HK1151511B - Preparation and use of alkylating agents - Google Patents

Description

Preparation and use of alkylating agents

The application is a divisional application of Chinese invention patent application (application date: 26/7/2004; application number: 200480027763.X (international application number: PCT/US 2004/024316); invention name: preparation and application of alkylating agent).

Cross Reference to Related Applications

This application claims priority from pending U.S. patent application 60/490,233 filed 24/7/2003 according to 35u.s.c.119(e), which is specifically incorporated by reference.

Technical Field

The present invention relates generally to organic chemistry, and more particularly to methods of synthesizing alkylating agents and methods of use.

Background

Alkylation methods have been used to synthesize and modify a myriad of compounds (see, U.S. Pat. nos. 6277982, 6278020, 6281380, 6281399, 6281405, 6288233, 6291716, 6291724, 6294499, 6303840, 6307048, 6313362, 6315964, 6339179, 6355839, 6376729, 6376730, 6387705, 6388157, 6392114, 6395871, 6395945, 6423871, 6429349, 6440886, 6448458, 6479721, 6486374, 6492571, 6500998, 6504071, 6512153, 6515169, 6525234, 6528316, 6541655, 6548113, 6552241, 6555722, 6642425, 6642426, 6664432, 6673977, 6677269, 6709638, 6747152, 6750354, 6759349). These processes typically require multi-step synthesis and purification of reaction intermediates and/or removal of unwanted by-products. Thus, these methods may be unsuitable or have limited use for synthesizing or modifying radiolabeled compounds, particularly when the radioisotope lifetime is relatively short and the emission-labeled compound is intended for use as a radiotracer or imaging agent (imaging agent). Thus, there is a need in the art for a rapid and efficient alkylation method that can be used to synthesize or modify radiolabeled compounds.

Thus, the present invention provides compositions and methods for the rapid and efficient alkylation of target compounds, which may be used in the synthesis or modification of radiopharmaceuticals.

Disclosure of Invention

The present invention provides a process for alkylating a target compound comprising one or more alkyl reactive groups. In some embodiments, the method comprises synthesizing an alkylating agent. In some embodiments, the alkylating agent may include one or more leaving groups (leaving groups) and an alkylation moiety (alkylation moiety). In some optional embodiments, the alkylating agent may comprise a detectable moiety (detectable moiety). In ethylene embodiments, the alkylating moiety may comprise a detectable moiety. In some embodiments, the detectable moiety may be a radioisotope.

In some embodiments, the alkylating agent may be used in the alkylation reaction without intervening purification steps. Thus, in some embodiments, the alkylating agent, once synthesized, can directly alkylate the target compound. In some embodiments, the synthesis of the alkylating agent and the alkylation of the target compound may occur in a single vessel.

In some embodiments, various aspects of the disclosed methods can be automated by general or special purpose devices. In some embodiments, the apparatus may include a processor or computer that stores data and/or executes computer program code instructions. Thus, in some embodiments, a computer-readable memory that instructs a computer to function in a particular manner may be utilized. A computer or processor may control any one or more aspects of the disclosed methods.

In other aspects of the invention, alkylated and/or labeled target compounds are provided. In some embodiments, the target compound may be labeled with a radiodetectable moiety. Thus, in some embodiments, the alkylated and/or labeled target compounds may be used in vivo or in vitro as therapeutic agents, tracers, and/or imaging agents. In some embodiments, the labeled target compound may be used as an imaging agent for positron emission tomography (positron emission tomography).

Drawings

FIG. 1 depicts an alkylating agent [ 2] according to an embodiment¹⁸F]Synthesis of fluoroethane tosylate.

FIG. 2 depicts an alkylating agent [ 2] according to an embodiment¹⁸F]And (3) synthesizing fluoromethane tosylate.

FIG. 3 depicts dimethylethanolamine [ 2] according to one embodiment¹⁸F]And (3) carrying out fluoralkylation.

FIG. 4 depicts a method according to one embodiment¹⁸F]Structure of fluoroalkylated quaternary amines, 3-quinuclidinyl benzilate (MQNB).

FIG. 5 depicts a method according to one embodiment¹⁸F]Fluoroalkyl quaternary amines, neostigmine (neostigmine), acetylcholinesterase inhibitors.

FIG. 6 depicts a method according to one embodiment¹⁸F]Fluoroalkyl quaternary amines, N-methyl-4-phenyl-pyridinium (MPP), neurotoxin.

FIG. 7 depicts the use of [ 2] according to an embodiment¹⁸F]O-fluoralkylation of tyrosine (Tyr) in dimethyl sulfoxide (DMSO).

FIG. 8 depicts O-fluoroalkylation of a carboxylic acid, benzoic acid, according to one embodiment.

FIG. 9 depicts S-fluoroalkylation of benzylthiol (. alpha. -toluenesulphonol) according to one embodiment.

Fig. 10 provides a schematic diagram of a system suitable for automated synthesis of an alkylating agent, alkylation of a target compound (Tc), and purification of the alkylated target compound, according to an embodiment.

FIG. 11 provides a flow diagram for synthesizing a radio-alkylated target compound according to an embodiment. The numbers on the left of the figure represent approximate time points for an exemplary synthesis.

FIG. 12 depicts the structure of a polystyrene 4-dialkylaminopyridinium, polymeric resin containing covalently attached quaternary ammonium salts according to one embodiment, where R is¹¹And R¹²Is a hydrocarbyl group.

FIG. 13 depicts fluoroalkylation of cysteine to produce [ 2]¹⁸F]Labeled methionine and¹⁸F]-linkage of a labelled methionine to the amino terminus (amino terminus) of the peptide.

FIG. 14 shows an HPLC tracer [ 2] using a radioactivity detector (mV vs. min.), according to an embodiment¹⁸F]FECh. The tracer shows a major radioactive peak, in the sample¹⁸F]FECh。

FIG. 15 shows an HPLC tracing [ using an Ultraviolet (UV) detector (absorbance units (AU) versus time ], according to an embodiment¹⁸F]FECh. This trace shows the major peak detectable by UV absorption, dimethylethanolamine. [¹⁸F]FECh cannot be detected by UV absorption.

FIG. 16 shows an HPLC tracing using a radioactivity detector (mV versus min.), according to an embodiment¹⁸F]FCH. The tracer shows a major radioactive peak, in the sample¹⁸F]FCH。

FIG. 17 shows an HPLC tracer [ 2] using a radioactivity detector (mV vs. min.), according to an embodiment¹⁸F]FCH. This trace shows the major peak detectable by UV absorption, dimethylethanolamine. [¹⁸F]FCH cannot be detected by UV absorption.

Detailed Description

The present invention relates to methods of synthesizing alkylating agents and alkylating target compounds. In some embodiments, the alkylating agent may be labeled. In some embodiments, the label may be transferred to the target compound in an alkylation reaction. In some embodiments, the alkylated target compound is used as a tracer molecule, an imaging agent, or a therapeutic agent. Thus, in some embodiments, the labeled alkylated target compound may be a radiopharmaceutical that may be used to determine the metabolic or physiological state of a cell or tissue in vitro or in vivo. Thus, in some embodiments, the invention provides methods of detecting or monitoring a radioactive compound in a tissue or cell. In some embodiments, the radioactive compound being detected or monitored may be used to assess the metabolic or physiological state of a cell or tissue.

In some embodiments, the presently disclosed methods include synthesizing an alkylating agent and an alkylation of a target compound in a single reaction vessel. In some embodiments, the synthesis and alkylation reactions may be a sequential process without intervening purification steps. In some embodiments, the method further comprises isolating or purifying the alkylated target compound.

In some embodiments, one or more aspects of the methods of the invention can be automated. Thus, in various embodiments, the invention provides one or more automated modules (modules), each capable of performing and/or providing reaction conditions suitable for one or more steps of the methods of the invention. In some embodiments, one or more modules may be controlled by a device having a processor that stores data and/or issues instructions.

Herein, "alkylating agent" and grammatical equivalents thereof refer to a compound having a moiety suitable for forming a hydrocarbyl group in an alkylation reaction, which may be transferred to an alkylation reaction center or group of another compound (e.g., a target compound). As used herein, "alkylation," "alkylation," and grammatical equivalents refer to the following reactions: wherein the alkyl group is transferred to the alkylation reaction center or group of the compound by substitution and/or addition. Thus, in some embodiments, a hydrocarbyl group may be transferred to the alkylation reaction center of the target compound to form an alkylated target compound. By "alkylation reactive center", "alkylation reactive group" and equivalents herein is meant at least one atom of a compound that reacts with an alkylating agent in a reaction in which a hydrocarbyl group may be transferred from the alkylating agent to the compound. In some embodiments, "hydrocarbyl" and grammatical equivalents refer herein to a compound of the formula C_nH_(2n+1)Any of the same family monovalent radicals of (a). Thus, in various exemplary embodiments, the hydrocarbyl group may be methyl, ethyl, propyl, isopropyl, C₂H₃Etc., as will be further described below. In some embodiments, the hydrocarbyl group may further comprise a detectable moiety.

