH2-alkane). 13 C NMR(75MHz,CDCl3)δ 157.0,156.6(C=O Cbz,C Ar -O), 136.7, 129.3, 129.1, 128.9, 128.7, 128.3, 128.3, 120.8, 111.2, 67.7(OCH2), 66.8(OCH2), 62.3(CH2OH), 41.0(CH2NHCbz), 29.9(-CH2-alkane), 27.1(-CH2-alkane), 23.5(-CH2-alkane).C 20 H 25 NO4Na[M+Na] + Calculated for LC-HRMS (ESI): 366.1676; Found: 366.1691.

[0142] Step 1: ftrt-butyl 3-(((2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzyl)oxy)methyl)azetidine-1-carboxylate In a similar manner to the procedure described in Example 1f), benzyl (5-(2-(hydroxymethyl)phenoxy)pentyl)carbamate was reacted with tert-butyl 3-(hydroxymethyl)azetidine-1-carboxylate (CAS RN 142253-56-3) to give the title compound (67.6 mg, 0.132 mmol, 53%). 1H NMR(600MHz,CDCl3)δ 7.38-7.28(m,5H),7.27-7.19(m,2H),6.94(t,J=7.4Hz,1H),6.84(d,J=8.3Hz,1H) ,5.10(s,2H,OCH2),4.97(s,1H,NHCbz),4.56(s,2H,OCH2),4.00-3.93(m,4H),3.70 -3.64 (m, 2H), 3.64-3.59 (m, 2H), 3.23 (q, J = 6.7 Hz, 2H, CH2NHCbz), 2.82-2.73 (m, 1H, CH2azetidine), 1.85-1.77 (m, 2H, CH2 linker), 1.63-1.48 (m, 4H, CH2 linker), 1.43 (s, 9H, 3 x CH3Boc). 13 C NMR(151MHz,CDCl3)δ 156.63,156.58,156.55(C=O Boc,C=O Cbz,C Ar -O), 136.8, 129.1, 128.9, 128.6, 128.21, 128.17, 126.7, 120.5, 111.2, 79.4 (C(CH3)3Boc), 72.6, 68.2, 67.8, 66.7 (4 x OCH2), 52.0 (2 x NCH2 Azetidine), 41.1 (CH2NHCbz), 29.8, 29.0, 28.8 (CH Azetidine, 2 x CH2 Linker), 28.5 (3 x CH3Boc), 23.5 (CH2 Linker). LC-HRMS (ESI) calculated for C29H40N2O6Na [M+Na]+: 535.2779; found: 537.2784

[0143] BB10 tert-Butyl 6-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylate [ka] Step i Building Block Synthesis: tert-Butyl 6-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylate [ka] In an oven-dried flask, tetramethylpiperidine (CAS RN768-66-1, 1.25 mL, 7.40 mmol) was dissolved in anhydrous THF (37.0 mL) and cooled to -78 °C under a N atmosphere. A 2.5 M solution of nBuLi in THF (3.00 mL, 7.40 mmol) was added dropwise, and the reaction was stirred at the same temperature for 30 min. Next, a 0.84 M solution of bis((pinacolato)boryl)methane (CAS RN 78782-17-9) in THF (8.83 mL, 7.40 mmol) was added dropwise. The reaction was stirred for 5 min, and then a 0.20 M solution of the ketone in anhydrous THF (18.5 mL, 3.70 mmol) was added dropwise over 5 min. The reaction mixture was allowed to warm slowly to room temperature overnight. Upon completion, the reaction was opened to air and filtered through a silica plug eluting with EtO. The mixture was concentrated under reduced pressure and purified by silica gel chromatography using 0-20% EtOAc in cyclohexane with ELSD detection to give the title compound in quantitative yield. 18 H 31 BNO4[M+H] + Calculated for LC-HRMS (ESI): 336.2340; Found: 336.2342. Step d Benzyl (2-(2-bromophenoxy)ethyl)carbamate [ka] In a similar manner to the procedure described in Example 1d), 2-bromophenol (CAS 95-56-7) was reacted with 2-(((benzyloxy)carbonyl)amino)ethyl 4-methylbenzenesulfonate (CAS RN93407-96-6) to give the title compound (985 mg, 2.81 mmol, 94%). 1H NMR(300MHz,CDCl3)δ 7.47(dd,J=7.9,1.6Hz,1H),7.33-7.13(m,6H),6.87-6.73(m,2H),5.31(s,1H,NHCb z),5.06(s,2H,OCH2),4.03(t,J=5.0Hz,2H,OCH2),3.59(q,J=5.4Hz,2H,CH2NHCbz). 13 C NMR(75MHz,CDCl3)δ 156.6,154.9(C=O Cbz,C Ar -O), 136.5, 133.5, 128.7, 128.7, 128.29, 128.26, 122.6, 113.7, 112.5, 68.5 (OCH2), 67.0 (OCH2), 40.6 (CH2NHCbz). LC-HRMS (ESI) calculated for C16H16BrNO3Na[M+Na]+: 372.0206; found: 372.0209.