In various exemplary embodiments, the alkylation reaction (alkylation reaction) may include, according to SN₁Or SN₂The mechanism by which the alkylation reaction center of the target compound nucleophilically attacks the electron-deficient region of the alkylating agent is well known in the art (see, e.g., Jerry March, Advanced organic chemistry: Reactions, Mechanisms, and Structure.293-871 (4)^th ed.John Wiley&Sons 1992)). Thus, one of ordinary skill in the art will recognize that in some embodiments, the alkylating agent further comprises one or more Leaving Groups (LGs). As used herein, "leaving group" and grammatical equivalents refer to an atom or molecule that is removed from a molecule, such as an organic molecule. In some embodiments, the residual moiety may be covalently bonded to the targetA hydrocarbyl group of a subject compound. Thus, in various exemplary embodiments, a leaving group may be a charged or uncharged atom or group that leaves an atom or molecule that is considered to be the remainder or major portion of the substrate in a particular reaction. The ability of a leaving group to leave an alkylating agent may be a function of the instability of the leaving group. Thus, the leaving group may influence the intrinsic reactivity of the alkylating agent in the alkylation reaction. In some embodiments, the lower the pKa of the conjugate acid of the leaving group, the more stable the leaving group is because, in some embodiments, the leaving group can more readily stabilize the evolving negative charge that can occur in alkylation reactions. Thus, in some embodiments, the leaving group may be a negatively charged atom or molecule. Examples of leaving groups include, but are not limited to, acetate (AcO), p-nitrobenzoate (PNBO), sulfonate (e.g., Mesylate (Mesylate: MsO)), p-Tosylate (Tosylate: TsO), p-bromobenzenesulfonate (Brosylate: BsO), p-nitrobenzenesulfonate (Nosylate: NsO), fluoromethanesulfonate, difluoromethanesulfonate, Triflate (Triflate: TfO and ethanesulfonate), NH Mesylate, Triflate (Nafimate, Nafimate₃Halide esters, halide ions (e.g., I, Br, Cl), and H₂And O. Thus, in various exemplary embodiments, alkylating agents include, but are not limited to: (TsO)₂CH₂、(TsO)CH₃、(TsO)₂C₂H₅、(TsO)₂C₃H₇、(MsO)C₂H₅、(I)₂CH₂、(I)CH₃、(I)₂C₂H₅、(I)₂C₃H₇、(I)C₂H₅、(Br)₂CH₂、(Br)CH₃、(Br)₂C₂H₅、(Br)₂C₃H₇、(Br)C₂H₅And so on.

In some embodiments, the alkylating agent includes one or more labels. In some embodiments, the group or moiety suitable for transfer to the target compound (i.e., the alkylating moiety) includes one or more labels. As used herein, "label," "detectable moiety," and grammatical equivalents refer to any distinguishing feature of a molecule or compound that is suitable for detection or monitoring. Thus, a label may be an intrinsic feature of a compound and/or a moiety covalently or non-covalently attached to a compound. Labels are well known in the art and are selected at the discretion of the practitioner based on methods of synthesizing, using and/or detecting compounds comprising the label. One of ordinary skill in the art will recognize that, in some embodiments, a marker does not substantially interfere with or inhibit the function or activity of a compound comprising the marker. Thus, in some embodiments, the label of the alkylating agent does not substantially interfere with the alkylation of another compound, e.g., the target compound, by the alkylating agent. It is within the ability of one of ordinary skill in the art to determine the severity or extent to which a label may affect the activity or function of a compound to which it may be attached.

In various exemplary embodiments, labels include, but are not limited to, fluorophores (e.g., richard p. haugland, Handbook of fluorescent Probes and Research laboratories sixthedi (Michelle t. z. spence, ed., Molecular Probes, inc.1996), specifically incorporated by reference); ligands (e.g., haptens, biotin); mobility modifiers (e.g., U.S. patents 5470705, 5514543, 6395486, specifically incorporated by reference); coded microbeads (encoded microbeads) (e.g., U.S. patent nos. 6630307, 6500622, 6274323, expressly incorporated by reference); an enzyme label; an organic or inorganic compound; the amount of the radioactive label (e.g.,¹⁸F，¹¹C，¹⁴C，¹²³I，¹²⁴I，¹²⁵I，¹³¹I，⁷⁶br), or a combination thereof. In some embodiments, the radiolabel preferably has a high specific activity, for example, at least about 600mCi/mmol or more, and/or a short range of positions (positron range), for example, from about 2mm to about 5 mm. In some embodiments, the range of positions may be less than about 2 mm. Thus, in some embodiments, the radioactive label may be suitable positron emission for use in a radioimaging techniqueAn emitter, the radiological imaging technique including, but not limited to, Positron Emission Tomography (PET).

In some embodiments, the alkylating agent has the structure of formula I and includes an alkylating moiety, a leaving group, and a label:

X-(CR¹R²)_aCR³R⁴-LG (I)

wherein the content of the first and second substances,

a is an integer of 0 to 3;

R¹、R²、R³and R⁴Independently H, X or a hydrocarbyl group;

x is H, halogen or a label, and

LG is a leaving group.

In some embodiments:

a is 1;

R¹、R²、R³and R⁴Is H;

x is¹⁸F，¹²³I，¹²⁴I，¹²⁵I，¹³¹I or⁷⁶Br; and

LG is a sulfonate.

In some embodiments:

a is 0;

R³and R⁴Is H;

x is [ 2]¹⁸F](ii) a And

LG is tosylate, mesylate or triflate.

In some embodiments:

a is 0;

R³and R⁴Is H or X;

x is [ 2]¹⁸F](ii) a And

LG is tosylate, mesylate or triflate.

Thus, in various exemplary embodiments, alkylating agents comprising a radiolabel include, but are not limited to, haloalkyl sulfonates, such as¹⁸F]Fluorinated alkyl sulfonate (e.g., [ alpha ]¹⁸F]Fluoroethane sulfonate (e.g., and¹⁸F]fluoroethane tosylate [ alpha ], [ alpha¹⁸F]Fluoroethane methanesulfonate [ sic ], [ solution of a salt of a fluoroethane sulfonic acid¹⁸F]Fluoroethane triflate [ f ], [ solution of (b) ]¹⁸F]Fluoromethanesulfonate (e.g., etc.)¹⁸F]Fluoromethane tosylate [ alpha ], [ beta¹⁸F]Fluoromethane methanesulfonate [ alpha ], [ alpha¹⁸F]Fluoromethane triflate)), (a) a⁷⁶B]Brominated hydrocarbyl sulfonate (e.g., etc.)⁷⁶B]Bromoethane sulfonate (e.g., etc.)⁷⁶B]Bromoethane tosylate [ alpha ], [ beta⁷⁶B]Bromoethane methanesulfonate [ alpha ], [ beta ]⁷⁶B]Bromoethane trifluoromethanesulfonate (bromoethane-trifluoromethanesulfonate), (2)⁷⁶B]Bromomethane sulfonate (e.g., etc.)⁷⁶B]Bromomethane tosylate [ alpha ], [ beta⁷⁶B]Bromomethane methanesulfonate [ alpha ], [ alpha⁷⁶B]Bromomethane trifluoromethanesulfonate)), (a)¹²⁵I]Iodo-hydrocarbyl sulfonate (e.g., etc¹²⁵I]Iodoethane sulfonate (e.g., etc.)¹²⁵I]Iodoethane tosylate [ alpha ], [ beta ]¹²⁵I]Iodo-ethane methanesulfonate, a¹²⁵I]Iodo-ethane triflate (f), and¹²⁵I]iodomethane sulfonate (e.g., etc.)¹²⁵I]Iodomethane tosylate [ alpha ], [ beta ]¹²⁵I]Iodomethane methanesulfonate [ alpha ], [ alpha ]¹²⁵I]Iodomethane triflate), and the like.

In various exemplary embodiments, alkylating agents have been used in methods of alkylating or labeling a target compound. As used herein, "target compound" and grammatical equivalents refer to a compound that contains one or more alkylation reaction centers and, thus, may be alkylated by any one or more alkylating agents, alone or in any combination, by the methods disclosed herein. In some embodiments, two or more target compounds may be alkylated by any one or more alkylating agents simultaneously and/or sequentially. It is within the ability of one of ordinary skill in the art to determine the suitability of any one or more alkylating agents to alkylate one or more target compounds or one or more alkylation reaction centers simultaneously or sequentially. In some embodiments, the compound of interest comprises a label as described above. Thus, in some embodiments, the label does not substantially interfere with or interfere with the alkylation and/or use of the alkylated target compound.

In some embodiments, the alkylation reactive center or group of the target compound may be a suitable nucleophile for the alkylation reaction. Examples of alkylation reaction centers include, but are not limited to, N, O, S, P, C, aldehydes, aliphatic carbons, alkanes, alkenes, alkynes, alcohols (-OH), amines (e.g., primary, secondary, tertiary, and quaternary amines), aromatics (e.g., benzene, phenol), carboxylic acids (-COOH), esters, diazonium ions, dithianes (dithianes), enamines, enolates, heterocycles, hydrazones, imines, ketones, nitriles, oxazines, oxazolines, selenium sulfoxides (selenoxides), sulfones, sulfonates, and cyclic, straight chain, branched chain, substituted, or unsubstituted derivatives thereof. Specifically incorporated by reference as a Dictionary of Chemical terminologies (Parker, et al. eds., McGraw-Hill Book Co.), Encyclopedia of Chemistry (Considine, et al. eds., 4)^thed., VanNostrand Reinhold Co.), March, Advanced Organic Chemistry). In some embodiments, the alkylation reactive group may be hydrocarbyl, substituted hydrocarbyl, saturated and unsaturated cycloalkyl, aryl, substituted aryl, sulfhydryl (-SH), amino, and saturated and unsaturated heterocyclic rings containing, for example, one or more of nitrogen, oxygen, sulfur, and combinations thereof.

As used herein, "alkyl group" refers to a saturated or unsaturated, branched, straight chain or cyclic monovalent hydrocarbon radical derived by the removal of a hydrogen atom or a leaving group from a single carbon atom of a parent alkane, alkene or alkyne. If branched, it may be branched at one or more positions and may be branched at any position unless specified. Typical hydrocarbyl groups include, but are not limited to: a methyl group; an alkyl group such as an ethyl group, a vinyl group, an ethynyl group; a propyl hydrocarbon group such as propan-1-yl, propan-2-yl, cycloprop-1-yl, propan-1-en-2-yl, propan-2-en-1-yl (allyl), cycloprop-1-en-1-yl; prop-2-en-1-yl, prop-1-yn-1-yl, prop-2-yn-1-yl, and the like; butylhydrocarbon groups such as but-1-yl, but-2-yl, 2-methyl-prop-1-yl, 2-methyl-prop-2-yl, cyclobut-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-2-yl, but-1, 3-dien-1-yl, butyl-1, 3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobut-1, 3-dien-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, and the like.

Thus, in some embodiments, "hydrocarbyl" refers to a group having any degree or level of saturation, i.e., a group having only single carbon-carbon bonds, a group having one or more double carbon-carbon bonds, a group having one or more triple carbon-carbon bonds, and a group having a mixture of single, double, and triple carbon-carbon bonds. When it is intended to indicate a specific saturation level, the terms "alkyl", "alkenyl" and "alkynyl" are used. In various exemplary embodiments, the hydrocarbyl group includes from about 1 to about 20 carbon atoms (C)₁-C₂₀Hydrocarbyl group) of about 1 to about 15 carbon atoms (C)₁-C₁₅Hydrocarbyl group) of about 1 to about 10 carbon atoms (C)₁-C₁₀Hydrocarbyl groups), or from about 1 to about 6 carbon atoms (C)₁-C₆Hydrocarbyl groups), or from about 1 to about 3 carbon atoms (C)₁-C₃)。

The term "substituted hydrocarbyl" refers to the above-described hydrocarbyl groups in which one or more hydrogen atoms may be substituted with substituents such as halides (e.g., F, Br, I, Cl), hydrocarbyl groups, amines, aromatics, aryls, hydroxyls, carbonyls, azos, imines, nitriles, nitro groups, sulfhydryl groups, and sulfonyl groups.

The term "cycloalkyl" refers to a group having up to about 20 carbon atoms (C)₂₀) The above hydrocarbon group having a cyclic structure of (1).In various exemplary embodiments, cycloalkyl groups may be monocyclic or polycyclic, e.g., bicyclic. The cycloalkyl group may optionally contain one or more carbon-carbon double bonds, provided that the group is not aromatic. Thus, in various exemplary embodiments, the cycloalkyl group may be saturated or unsaturated.

As used herein, "aryl" refers to a monocyclic or polycyclic aromatic hydrocarbon group (C) containing from about 6 to about 20 carbon atoms₆-C₂₀Aryl), monocyclic or polycyclic aromatic hydrocarbon groups of about 6 to about 15 carbon atoms (C)₆-C₁₅Aryl) or monocyclic or polycyclic aromatic hydrocarbon groups of about 6 to about 10 carbon atoms (C)₆-C₁₀Aryl), and any carbocyclic ketone or thione derivative thereof, wherein the carbon atom having a free valence is a constituent of an aromatic ring (for example, aryl includes phenyl, naphthyl, anthryl, phenanthryl, 1, 2, 3, 4-tetrahydro-5-naphthyl, 1-oxo-1, 2-dihydro-5-naphthyl, 1-thioxo-1, 2-dihydro-5-naphthyl, and the like).

In some embodiments, the aryl group can be heteroaryl. "heteroaryl" refers to monocyclic or polycyclic aromatic hydrocarbon groups containing a total of 6 to 20 atoms, and any heterocyclic ketone and thione derivatives thereof (e.g., thienyl, furyl, pyrrolyl, pyrimidinyl, isoxazolyl, oxazolyl, indolyl, benzo [ b)]Thienyl, isobenzofuranyl, purinyl, isoquinolyl, pteridinyl, pyrimidinyl, imidazolyl, pyridyl, pyrazolyl, pyrazinyl, 4-oxo-1, 2-dihydro-1-naphthyl, 4-thioxo-1, 2-dihydro-1-naphthyl, and the like), wherein at least one to about 5 of the designated carbon atoms are substituted with a heteroatom selected from N, O, S, P or As, wherein the carbon atom having a free valence is part of an aromatic ring. Thus, hetero (C)₆) Aryl includes the groups pyridyl, pyrimidyl, and the like.

In some embodiments, the aryl group may be a substituted aryl group. "substituted aryl" refers to the above-described aryl groups in which one or more hydrogen atoms may be substituted with a substituent, such as, for example, halide (e.g., F, Br, I, Cl), hydrocarbyl, amine, aromatic, aryl, hydroxyl, carbonyl, azo, imine. Nitrile, nitro, sulfhydryl and sulfonyl.

As used herein, "heterocycle" refers to a ring made up of more than one atom. Thus, in some embodiments, heterocyclic rings include, but are not limited to, carbocyclic rings having at least one additional atom in the ring. In various exemplary embodiments, the heterocyclic ring may be saturated or unsaturated. In some embodiments, the heteroatom may be bonded to a ring structure. In some embodiments, the heterocyclic ring may have up to 20 atoms in one or more rings. Thus, in various exemplary embodiments, the heterocycle may be monocyclic or polycyclic, e.g., bicyclic.

Thus, in various exemplary embodiments, the compound of interest may include an organic compound, an inorganic compound, a naturally occurring compound (e.g., a compound isolated from a natural source (e.g., a polypeptide, a nucleic acid, a hormone, a cytokine, an antibody, etc.), a non-naturally occurring compound (e.g., a substance synthesized or known to be not found in nature, such as a nucleic acid Peptide (PNA)), a drug (e.g., an antiviral agent, a pro-drug), and/or combinations thereof. In some embodiments, the target compound may be a biological compound. As used herein, "biological compound", "biochemical compound" and grammatical equivalents refer to a compound that has at least one biological or biochemical activity in vivo (e.g., in a subject) or in vitro (e.g., in tissue culture or in an assay). Thus, in various exemplary embodiments, a biologically active compound can specifically or non-specifically bind to another molecule and/or cell and/or enter a cell of interest. In some embodiments, the cell can be a prokaryotic cell, a eukaryotic cell (e.g., a tumor cell), a microparticle, an anucleated cell (enucleated cell), an enucleated cell (enucleated cell), and the like. In various exemplary embodiments, binding to a molecule (e.g., an antibody or binding portion thereof), a Class of molecules (e.g., MHC Class II molecules), a specific receptor or Class of receptors, and the like can be provided. In some embodiments, the biologically active compound preferably passes through one or more layers of cell membranes and/or cell walls. In some embodiments, the biologically active compound may be accumulated in an organelle or a local (e.g., cytoplasm, mitochondria, endoplasmic reticulum, one or more surfaces of a membrane, etc.). Biologically active compounds can enter cells by various mechanisms, e.g., active or passive mechanisms. These mechanisms include, but are not limited to, diffusion and endocytic mechanisms, e.g., receptor-mediated endocytosis, as well as active and passive transport. In some embodiments, favorable cells take up the biologically active compound at a rate or amplitude that is significantly greater than other cells. Thus, the present invention contemplates biologically active compounds that enter favorable cells at a rate that exceeds other cells and biologically active compounds that accumulate in favorable cells at a final concentration that exceeds the concentration of other cells. For example, in some embodiments, the biological compound preferentially enters tumor cells as compared to non-tumor cells.

In some embodiments, the bioactive compound may be metabolized by the cell upon entering the cell, thereby altering the hydrophobicity of the bioactive compound. In some embodiments, the hydrophobicity of the biologically active compound is reduced upon modification, facilitating retention of the compound in the cell. The hydrophobicity of a biologically active compound can be reduced by extensive cellular treatment, for example, by the addition of polar groups (e.g., hydroxyl groups) or charged groups (e.g., phosphate, carboxylate), by oxidation (e.g., conversion of hydroxyl groups to carbonyl groups), or by removal of hydrophobic groups (e.g., hydrocarbyl groups). Methods by which the hydrophobicity of a biologically active compound can be reduced are within the ability of one of ordinary skill in the art and may be based on the biologically active compound and the metabolic processes of the beneficial cell.

Examples of biologically active compounds include, but are not limited to, choline, dimethylethanolamine (DeGrado, et al (2001) J.Nucl.Med. (42: 1805-1814); U.S. Pat. No. 6630125; Hara, et al (2002) J.Nucl.Med.43 (2): 187-199; Hara, et al. Japanese Patent Office, Patent journal (A); Kokai Patent Application No. HEI 9[1997 ])]-48747), morphine, heroin, pethidine (pethine), tamoxifen, codeine (cod)eine), nicotine, thioproperazine (thioproperazine), diazepam (diazepam), caffeine, flunitrazepam (flunitrazepam), hexamethonium (hexamethonium), methionine (methiodide), quinuclidinyl diphenylglycolate, glucose, deoxyglucose, lactic acid, cyclohexabatal, thymidine (thymidyline), iodoantipyrine (iodoantipyrine), antipyrine (antipyrine), coenzyme Q, adenosylmethionine (adenosylmethionine), aniline (Huang, et al. (2002) nucleic acid media biol.29 (741. 751), CP-126, 998(Musachio, et al (2002) nucleic acid media.29: 547), cocaine derivatives (552et al (2002) nuclear med. biol.29: 19-27), proteins and peptides (us patent 6358489), neurokinin-1 receptor antagonists (us patent 6241964, 6187284), spiroperipherical piperidones (u.s.inv.reg.h1209, us patent 4871527), fatty acids (us patent 6362352), platelet GP IIb/IIIa receptor antagonists (us patent 5879657, 6022523), fluorometholonidazole (fluoromonidazole) (us patent 5886190), tamoxifen derivatives (us patent 5219548), opioid ligands (opioid ligands) (us patent 4775759; ravert, et.al. (2002) nuclear.med.biol.29: 47-53; ogawa, et al (2001) nuclear.med.biol.28: 941 947), non-steroidal aromatics (us 6019957), amino acid analogs (us 5187776, 5808146), MQNB, neostigmine, MPP, NMS, amino acids (e.g., tyrosine (e.g., -OH), serine (e.g., -OH), threonine (e.g., -OH), cysteine (e.g., -S), aspartic acid (e.g., -COOH), glutamic acid (e.g., -COOH), carboxylic acids (e.g., benzoic acid and amino acids), spiperone (spiperone), spiroperidol (spiperodol).