[0144] Step 1: tert-Butyl 6-((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylate In a sealed vial under a nitrogen atmosphere, tert-butyl 6-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-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 mixture of dioxane / water (2.30 mL) that had been pre-purged by bubbling N2 through it with sonication. The mixture was purged with N2 bubbling for 10 minutes. Pd(dppf)Cl2 (CAS RN72287-26-4) was then added, and the mixture was purged for an additional 15 minutes. The reaction was warmed to 55 °C and stirred at this temperature overnight. The reaction was then diluted with DCM (15 mL), saturated NaHCO3 (20 mL) was added, and washed with brine (20 mL). The organic layer was dried over MgSO4, concentrated under reduced pressure, and purified by silica gel chromatography using 0-60% EtOAc in cyclohexane to give the title compound (97.0 mg, 0.20 mmol, 58%). 28 H 34 LC-HRMS (ESI) calculated for N2O5Na[M+Na]+: 501.2360; found: 501.2388.

[0145] BB11 tert-Butyl 6-(3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylate [ka] Step d Benzyl (2-(3-bromophenoxy)ethyl)carbamate [ka] Analogously to the procedure described for BB10, step d), 3-bromophenol (CAS RN591-20-8) was reacted with 2-(((benzyloxy)carbonyl)amino)ethyl 4-methylbenzenesulfonate (CAS RN 93407-96-6) to give the title compound (366 mg, 0.69 mmol, 69%). LC-MS [M+Na] + :372.0.

[0146] Step 1: tert-Butyl 6-((3-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylate In a similar manner to the procedure described for BB10, step h), benzyl (2-(3-bromophenoxy)ethyl)carbamate was reacted with tert-butyl 6-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylate to give the title compound (181 mg, 0.38 mmol, 63%). 1 H NMR(300MHz,CDCl3)δ 7.41-7.30(m,5H),7.21(t,J=8.1Hz,1H),6.83-6.60(m,3H),6.11(s,1H),5.22(s,1H),5.11(s,2H), 4.08-3.90(m,6H),3.60(d,J=5.5Hz,2H),3.20(s,2H),3.04(s,2H),1.42(s,9H).LC-MS(ESI):[M+H] + :479.2.

[0147] BB12 tert-Butyl 6-(2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylate [ka] Step d Benzyl (5-(2-bromophenoxy)pentyl)carbamate [ka] In a similar manner to the procedure described for BB10, step d), 3-bromophenol (CAS RN 591-20-8) was reacted with 5-(((benzyloxy)carbonyl)amino)pentyl 4-methylbenzenesulfonate (CAS RN 93066-51-4) to give the title compound (403 mg, 1.03 mmol, 51%). 1 H NMR(300MHz,CDCl3)δ 7.52(dd,J=7.9,1.6Hz,1H),7.39-7.20(m,6H),6.94-6.71(m,2H),5.10(s,2H,OCH2),4.78(s,1H,NHCbz),4.0 1(t,J=6.2Hz,2H,OCH2),3.24(q,J=6.3Hz,2H,CH2NHCbz),1.86(t,J=6.8Hz,2H,CH2 linker),1.76-1.48(m,4H,2 x CH2 linker). 13 C NMR(75MHz,CDCl3)δ 156.4,155.3(C=O Cbz,C Ar -O), 136.6, 133.3, 128.5, 128.4, 128.1, 121.8, 113.2, 112.3, 68.8 (OCH2), 66.7 (OCH2), 41.0 (CH2NHCbz), 29.7 (CH2 linker), 28.7 (CH2 linker), 23.3 (CH2 linker).C 19 H 23 BrNO3[M+H] + Calculated for LC-HRMS (ESI): 392.0856; Found: 392.0860.