The alkylating agent may be synthesized from a disubstituted alkyl precursor. Thus, in some embodiments, the precursor of the alkylating agent may have the structure of formula II:

LG¹-(CR¹R²)_aCR³R⁴-LG² (II)

wherein:

a is 0 to 2;

R¹、R²、R³and R⁴Independently H, marker, CH₃、C₂H₃(ii) a And

LG¹and LG²Is a leaving group.

In some embodiments:

a is 0 to 1;

R¹、R²、R³and R⁴Is H; and

LG¹and LG²Is a sulfonate or¹⁸F，¹²³I，¹²⁴I，¹²⁵I，¹³¹I，⁷⁶Br。

In some embodiments:

a is 0 to 1;

R¹、R²、R³and R⁴Is H; and

LG¹and LG²Independently a tosylate, mesylate or triflate.

In some embodiments:

a is 1;

R¹、R²、R³and R⁴Is H; and

LG¹and LG²Both are tosylate, mesylate or triflate.

In some embodiments:

a is 0;

R³and R⁴Is H; and

LG¹and LG²Both are tosylate, mesylate or triflate.

Thus, in various exemplary embodiments, precursors of alkylating agents include, without limitation, disulfonates, such as ditriflesulfonylethane, dimethylsulfonylethane, bis (trifluoromethanesulfonyl) ethane (ditriflymethane), ditriflesulfonylmethane, dimethylsulfonylmethane, and bis (trifluoromethanesulfonyl) methane (ditriflymethane).

In some embodiments, the alkylating agent may be synthesized by halogenating a precursor thereof. In some embodiments, the halogen may be a radioisotope (e.g.,¹⁸F，¹²³I，¹²⁴I，¹²⁵I，¹³¹I，⁷⁶br) and thus may also be a label. In a preferred embodiment, the halogen may be¹⁸F. In some embodiments of the present invention, the substrate is,¹⁸f can be [ 2]¹⁸F]Fluorides and can be prepared by a number of any methods known in the art. In some embodiments, the production may be by irradiation with high energy protons in a cyclotron¹⁸F to obtain the product¹⁸F]HF/[¹⁸O]H₂O, as is known in the art 400. In some embodiments, the [ alpha ], [ beta¹⁸F]Fluoride may be purified by chromatography, including but not limited to ion exchange chromatography 410 and other methods, as known in the art.

Methods for producing other radioisotopes are well known in the art (see, e.g., Handbook of radiopharmaceuticalica (Welch and Radvanly, eds., John Wiley& Sons，&2003) And Chemists' Views of Imaging Centers291-295(Emran, ed., Penum Press.1995)). For example, methods for producing iodine radionuclides can be found in the following documents: chemistry Applied to Iodine raditions of Finn, Handbook of Radiopharmaceuticals423-400(Welch and Redvanly, eds., John Wiley& Sons，2003) And Production and Application of Washburn et al¹²³I-LabeledM-Iodobetizylguanidine(¹²³I-MIBG), Chemists' Views of Imaging Centers291-295(Emran, ed., Penum Press 1995). Methods for producing bromine radionuclides can be found in Radiobromine for Imaging and Therapy 441-&Sons，2003) Is found in (1).

In some embodiments, the method may be as described in Block et al, 1987, j.label.compds.radiopharman.24: 1029-41. However, in some embodiments, the temperature of the halogenation reaction may be important in determining the yield of the alkylating agent. Thus, in various exemplary embodiments, the temperature of the halogenation reaction may range from about 40 ℃ to less than 80 ℃, from about 40 ℃ to about 50 ℃, from about 45 ℃ to about 50 ℃, or from about 20 ℃ to about 50 ℃. Generally, without being bound by theory, when halogenating the alkylating agent precursor, the synthesized alkylating agent is more reactive with halogen than the precursor, which results in the production of byproducts including, but not limited to, dihaloalkanes. Thus, lower temperatures (e.g., less than about 50 ℃) may reduce the production of byproducts. One of ordinary skill in the art will recognize that lower temperatures decrease the rate of halogenation. Thus, in some embodiments where the halogen is a radioisotope, the reaction temperature and rate may be adjusted such that the extent of radioisotope decay during the reaction is minimized. It is within the ability of one of ordinary skill in the art to determine the optimum reaction temperature and rate ranges for the halogenation reaction. However, in some embodiments, reaction temperatures less than about 40 ℃ may be unsuitable when the half-life of the radioisotope is less than about 2 hours. In some embodiments, the halogenation reaction may occur in the presence of a catalyst that catalyzesAgents include, but are not limited to(for example,2.2.2) or a basic tetraalkylammonium salt (e.g., tetrabutylammonium bicarbonate) 410. In some embodiments, the potassium carbonate can be a halogen ion such as [ sic ], [ solution ]¹⁸F]The fluoride provides the counter ion. However, the choice of the source and type of counter ion is within the ability of one of ordinary skill in the art and may be influenced by the choice of the type of halide ion and/or target compound at the discretion of the practitioner. In some embodiments, the halogenation reaction may be stirred by any means including, but not limited to, shaking, mixing, and bubbling a substantially inert gas (e.g., argon, nitrogen, helium, etc.) into the reaction 450. In some embodiments, stirring may be maintained as long as possible during the reaction.

In various exemplary embodiments, a solid support or resin may be used to minimize or prevent the synthesis of dihalohydrocarbyl byproducts. In some embodiments, the solid support or resin is bound to a halide ion that can be used for the halogenation precursor. In some embodiments, the alkylating agent precursor may be a disulfonate ester, as described above. In some embodiments the resin may be a polymeric resin, e.g., polystyrene, containing covalently attached quaternary ammonium salts (e.g., quaternary 4-dihydrocarbylaminopyridinium salts) (fig. 12). In some embodiments, the solid support may be QMA.

In some embodiments, the purified halide ion, e.g., [ solution ]¹⁸F]The fluoride may be dried using methods known in the art including, but not limited to, azeotropic drying 420 and cooling 430 under a flow of argon at a temperature of at least about 110 ℃ to at least about 115 ℃. In some embodiments, the azeotropic drying temperature may be above or below this range, however, one of ordinary skill in the art will recognize the suitability of drying the halide ion for synthesizing the alkylating agent and the downstream alkylating agentThe effect of use is not the effect of the actual temperature at which drying is carried out.

In alternative embodiments, the alkylating agent, once prepared, may be used, with or without purification, to alkylate or label target compound 460. "purification", "purifying" and grammatical equivalents herein refer to the reduction of the amount of foreign material, e.g., by-products, from a beneficial material. Thus, in some embodiments, the alkylating agent, once prepared, may be used directly to carry out alkylation or to label the target compound. In some embodiments, the target compound may be added directly to the vessel used to synthesize the alkylating agent. Thus, in some embodiments the alkylating agent and the alkylated target compound may be prepared in a coupled reaction (coupled reaction), which in some embodiments may be a "one pot, two step" process.

In some embodiments, the target compound may be liquid at the beginning of the alkylation reaction, and the amount may be from about 0.2mL to about 0.5 mL. In some embodiments, the target compound may be used neat. In some embodiments, the target compound may be diluted or dissolved in a solvent including, but not limited to, acetonitrile, Dimethylsulfoxide (DMSO), Dimethylformamide (DMF), and Tetrahydrofuran (THF). The choice of the type of solvent to use with the target compound is within the ability of one of ordinary skill in the art. In various exemplary embodiments, alkylation of the target compound may be carried out at about 100 ℃, about 100 ℃ to about 130 ℃, or about 100 ℃ to about 110 ℃, or about 100 ℃ to about 105 ℃ for about 10 minutes to about 25 minutes, and may be stirred as described above at 460. The choice of temperature and reaction time is within the ability of one of ordinary skill in the art.

In some embodiments, the target compound is alkylated according to scheme 1:

scheme 1

LG-(CH₂)_nR⁵+Q_mZ→Q_mZ(CH₂)_nR⁵

Wherein the content of the first and second substances,

n is an integer of 1 to 3;

R⁵is H or a marker (X);

LG is a leaving group;

Q_mz is the alkylation reaction center of the target compound, wherein m is an integer from 1 to 4, each Q is independently H, hydrocarbyl, substituted hydrocarbyl, cycloalkyl, aryl, and substituted aryl, and wherein Z is N, O, S, P, or C.

In some embodiments of the present invention, the substrate is,

n is an integer of 1 to 2;

R⁵is a label (X);

LG is a sulfonate;

Q_mz is the alkylation reaction center of the target compound, wherein m is an integer from 1 to 4, each Q is independently H, hydrocarbyl, substituted hydrocarbyl, cycloalkyl, aryl, and substituted aryl, and wherein Z is N, O, S, P, or C.

In some embodiments of the present invention, the substrate is,

n is an integer of 1 to 2;

x is a radioactive label;

LG is mesylate, tosylate or triflate;

Q_mz is an alkylation reaction center of the target compound, wherein m is an integer of 1 to 3, and each Q_1-3Independently methyl, hydroxyethyl, and wherein Z is N.

In some embodiments, when the target compound is alkylated or labeled, it can be purified by various methods known in the art. For example, the alkylated target compound may be purified by electrophoresis, precipitation, extraction, and/or column chromatography 470 (e.g., ion exchange chromatography, molecular exclusion chromatography). It is within the ability of one of ordinary skill in the art to identify any one or more purification steps or methods for purifying the alkylated or labeled target compound. Thus, one of ordinary skill in the art will recognize that purification of the alkylated or labeled compound by, for example, chromatography may include one or more washing steps 480, 490 prior to elution 500. In some embodiments, the pH or ionic strength 510, 510 of the alkylated or labeled compound may be later altered. In some embodiments, the alkylated or labeled compound may be purified such that it is suitable for use in living body 520.

By way of example, and not limitation, the target compound for alkylation may be N-fluoromethyl-MQNB (FIG. 4), N-fluoromethyl-MPP (FIG. 6), N-fluoromethyl spiroperone (FNMS), [ 2]¹⁸F]Fluoromethyl-neostigmine, and¹⁸F]fluoromethyl-tyrosine and 3- (2' -fluoroethyl) spiperone (FESP). In some embodiments, the alkylated target compound may also be labeled. Thus, in some embodiments, the present invention contemplates radiolabeled target compounds. Thus, in various exemplary embodiments, fluorine in each of the above compounds may be substituted with one or more fluorine atoms¹⁸F，¹²³I，¹²⁴I，¹²⁵I，¹³¹I, or⁷⁶Br is substituted.

In some embodiments, the alkylated target compound may have a structure of formula III:

wherein the content of the first and second substances,

r' is¹⁸F，¹²³I，¹²⁴I，¹²⁵I，¹³¹I or⁷⁶Br；

B-is a counterion;

b is an integer of 1 to 3;

R⁶、R⁷、R¹⁰independently is CH₃，C₂H₅，C₃H₇，C₆H₅；

R⁸、R⁹Independently is H, CH₃，C₂H₅。

Thus, in various exemplary embodiments, the target compound for alkylation may be a choline analog, e.g., N, N-dimethyl-N-fluoromethylethanolamine (fluorocholine: FCH), N, N-dimethyl-N-fluoroethylethanolamine (fluoroethylcholine: FECh), etc¹⁸F]FCH, and [ 2]¹⁸F]FECh, N, N-dimethyl-N-bromomethylethanolamine (bromocholine: BrCH), N, N-dimethyl-N-bromoethylethanolamine (bromoethylcholine: BrECh), and⁷⁶Br]BrCH and [ 2]⁷⁶Br]BrECh, N, N-dimethyl-N-iodomethylethanolamine (iodocholine: ICH), N, N-dimethyl-N-iodoethylethanolamine (iodoethylcholine: IECh), N, N-dimethyl-N-iodomethylethanolamine, N, N-dimethyl-N-iodoethylethanolamine, N, N-dimethyl-¹²⁵I]ICH, and [ solution ]¹²⁵I]IECh, etc.

In some alternative embodiments, the alkylated target compound may be different from [ 2]¹⁸F]FCH，[¹⁸F]FECh or [ 2]¹⁸F]FDG (fluorodeoxyglucose), for example, wherein the alkylating agent used to prepare the alkylated compound is purified prior to the alkylation reaction.

In some embodiments, the preparation¹⁸F]The method of labeling the target compound (Tc) may comprise [ alpha ]¹⁸F]Fluoride and X- (CH)₂)_n-X, to form [ 2]¹⁸F]-(CH₂)_n-X, wherein X is tosylate, mesylate or triflate, and n ═ 1-4, adding a target compound comprising an alkylation reactive group to the reaction vessel to form [, [ solution ], or [ solution ], or¹⁸F]-(CH₂)_n-T_cPassing the reaction mixture through a first chromatographic support to bind the [ 2]¹⁸F]-(CH₂)_n-T_cAnd eluted from the carrier¹⁸F]-(CH₂)_n-T_c. In some embodiments, the reaction vessel may be in fluid communication with a first chromatographic carrier. In some embodiments, the first chromatography carrier has an outlet in fluid communication with a dispenser for aliquoting the chromatography carrier¹⁸F]-(CH₂)_n-T_cThe eluate was transferred to a single vial. In some embodiments, the second chromatography carrier may be in fluid communication with and located between the first chromatography carrier and the dispenser.

Once prepared, the alkylated or labeled compounds (derivatives of the subject compounds) can be used in vivo or in vitro at the discretion of those of ordinary skill in the art. In some embodiments, the second target compound may be modified or labeled using an alkylated or labeled compound. As shown in fig. 13, labeled amino acids can be bound to the peptide. In some embodiments, the binding method may employ solid phase peptide synthesis as described in, for example, Bianco et al, 2003, org.biomol.chem.1 (23): 4141-3(Epub 2003 Oct 22); deechongkit et al, 2004, Org Lett.6 (4): 497-500; hojo et al, 2004, chem. pharm. ball. (Tokyo)52 (4): 422-7; malkinson et al, org.lett.5 (26): 5051-4; merrifield, 1995, Biopolymers 37 (1): 3-4; merrifield, 1997, Methods Enzymol.289: 3-13; song, et al, 2004, bioorg.med.chem.lett.14 (1): 161-5; stephenson et al, 2004, bioconjugate. chem.15 (1): 128-36; wang et al, 1987, int.J.Pept.protein Res.30 (5): 662-; and us patents 4507230, 4816513, 5186898, 5233044, 5258454, 5286846, 5380495, 5444150.

In some embodiments, the labeled target compound may serve as a therapeutic agent, an imaging agent, a radiopharmaceutical agent, or a tracer. Thus, in some embodiments, alkylated or labeled compounds may be used to detect, monitor or analyze cells or tissues in a subject. In some embodiments, the cells or tissue may be benign, malignant, neoplastic (neoplastic), and/or cancerous, and may be staged for implementation by imaging techniques. In some embodiments, the imaging technique may be Positron Emission Tomography (PET). Methods and techniques for carrying out PET are well known in the art (see us patents 4563582, 4642464, 4647779, 4677299, 4733083, 4864138, 4864140, 5027817, 5103098, 5138165, 5210420, 5224037, 5319204, 5378893, 5453623, 5591977, 5602395, 5744802, 5827499, 5998792, 6130430, 6288399, 6434216, 6521893, 4066783, 6718006). Thus, in some embodiments, alkylated or labeled compounds may be formulated at the discretion of the practitioner according to their intended use. When used as an imaging agent in vivo, the labeled target compound is formulated to be suitable for intravenous injection (e.g., sterile, pyrogen-free). In some embodiments, the labeled compound of interest may be formulated with a pharmaceutically acceptable carrier, such as a pharmaceutically acceptable salt, and administered to the subject in a pharmaceutically effective amount.

By "pharmaceutically acceptable salt" herein is meant a salt of a compound of the invention prepared with a counterion, which is understood in the art to be pharmaceutically acceptable in general and to possess the desired pharmaceutical activity of the parent compound. Such salts include, but are not limited to: (1) acid addition salts with inorganic or organic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, laurylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) a salt formed when an acidic proton present in the parent compound is replaced with a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion); or a salt formed by coordination with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, morpholine, piperidine, dimethylamine, diethylamine, and the like. Also included are salts of amino acids such as arginine, salts of organic acids such as glucuronic acid or galacturonic acid (see Berge et al, 1977, J.pharm.Sci.66: 1-19).

"pharmaceutically effective amount" or "therapeutically effective amount" refers to an amount sufficient to produce a desired physiological effect or to enable a desired effect to be obtained, particularly with respect to diagnosing or treating a condition or disease state, including reducing or eliminating one or more syndromes of the condition or disease, or preventing a disease or state. Thus, in a preferred embodiment, a sufficient amount of a labeled compound of interest is administered to detect, monitor and/or stage a neoplasm in a subject. The amount administered and the route of administration are selected at the discretion of the practitioner, as is known in the art.

Formulations suitable for parenteral administration, for example by the internodal (in the joints), intravenous, intramuscular, intradermal, intraperitoneal and subcutaneous routes, include: aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents, solubilizers, thickeners, stabilizers, and preservatives. In the practice of the present invention, the composition may be administered, for example, by intravenous infusion, orally, topically, intraperitoneally, intravesically, or intrathecally. Parenteral, oral, subcutaneous and intravenous administration are preferred methods of administration.