[0148] Step 1: tert-Butyl 6-(2-((5-(((benzyloxy)carbonyl)amino)pentyl)oxy)benzylidene)-2-azaspiro[3.3]heptane-2-carboxylate In a similar manner to the procedure described for BB10, step h), benzyl (5-(2-bromophenoxy)pentyl)carbamate was reacted with tert-butyl 6-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)-2-azaspiro[3.3]heptane-2-carboxylate to give the title compound (100 mg, 0.19 mmol, 25%). 31 H 41 N2O5[M+H] + Calculated for LC-HRMS (ESI): 521.3010; Found: 521.3000.

[0149] BB14 tert-Butyl (1R,5S)-6-(((2-(2-(((benzyloxy)carbonyl)amino)ethoxy)benzyl)oxy)methyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate [ka] In a similar manner to the procedure described in Example 1g), benzyl (2-(2-(bromomethyl)phenoxy)ethyl)carbamate (see Example 1) was reacted with tert-butyl (1R,5S,6r)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (CAS RN 827599-21-3) to give the title compound (32.3 mg, 0.069 mmol, 57%). 1 H NMR(300MHz,CDCl3)δ 7.40-7.20(m,7H),6.96(td,J=7.5,1.1Hz,1H),6.85(d,J=8.3Hz,1H),5.09(s,2H,-OCH2-),4.52 (s,2H,-OCH2-),4.10(t,J=5.1Hz,2H),3.65-3.39(m,5H),3.36-3.19(m,3H),1.49-1.32(m,11H,3 x CH3Boc,2 x CH fusion cycle),0.98-0.87(m,1H,CH propyl cycle). 13C NMR(75MHz,CDCl3)δ 156.57,156.53,155.0(C=O Cbz,C=O Boc,C Ar -O), 136.6, 129.8, 129.2, 128.6, 128.30, 128.25, 127.0, 121.2, 112.2 (C-Ar), 79.4, 71.7, 68.0, 67.6, 66.9, 48.1 (CH-cyclopropyl), 40.7 (CH-NHCbz), 28.6 (3 x CH-Boc), 27.0, 22.4 (CH-fusion cycle). 28 H 37 N2O6[M+H] + Calculated for LC-HRMS (ESI): 497.2646; Found: 497.2658.

[0150] BB16 tert-Butyl 3-(3-(ethoxycarbonyl)phenoxy)azetidine-1-carboxylate [ka] In a similar manner to the procedure described in Example 39d), ethyl 3-hydroxybenzoate (CAS RN 7781-98-8) was reacted with tert-butyl 3-iodoazetidine-1-carboxylate (CAS RN 254454-54-1) to give the title compound. LC-MS (ESI) [M+Na] + :344.1.

[0151] BB17 tert-Butyl 3-((2-(ethoxycarbonyl)phenoxy)methyl)azetidine-1-carboxylate [ka] In a similar manner to the procedure described in Example 39d), ethyl 3-hydroxybenzoate (CAS RN 7781-98-8) was reacted with tert-butyl 3-(bromomethyl)azetidine-1-carboxylate (CAS RN 253176-93-1) to give the title compound. LC-MS (ESI) [M+Na]+ :358.1.

Claims

1. Formula (I) or (II): 【Chemical 1】 【Chemistry 2】 (In the formula, In formula (I), R １ teeth, 【Chemistry 3】 where the wavy line indicates the R １ (indicating the point of attachment) Selected from: In formula (II), R １ teeth, 【Chemistry 4】 where the wavy line indicates the R １ (indicating the point of attachment) Selected from: R ２ teeth, 【Chemistry 5】 (In the formula: The wavy line indicates the R ２ indicates the point of attachment of p is 1 or 2; V is selected from hydrogen and methyl; W is methyl, phenyl, 【Chemistry 6】 (Wherein, Ar is C ６ -C １０ aryl or 5-14 membered heteroaryl) Selected from: X is O, S, Si(CH ３ ) ２ , and C.H. ２ Selected from: Y is OH, NH ２ , N(CH ３ ) ２ , N(CD ３ ) ２ and azetidin-1-yl Selected from: R ３ is hydrogen, C １ -C ６ Alkoxycarbonyl, and C １ -C ６ Alkoxycarbonyl-NH-C １ -C ６ alkyl-NH-C(O)-; A is, (i) 【Chemistry 7】 and L is -CH ２ -, -CH ２ O-, -OCH ２ -, -CH ２ OCH ２ -, and -O-; or (ii) 【Chemistry 8】 (In the formula, (i) X is CH; L is -CH ２ -, -CH ２ O-, -OCH ２ -, -CH ２ OCH ２ -, and -O-, or (ii) X and L together represent the following group: 【Chemistry 9】 wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I) or (II), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forms and where the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I) or (II); n is an integer selected from 1, 2, 3, and 4. or a pharmaceutically acceptable salt thereof.

2. R １ but, 【Chemistry 10】 where the wavy line indicates the R １ (indicating the point of attachment) 2. A compound of formula (I) or (II) according to claim 1, selected from: or a pharmaceutically acceptable salt thereof.

3. A is, (i) 【Chemistry 11】 and L is -CH ２ O-, -OCH ２ -, -CH ２ OCH ２ -, and -O-; or (ii) 【Chemistry 12】 (In the formula, (i) X is CH; L is -CH ２ - and -CH ２ O-; or (ii) X and L together represent the following group: 【Chemistry 13】 wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forms and wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I).

3. A compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

4. A is, (i) 【Chemistry 14】 and L is -CH ２ OCH ２ -or (ii) 【Chemistry 15】 (In the formula, (i) X is CH; L is -CH ２ - or (ii) X and L together represent the following group: 【Chemistry 16】 wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forms and wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I).

4. A compound of formula (I) according to claim 3, or a pharmaceutically acceptable salt thereof.

5. A is, (i) 【Chemistry 17】 and L is -CH ２ OCH ２ -or (ii) 【Chemistry 18】 wherein X is CH. and L is -CH ２ - and; wherein the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I).

5. A compound of formula (I) according to claim 4, or a pharmaceutically acceptable salt thereof.

6. R ２ but, 【Chemistry 19】 (In the formula, The wavy line indicates the R ２ indicates the point of attachment of p is 1; V is methyl; W is methyl; X is O and Si(CH ３ ) ２ Selected from: Y is N(CH ３ ) ２ is) Selected from: n is an integer selected from 1, 2, and 4; A compound of formula (I) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof.

7. R ２ but, 【Chemistry 20】 (In the formula, The wavy line indicates the R ２ indicates the point of attachment of p is 1; V is methyl; W is methyl; X is O and Si(CH ３ ) ２ Selected from: Y is N(CH ３ ) ２ is) Selected from: n is an integer selected from 1, 2, and 4; 7. A compound of formula (I) according to claim 6, or a pharmaceutically acceptable salt thereof.

8. R ２ but, 【Chemical 21】 (In the formula, The wavy line indicates the R ２ indicates the point of attachment of p is 1; V is methyl; W is methyl; X is O and Si(CH ３ ) ２ Selected from: Y is N(CH ３ ) ２ is) Selected from: n is an integer selected from 1 and 4; 8. A compound of formula (I) according to claim 7, or a pharmaceutically acceptable salt thereof.

9. A is, (i) 【Chemical 22】 and L is -OCH ２ -, -CH ２ OCH ２ -, and -O-; or (ii) 【Chemical 23】 wherein X is CH. and L is -CH ２ is O-; where the wavy line indicates the point of attachment to L and the asterisk indicates the point of attachment to the carbonyl group of formula (II).