In various exemplary embodiments, the methods of synthesizing alkylating agents, alkylating or labeling target compounds, and/or purifying alkylated or labeled compounds described above may be performed automatically. Thus, in some embodiments, the methods disclosed herein may be implemented on a general-purpose or special-purpose device, such as a device having a processor that stores data and/or issues commands. It should be appreciated that the computing device may be a single computer or a plurality of networked computers and related to implementing the methods and processes described hereinSeveral programs may be implemented on one or more computer devices. In some embodiments, the disclosed processes and methods are performed on a standard server-user network infrastructure, thereby adding or being compatible with the features disclosed in the various embodiments. The methods, processes, and procedures described herein may generally be performed in software, hardware, and/or combinations thereof. In some embodiments, as shown in fig. 10, the processor 300 may be controlled by a readable memory 310. In some embodiments, the processor 300 can be programmed to instruct the use of QMA 170 to purify a halogen ion such as¹⁸F]Fluoride 100. In some embodiments, the processor 300 instructs elution from QMA 170 using K222/carbonate 110¹⁸F]Fluoride 100, and directing the eluate to a Reaction Vessel (RV) 210. In some embodiments, the xylene sulfonyl methane 120 can be directed to the reaction vessel 210. For the synthesis of alkylating agents, for example, [ 2]¹⁸F]Fluoromethane tosylate, processor 300 may instruct temperature control module 320 to heat reaction vessel 210 to a programmed temperature (programmed temperature) selected at the discretion of the practitioner, and may control the mixing of reactants by bubbling an inert gas such as argon 180. In some embodiments, processor 300 instructs the addition of dimethylamine 130 to RV 210, RV 210 can be heated by thermal control die 320 and mixed by argon 180 to synthesize an alkylated or labeled compound, e.g., [ solution ], [¹⁸F]FCH. EtOH/water 140 may be introduced to reaction vessel 320 by processor 300, which EtOH/water 140 in some embodiments washes reaction vessel 320 and obtains additional alkylation product. In some embodiments, processor 300 introduces the contents of reaction vessel 320 to silica column 200. The waste 230 is not bound to the silica column 200, and in some embodiments, the alkylation product is, for example, [ 2]¹⁸F]FCH may be eluted by AcOH 150. The AcOH 150, alkylation product, can be removed by introducing the eluate into a weakly basic ion exchange resin 240¹⁸F]FCH may be collected in vial 250. Valve 160 controls fluid flow to and from QMA 170. Three-way valves 190 and 220 control the flow into and/or out of the silica column 200, waste 230, weak base ion exchange resin 240, and vial 250.

In some embodiments, the methods and processes may be automated by inserting three-way valves 190 and 220 in the illustrated Siemens-CTI Chemical Process Control Unit (CPCU) (CTI, Inc., Knoxville, TN). As is known in the art, a CPCU has three components: chemical process control units, control chasses, operating systems and control software designed to be programmable by implementers to indicate various processes. . As is known in the art, CPCU includes IBM compatible PC systems and Standard (STD) bus subsystems. In thatWindowsOperating System knowsThe Software Package runs the computer thereon (see, e.g., [ 2]¹⁸F]Chemical Process control Unit (chf. sa10.0796.0500, CTI, inc., 1996); and Padget et al, 1989, "Computer-controlled adoptive synthesis: "A chemistry process unit for the automated production of radiopharmaceuticals," int.J.Rad.Appl.Instrum [ A chemistry Process unit for the automated production of radiopharmaceuticals].40(5)：433-45)。

As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Some embodiments are provided by way of illustration or example, and not by way of limitation. It is understood, therefore, that these examples are in no way intended to limit the true scope of the invention. All references cited herein include those patents and/or patent applications in section 1 above, and cross-references to related applications are hereby incorporated by reference.

Specifically, the present invention relates to the following aspects:

1. a method of alkylating a target compound comprising:

a) synthesizing an alkylating agent having the formula:

X-(CR¹R²)_aCR³R⁴-LG

wherein the content of the first and second substances,

a is 0, 1, 2 or 3,

R¹、R²、R³and R⁴Independently H, X or a hydrocarbyl group,

x is halogen or a label, provided that at least one X is halogen,

LG is a leaving group; and

b) directly reacting the alkylating agent with a target compound comprising an alkylating reactive group under conditions suitable for alkylating the target compound.

2. The process of item 1, wherein the alkylating agent has the general formula

X-CH₂-CH₂-LG

Wherein the content of the first and second substances,

x is [ 2]¹⁸F](ii) a And

LG is a sulfonate.

3. The process of item 1, wherein the alkylating agent has the general formula

X-CH₂-LG，

Wherein the content of the first and second substances,

x is [ 2]¹⁸F](ii) a And

LG is a sulfonate.

4. The process of item 1, wherein the alkylating agent has the general formula

CH₃-CH₂(X)-CH₂-LG，

Wherein the content of the first and second substances,

x is [ 2]¹⁸F](ii) a And

LG is a sulfonate.

5. The process of item 1, wherein the alkylating agent has the general formula

X-CH₂-CH₂-CH₂-LG，

Wherein the content of the first and second substances,

x is [ 2]¹⁸F](ii) a And

LG is a sulfonate.

6. The method of item 1, wherein the precursor of the alkylating agent has the general formula LG- (CR)¹R²)_aCR³R⁴-LG。

7. The method of item 1, wherein the precursor has the general formula LG- (CH)₂)₂-LG。

8. The method of item 1, wherein a is 0, R³And R⁴Is a hydrocarbyl group.

9. The method of item 2, 3, 4, 5, 6, 7, or 8, wherein LG is selected from tosylate, mesylate, or triflate.

10. The method of clause 1, wherein the alkylation reactive group comprises an alkyl group, a substituted alkyl group, an alcohol, a carboxylic acid, a saturated cycloalkyl group, an unsaturated cycloalkyl group, an aryl group, a substituted aryl group, a saturated heterocyclic ring, an unsaturated heterocyclic ring, a sulfhydryl group, an amine, N, O, S, or C.

11. The method of item 1, wherein the compound of interest is selected from the group consisting of morphine, heroin, pethidine, tamoxifen, codeine, nicotine, thioproperazine, diazepam, caffeine, flunitrazepam, hexamethonium, methionine, quinuclidinyl diphenylglycolate, MQNB, neostigmine, MPP, NMS, tyrosine, spiperone, and spiroperidol.

12. The method of item 11, wherein the alkylated target compound is [ 2]¹⁸F]fluoromethyl-MQNB [ alpha ], [ alpha¹⁸F]N-fluoromethyl-MPP [ alpha ], [ beta¹⁸F]FNMS、[¹⁸F]Fluoroethyl spiroperone, [ 2]¹⁸F]Fluoromethyl-neostigmine, [ alpha ], [¹⁸F]Fluoromethyl-tyrosine.

13. The method of item 1, wherein the target compound is selected from the group consisting of glucose, lactic acid, cyclohexarbital, thymidine, iodoantipyrine, antipyrine and coenzyme Q.

14. The method of item 1, wherein the target compound is dimethylethanolamine.

15. The method of item 14, wherein the target compound of alkylation is [ alpha ], [ beta ], [¹⁸F]N, N-dimethyl-N-fluoromethylethanolamine.

16. The method of item 14, wherein the target compound of alkylation is [ alpha ], [ beta ], [¹⁸F]N, N-dimethyl-N-fluoroethylethanolamine.

17. Synthesis of¹⁸A method of F-labeling a target compound comprising contacting an alkylating agent having the general formula with a target compound comprising an alkylating group under conditions suitable for alkylating the target compound,

a) synthesis of alkylating agents having the general formula

X-(CR¹R²)_aCR³R⁴-LG

Wherein the content of the first and second substances,

a is 0, 1, 2 or 3,

R¹、R²、R³and R⁴Independently H, X or a hydrocarbyl group,

x is halogen or a label, provided that at least one X is halogen,

LG is a leaving group; and

wherein the target compound of alkylation is different from [ 2]¹⁸F]FCH、[¹⁸F]FECh or [ 2]¹⁸F]Fluorodeoxyglucose.

18. The method of item 17, wherein LG is a sulfonate.

19. The method of item 18, wherein the sulfonate is selected from the group consisting of tosylate, mesylate, and triflate.

20. The method of item 17, wherein the compound of interest is selected from the group consisting of morphine, heroin, pethidine, tamoxifen, codeine, nicotine, thioproperazine, diazepam, caffeine, flunitrazepam, hexamethonium, methionine, quinuclidinyl diphenylglycolate, MQNB, neostigmine, MPP, NMS, tyrosine, spiperone, and spiroperidol.

21. The method of item 20, wherein the alkylated target compound is [ 2]¹⁸F]fluoromethyl-MQNB [ alpha ], [ alpha¹⁸F]N-fluoromethyl-MPP [ alpha ], [ beta¹⁸F]FNMS、[¹⁸F]Fluoroethyl spiroperone, [ 2]¹⁸F]Fluoromethyl-neostigmine, [ alpha ], [¹⁸F]Fluoromethyl-tyrosine.

22. The method of clause 17, wherein the target compound is selected from the group consisting of glucose, lactic acid, cyclohexarbital, thymidine, iodoantipyrine, antipyrine, and coenzyme Q.

23. A process for alkylating dimethylethanolamine comprising:

a) synthesis of an alkylating agent having the formula

¹⁸F-(CH₂)_a-LG

Wherein the content of the first and second substances,

a is 1 or 2, and a is,

LG is a leaving group; and

b) directly reacting said alkylating agent and dimethylethanolamine under conditions suitable to alkylate said dimethylethanolamine.

24. The method of item 23, wherein the alkylating agent has the general formula

¹⁸F-CH₂CH₂-LG

Wherein

LG is a sulfonate.

25. The method of item 23, wherein the alkylating agent has the general formula

¹⁸F-CH₂-LG，

Wherein

LG is a sulfonate.

26. The method of item 23, wherein the precursor of the alkylating agent has the general formula LG- (CH)₂)_a-LG。

27. The method of clauses 23, 24, 25, or 26, wherein LG is selected from tosylate, mesylate, or triflate.

28. The method of item 23, wherein the fluoroalkylated choline is [ alpha ], [ beta ], [¹⁸F]N, N-dimethyl-N-fluoromethylethanolamine.

29. The method of item 23, wherein the fluoroalkylated choline is [ alpha ], [ beta ], [¹⁸F]N, N-dimethyl-N-fluoroethylethanolamine.