3. A compound of formula (II) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

10. R １ but, 【Chemistry 24】 where the wavy line indicates the R １ (indicating the point of attachment) Selected from: R ２ but, 【Chemistry 25】 (In the formula, The wavy line indicates the R ２ indicates the point of attachment of p is 1; V is methyl; W is methyl; X is O and Si(CH ３ ) ２ Selected from: Y is N(CH ３ ) ２ is) Selected from: A is, (i) 【Chemical 26】 and L is -CH ２ O-, -OCH ２ -, -CH ２ OCH ２ -, and -O-; or (ii) 【Chemical 27】 (In the formula, (i) X is CH; L is -CH ２ - and -CH ２ O-; or (ii) X and L together represent the following group: 【Chemical Formula 28】 wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forms and where the wavy line indicates the point of attachment to L and the asterisk indicates the point of attachment to the carbonyl group of formula (I); n is an integer selected from 1, 2, and 4; A compound of formula (I) according to claim 1.

11. R １ but, 【Chemical 29】 where the wavy line indicates the R １ (indicating the point of attachment) Selected from: R ２ but, 【Chemistry 30】 (In the formula, The wavy line indicates the R ２ indicates the point of attachment of p is 1; V is methyl; W is methyl; X is O and Si(CH ３ ) ２ Selected from: Y is N(CH ３ ) ２ is) Selected from: A is, (i) 【Chemical 31】 and L is -CH ２ OCH ２ -or (ii) 【Chemical 32】 (In the formula, (i) X is CH; L is -CH ２ - or (ii) X and L together represent the following group: 【Chemical 33】 wherein the wavy line indicates the point of attachment of L to the phenyl moiety in formula (I), and the two asterisks indicate the points of attachment of X to the respective adjacent atoms in ring A. (forms and where the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I); n is an integer selected from 1, 2, and 4; A compound of formula (I) according to claim 10.

12. R １ but, 【Chemical 34】 where the wavy line indicates the R １ (indicating the point of attachment) Selected from: R ２ but, 【Chemistry 35】 (In the formula, The wavy line indicates the R ２ indicates the point of attachment of p is 1; V is methyl; W is methyl; X is O and Si(CH ３ ) ２ Selected from: Y is N(CH ３ ) ２ is) Selected from: A is, (i) 【Chemical 36】 and L is -CH ２ OCH ２ -or (ii) 【Chemical 37】 wherein X is CH. and L is -CH ２ - and; where the wavy line indicates the point of attachment of A to L and the asterisk indicates the point of attachment of A to the carbonyl group of formula (I); n is an integer selected from 1 and 4; 12. A compound of formula (I) according to claim 11.