30. Computer readable memory for instructing a computer to function in a particular manner, comprising:

a) instructions that direct a computer to execute instructions that control a synthesis module to synthesize an alkylating agent having the general formula:

X-(CR¹R²)_aCR³R⁴-LG

wherein the content of the first and second substances,

a is an integer of 0 to 2,

R¹、R²、R³and R⁴Independently of each other is H or X,

x is halogen or a label, provided that at least one X is halogen,

LG is a leaving group, and

b) instructions that direct the synthesis module to directly react the alkylating agent with a target compound comprising an alkylation reactive group in a reaction vessel under conditions suitable for alkylating the target compound.

31. The computer readable memory of item 30, further comprising:

c) instructions that are executable by the computer to control the synthesis module to bind the alkylated target compound to a first solid support, wherein the reaction vessel and the first solid support are in fluid communication.

32. The computer readable memory of item 31, further comprising:

d) instructions that direct the computer to execute instructions that control the synthesis module to elute the alkylated target compound from the first solid support.

33. The computer readable memory of item 32, further comprising:

e) instructions that are executable by the computer to control the synthesis module to bind the eluted alkylated target compound to a second solid support, wherein the first and second solid supports are in fluid communication, and elute the alkylated target compound from the second solid support.

34.¹⁸A target compound labeled with F, prepared according to the method of item 1 or 17.

35. A composition comprising a compound selected from the group consisting of¹⁸F]Hydrocarbyl morphine, [ 2]¹⁸F]Hydrocarbon heroin [ alpha ], [ alpha¹⁸F]Hydrocarbyl pethidine, [ alpha ], [¹⁸F]Hydrocarbyl tamoxifen [ alpha ], [ beta¹⁸F]Hydrocarbyl codeine [ sic ], [ alpha ]¹⁸F]Hydrocarbyl nicotine [ alpha ], [ alpha¹⁸F]Alkyl sulfur promazine, the term "Alkylthiotetrazine¹⁸F]Hydrocarbyl diazepam [ alpha ], [ beta ], [ alpha ], [ beta ]¹⁸F]Hydrocarbyl caffeine [ sic ], [ alpha ]¹⁸F]Hydrocarbyl flunitrazepam, [ 2]¹⁸F]Alkyl hexamethonium [ sic ], [ solution of ] A¹⁸F]Hydrocarbyl methionine [ alpha ], [ alpha ]¹⁸F]Alkyl diphenylhydroxyacetic acid quinuclidine ester¹⁸F]MQNB、[¹⁸F]Hydrocarbyl neostigmine, [ alpha ], [¹⁸F]alkyl-MPP [ alpha ], [ beta¹⁸F]hydrocarbyl-NMS [ sic ], [¹⁸F]Hydrocarbyl tyrosine, [ 2]¹⁸F]Hydrocarbyl spiroperone and [ 2]¹⁸F]A hydrocarbyl spirocyclic piperidone.

7. Examples of the embodiments

Reagents and solvents were obtained from Aldrich Chemical Co. and Fisher Scientific and were not further purified in use unless otherwise indicated. Melting points were measured on an Electrothermal 9100(Electrothermal engineering Ltd., Southend-on-Sea, UK) and were not corrected. Measurements of NMR spectra were performed on a Varian 400MHz instrument (Varian Inc. Palo Alto, Calif.). Column chromatography was performed using 230Mesh Silica Gel (Catalog No.4791010, Whatman inc., Clifton, NJ). Thin Layer Chromatography (TLC) was performed on Silica 60F254 assay plates (e.merck, Catalog No.4410222, Whatman inc., Clifton, NJ) equipped with a Bioscan 200 imaging scanner (Bioscan, inc., Washington, DC). High Pressure Liquid Chromatography (HPLC) was performed on a Waters 600System (Waters Corporation, Milford, MA) with a UV detector (see, e.g., fig. 15 and 17) (Catalog No. wat080690, Waters Corporation, Milford, MA) and a radioactivity detector (see, e.g., fig. 14 and 16) (Catalog No. fc3200, Bioscan, inc., Washington, DC) connected in series, eluting at 1mL/min with a 20% acetonitrile/water solution containing 0.25mol/L sodium dihydrogen phosphate using a Partisil SCX column (250 x 4.6mm, Catalog No.8173, Alltech Associates, derfield, IL). Gas Chromatography (GC) analysis was performed on a Hewlett-Packard 6890 GC (Hewlett-Packard Company, Palo Alto, Calif.) equipped with a CAM column (30 mm. times.0.25 μm, Catalog No.1122132, J & W Scientific, Agilent Technologies, Inc., Palo Alto, Calif.).

Example 1: xylene sulfonyl methane:

diethylsulfonylmethane was prepared by mixing diiodomethane (1.2g, 4.5mmol) and twice as much of the p-toluenesulfonate silver salt (2.8g, 11mmol) in anhydrous acetonitrile (20 mL). The resulting mixture was refluxed for 16 h. Purification of the xylenesulfonylmethane using 230Mesh Silica Gel Column (Catalog No.4791010, Whatman, inc., Clifton, NJ) (30-40% ethyl acetate-n-hexane) gave the product as white crystals (1g, 63%): m.p.117 deg.C (lit.m.p.116-117 deg.C);¹H-NMR(CDCL₃，400MHz)δ2.45(s，6H，CH₃)，5.10(s，2H，OCH₂O)，7.25(d，J＝8Hz，2H)，7.60(d，J＝8Hz，2H)。

example 2: n, N-dimethyl-N-fluoromethylethanolamine (fluorocholine (FCH)):

a sealed tube containing 10mL of anhydrous Tetrahydrofuran (THF) and 2mL (20mmol) of N, N-dimethylethanolamine was cooled at-78 ℃. Chlorofluoromethane (5.7g, 70mmol, Catalog No.593-70-4, Synquest Labs, Alachua, FL) was bubbled through the cooled solution for 15 minutes. The mixture was allowed to slowly warm to room temperature, held overnight, filtered, and the white solid formed was washed three times with cold THF (-5 ℃) and dried under vacuum. FCH was isolated as a wet white solid (0.5g, 15%):¹H-NMR(D₂O，400MHz)δ3.24(s，3H，CH₃)，3.25(s，3H，CH₃)，3.60-3.63(m，2H，CH₂)，4.05-4.08(m，2H，CH₂OH)，5.43(bd，J＝45Hz，2H，CH₂F)。

example 3: [ ¹⁸ F]Fluoromethane tosylate and [ alpha ], [ alpha ¹⁸ F]Fluoroethane tosylate:

block et al (1987) at J.Label company.radiopharm.24: 1029 ℃ 1042 System on the search for the [1, 1-1, 2-and 1, 3-disubstituted alkanes ] without the addition of a carrier¹⁸F]And (4) oxidizing. We have found that in xylene sulfonyl methane and 1, 2-xylene sulfonyl2 of ethylene¹⁸F]The results of Block et al were reproduced in the fluorination. Both reactions were heated at 80 ℃ for 5 min and the products were analyzed by TLC. TLC was performed on Silica 60F254 assay plates (Catalog No.4410222, Whatman inc., Clifton, NJ) in 30% ethyl acetate/n-hexane. In these studies, the term¹⁹F]Fluoride provides a UV detectable moiety for quality control analysis.

1, 2-ditosylethane¹⁸F]Fluorination to give the product in 80% yield¹⁸F]Fluoroethane tosylate (fig. 1). No detection of [ 2]¹⁸F][¹⁹F]Difluoroethane. Therefore, since¹⁸F]Fluorine is in the beta position and no electron withdrawing effect is observed. A yield of 80% was obtained without stirring. Can be prepared at a lower temperature¹⁸F]Fluoroethane tosylate, but one of ordinary skill in the art will recognize that yield decreases with decreasing temperature.

Of xylene sulfonyl methane under various conditions¹⁸F]Fluorination to obtain two labeled products¹⁸F]Difluoromethane and [ 2]¹⁸F][¹⁹F]Difluoromethane tosylate. At 80 ℃, only one product is formed, and the synthesis [ 2]¹⁸F]The unused byproduct of fluorocholine [ 2]¹⁸F][¹⁹F]Difluoromethane. In addition, some free detectable¹⁸F]Fluoride is not common, especially when the reaction is not complete. As expected by the Block et al study, 2¹⁸F]The yield of fluoromethane tosylate was 1% (FIG. 2). [¹⁸F]The low yield of fluoromethane tosylate may be due to the electron withdrawing effect of fluorine at the alpha position at SN₂In the reaction¹⁸F]Fluoromethane tosylate is more reactive than ditosylmethane. Since a very low concentration of [ 2] is detected after the reaction¹⁸F]Fluoride, apparently [ 2]¹⁸F]Fluoromethane tosylate and [ 2] present in the reaction vessel¹⁹F]The carrier may be subjected to a second fluorination to give a solution of¹⁸F][¹⁹F]Difluoromethane, a very volatile compound (boiling point-52 deg.C))。

We examined the effect of temperature on the fluorination of xylenesulfonylmethane. When the reaction temperature is lowered to about 45 to 50 ℃¹⁸F][¹⁹F]The production of difluoromethane is decreased, and we obtained a sufficient amount of [ 2], [¹⁸F]Synthesis of fluoromethane tosylate [ alpha ], [ alpha¹⁸F]FCH (fig. 2). At a temperature of less than about 40 ℃ for up to 20 minutes, only a small amount of the [ alpha ], [ beta¹⁸F][¹⁹F]Difluoromethane; however, this reaction rate is not suitable for radiosynthesis using some short-lived isotopes. At a temperature greater than about 50 ℃, the predominant reaction product is [ 2]¹⁸F][¹⁹F]Difluoromethane.