13. The compound of formula (I) or (II) benzyl(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.5]nonan-7-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-(((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(3-(3-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamate; benzyl(6-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)hexyl)carbamate; benzyl(2-(2-(((1-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; Cyclohex-1-en-1-ylmethyl(2-(2-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(3-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((7-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-7-azaspiro[3.5]nonan-2-yl)oxy)methyl)phenoxy)ethyl)carbamate; benzyl(5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-ylidene)methyl)phenoxy)pentyl)carbamate; benzyl(2-(2-((((1R,5S)-3-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-3-azabicyclo[3.1.0]hexan-6-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl (R)-(2-(2-(((1-(3-(2-oxooxazolidin-4-yl)propanoyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; benzyl(2-(2-(((1-(3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamate; 2-(6-(2-(2-((7-nitro-1l2,3l2-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one; (4aR,8aS)-6-(3-(((2-(2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; 2-(3-(((2-(2-((7-nitro-1H-312-benzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)-7-oxa-2,5-diazaspiro[3.4]octan-6-one; (4aR,8aS)-6-(3-(((2-(3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)propoxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(3-(((2-((5-((7-nitro-1,3-dihydrobenzo[c][1,2,5]oxadiazol-4-yl)amino)pentyl)oxy)benzyl)oxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(2-(2-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(3-(2-((7-nitro-1,3-dihydrobenzo[c][1,2,5]oxadiazol-4-yl)amino)ethoxy)benzyl)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; 2-(7-(dimethylamino)-3-(dimethyliminio)-5,5-dimethyl-3,5-dihydrodibenzo[b,e]silin-10-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 3,3-dimethyl-1-(6-oxo-6-((2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium 2,2,2-trifluoroacetate; 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborin-3-yl)-N-(2-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)propanamide; 2,2,2-trifluoroacetic acid, 3,3-dimethyl-1-(6-oxo-6-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium salt; 3,3-dimethyl-1-(6-oxo-6-((2-(2-((2-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)ethyl)amino)hexyl)-2-((E)-3-((Z)-1,3,3-trimethylindolin-2-ylidene)prop-1-en-1-yl)-3H-indol-1-ium 2,2,2-trifluoroacetate; 3-(5,5-difluoro-7,9-dimethyl-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborin-3-yl)-N-(2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)propanamide; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((2-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)ethyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((3-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)propyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate compounds and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-((2-(6-oxo-7-oxa-2,5-diazaspiro[3.4]octane-2-carbonyl)-2-azaspiro[3.3]heptan-6-yl)methyl)phenoxy)pentyl)carbamoyl)benzoate; 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-4-((5-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)pentyl)carbamoyl)benzoate and 2-(6-(dimethylamino)-3-(dimethyliminio)-3H-xanthen-9-yl)-5-((5-(2-(((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)methyl)phenoxy)pentyl)carbamoyl)benzoate; Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate; Ethyl 3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzoate; Ethyl 2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzoate; tert-butyl(2-(3-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)oxy)benzamido)ethyl)carbamate; tert-butyl(2-(2-((1-((4aR,8aS)-3-oxooctahydro-2H-pyrido[4,3-b][1,4]oxazine-6-carbonyl)azetidin-3-yl)methoxy)benzamido)ethyl)carbamate; (4aR,8aS)-6-(3-((benzyloxy)methyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; (4aR,8aS)-6-(6-(benzyloxy)-2-azaspiro[3.3]heptane-2-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one; and (4aR,8aS)-6-(3-(phenoxymethyl)azetidine-1-carbonyl)hexahydro-2H-pyrido[4,3-b][1,4]oxazin-3(4H)-one 2. A compound of formula (I) or (II) according to claim 1, selected from: or a pharmaceutically acceptable salt thereof.

14. Use of a compound of formula (II) according to any one of claims 1, 2, 9 and 13 in the manufacture of a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13.

15. A process for preparing a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13, comprising the steps of: Amine 4: 【Chemical 38】 (In the formula, A, L, R １ and n is as defined in claim 1. of, (a) with a fluorescent label containing a carboxylic acid moiety in the presence of a coupling reagent and a base; or (b) with a fluorescent label containing an N-hydroxysuccinimide (NHS) activated ester in the presence of a base; or (c) with 4-chloro-7-nitrobenz-2-oxa-1,3-diazole or 4-fluoro-7-nitrobenz-2-oxa-1,3-diazole in the presence of a base; to obtain the compound of formula (I).

16. A compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 when prepared according to the process of claim 15.

17. A compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 for use in a monoacylglycerol lipase (MAGL) occupancy test.

18. A compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 for use in diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals.

19. A compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 for use in generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

20. Use of a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 in a monoacylglycerol lipase (MAGL) occupancy test.

21. Use of a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 in diagnostic imaging of monoacylglycerol lipase (MAGL) in mammals.

22. Use of a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13 for generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL).

23. A method for testing monoacylglycerol lipase (MAGL) occupancy, comprising contacting MAGL with a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13.

24. A method for diagnostic imaging of monoacylglycerol lipase (MAGL) in a mammal, comprising contacting said MAGL with a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13.

25. 14. A method for generating equilibrium and kinetic binding data for monoacylglycerol lipase (MAGL), comprising contacting MAGL with a compound of formula (I) according to any one of claims 1 to 8 and 10 to 13.

26. 10. The invention as hereinbefore described.

Images