The fluorination step is also sensitive to agitation. In the absence of agitation at a temperature in the range of about 40 ℃ to about 50 ℃, the reaction gives a small amount of¹⁸F][¹⁹F]Difluoromethane but also a small amount of¹⁸F]Fluoromethane tosylate (. about.10% yield). Is carried out while stirring at a temperature in the range of about 40 ℃ to about 50 ℃¹⁸F]When fluorinated, a sufficient amount of¹⁸F]Fluoromethane tosylate (yield 30%) to synthesize [, ]¹⁸F]FCH。

Example 4: [ ¹⁸ F]N, N-dimethyl-N-fluoromethylethanolamine ([ 2], ] ¹⁸ F]FCH)：

Prepared in two steps¹⁸F]FCH: use 2¹⁸F]Fluoride Fluoroxylenesulfonylmethane followed by [ 2] using a modified Siemens-CTI chemical Process control Unit (CPCU, Catalog No.3601037, CTI, Knoxville, TN) (FIG. 10)¹⁸F]Alkylation of fluoromethane tosylate with dimethylethanolamine. Purification [ 2], (Silica Sep-Pak column (Catalog No. WAT023537, Waters Corporation, Milford, MA)¹⁸F]FCH. The column was washed with ethanol and water to remove all impurities, and eluted with 2% acetic acid¹⁸F]FCH. Using AG 4-X4 weakly basic ion exchange resinColumn (143. sub.3341, Bio-Rad laboratories, Inc., Hercules, Calif.) removes acetic acid (see AG 4 Induction Manual LIT207 Rev B, Bio-Rad). Two three-way slide valves, such as teflon slide valves, were added to the CPCU for purification (fig. 10). The software was modified to lower the temperature and allow stirring during the labeling step.

To prepare [ 2]¹⁸F]Fluoride, rich in¹⁸O-water was irradiated with 11MeV protons (33. mu.A, 60 min). After proton bombardment, the target was pressed using Ar gas pressure¹⁸F-HF/¹⁸The solution of O-water was delivered to an anion exchange QMA cartridge (cartridge) (carbonate form) (Catalog No. wat054725, waters corporation, Milford, MA). The [ 2] without adding a carrier¹⁸F]Fluoride capture on QMA cartridges and recovery¹⁸O-water. 1mL of potassium carbonate (K) containing 2.2mM₂CO₃) And 40mM2.2.2(4, 7, 13, 16, 21, 24-hexaoxa-1, 10-diazabicyclo [ 8.8.8)]Hexacosane, C₁₈H₃₆N₂O₆Aqueous acetonitrile solution (19: 1) of Catalog No.291110, Sigma-Aldrich Corp, St. Louis, Mo¹⁸F]Fluoride elutes from the cartridge to the reaction vessel. The mixture was azeotropically dried at 110 ℃ under Ar gas flow for 4 minutes and allowed to cool for about 60 seconds. Diethylsulfonylmethane (10mg, 0.03mM) in 1mL of anhydrous acetonitrile was added to the dried residue. The mixture was heated at 45-50 ℃ for 5 minutes while intermittently bubbling Ar gas (10 seconds per 20 seconds). Thereafter, 1mL of acetonitrile containing 0.2mL of N, N-dimethylethanolamine was added to the reaction vessel, and heated at 100 ℃ for 10 minutes while intermittently bubbling Ar gas (15 seconds per 30 seconds) (FIG. 3). Charging the entire mixture into SiO₂Sep-Pak cartridges (Catalog No. WAT023537, Waters Corporation, Milford, MA) and washed with ethanol (3X 5mL) and water (2X 5 mL). The effluent was introduced into the final product vial using two three-way valves. With 5mL of 2% acetic acid followed by 5mL of H₂O from SiO₂Released from the Sep-Pak cartridge¹⁸F]FCH. Passing through AG 4-X4 weakly basic ion exchange resinColumn (Catalog No.143-3341, Bio-Rad laboratories, Inc., Hercules, Calif.) (see AG 4 Instruction Manual LIT207 RevB, Bio-Rad) acetic acid was stripped from the eluate and the solution passed through a 0.2 μm membrane filter (Catalog No. SLGS V255F, Millipore Corp, Billerica, Mass.) into a sterile vial containing 0.5ml of 23.4% concentrated sodium chloride. Alternatively, SiO was replaced by a previously activated (10mL of 1N HCl) Accell cartridge (CatalogNo. WAT023531, Waters Corp, Millford, Mass.)₂Sep-Pak cartridge, and eluted with 10mL of saline¹⁸F]FCH。

The result of the alkylation reaction indicates that in acetonitrile¹⁸F]The fluoromethane tosylate and N, N-dimethylethanolamine were almost quantitatively reacted to give a yield of > 90%¹⁸F]FCH。[¹⁸F]The fluoromethane tosylate is synthesized as the [ 2]¹⁸F]The total yield of FCH is 20%, which may be due to the production of [ 2], [ 2] during the fluorination step¹⁸F][¹⁹F]Difluoromethane. The total synthesis time is less than about 40 minutes. (see FIGS. 16 and 17).

Example 5: synthesis of N, N-dimethyl-N-fluoroethylethanolamine (fluoroethylcholine (FECh)):

1-bromo-2-fluoroethane (1g, 8mmol) was dissolved in N, N-dimethylethanolamine (0.7g, 8mmol) and heated at 100 ℃ for 10 minutes. The resulting mixture was dissolved in 10% ethanol/ethyl acetate. After removal of the solvent, FECh was obtained as a white solid (1.7g, 100%).¹H-NMR spectra and Hara et al 2002, J.AM Med.43 (2): the spectrum reported in 187-.

Example 6: [ ¹⁸ F]N, N-dimethyl-N-fluoroethylethanolamine ([ 2], ] ¹⁸ F]FECh) synthesis:

anhydrous [ alpha ], [ beta ] is obtained as described in example 3¹⁸F]Fluoroideneand reacted with 1, 2-bis-xylenesulfonylethane (10mg, 0.03mmol) in 1mL of anhydrous acetonitrile. At 80 deg.CThe mixture was heated for 5 minutes while intermittently bubbling Ar gas (10 seconds per 20 seconds). Thereafter, 0.3mL of pure N, N-dimethylethanolamine was added to the reaction vessel, and heated at 100 ℃ for 10 minutes while intermittently bubbling Ar gas (15 seconds per 30 seconds). The reaction mixture was purified from the reaction mixture using a Silica GelSep-Pak cartridge as described in example 3¹⁸F]FECh。

Alkylation of fluoroethane tosylate in pure N, N-dimethylethanolamine gave only 50% yield. [¹⁸F]The total yield of FECh is also 50%, [ 2]¹⁸F]The FCH yield (20%) is high. This is probably because very little [ 2] is produced during the fluorination step¹⁸F][¹⁹F]By-production of difluoroethane or no production of¹⁸F][¹⁹F]Difluoroethane by-product. As described above¹⁸F]FCH describes that the total synthesis time is less than about 40 minutes (see fig. 14 and 15).

Example 7: [ ¹⁸ F]FCH and [ 2] ¹⁸ F]Purification of FECh

[¹⁸F]FCH and [ 2]¹⁸F]FECh are quaternized hydrocarbyl amines that can be adsorbed strongly on Silica GelSep-Pak cartridges (Mulholland et al, (1999) J. Label components radiopharmam.42: suppl.1s459-s 461). [¹⁸F]FCH and [ 2]¹⁸F]The FECh adheres strongly to the cartridge, which allows most of the impurities to be quickly removed as waste during ethanol elution. Eluted with 2% acetic acid¹⁸F]FCH and [ 2]¹⁸F]FECh, and neutralized using a weakly basic ion exchange resin (AG 4-X4, Catalog No.140-4341, Bio-Rad Laboraotires, Inc., Hercules, Calif.).

[¹⁸F]FCH and [ 2]¹⁸F]Emission chemical purity of FECh was determined by HPLC (HPLC 600System, Waters Corporation, Milford, Mass.) and TLC (Silica 60F254 Analytical plants, Whatman Inc., Clifton, NJ). [¹⁸F]FCH and [ 2]¹⁸F]FECh from HP at 6.8 min and 7.5 min, respectivelyEluting in an LC column. HPLC was performed on a Waters 600System (Waters Corporation, Milford, Mass.) with a UV detector (Catalog No. WAT080690, Waters Corporation, Milford, Mass.) and a radioactivity detector (Catalog No. FC3200, Bioscan, Inc., Washington, DC) connected in series using a Partisil SCX column (250X 4.6mm, CatalogNo.8173, Alltech Associates, Deerfield, IL) eluting with a 20% acetonitrile/water solution containing 0.25mol/L sodium dihydrogen phosphate at 1 mL/min. The final solution was analyzed for residual amounts of N, N-dimethylethanolamine by GC using a CAM column designed for amines (Catalog No.1122132, Agilent Technologies, Inc., Palo Alto, Calif.). The only chemical impurities detected by HPLC and GC were N, N-dimethylethanolamine (< 0.5 mg/lot) and ethanol (< 5 mg/lot).

To reduce the contamination of N, N-dimethylethanolamine, the Silica Gel Sep-Pak cartridge was replaced by an Accell cartridge (Hara et al (1997) J.Nucl.Med.38 (6): 842. sup.847). With HCl and H₂O preconditions the Accell cartridge and the final product is eluted with brine solution. In [ 2]¹⁸F]N, N-dimethylethanolamine < 0.1mg per batch was found in the preparation of FCH. Cannot be sufficiently purified using Accell cartridge¹⁸F]FECh, i.e., about half of the value found in the waste fraction¹⁸F]FECh。

The final purified product in the culture physiological saline¹⁸F]FCH and [ 2]¹⁸F]FECh to grow microorganisms in thioglycolate enriched at 37 ℃ and trypticase casein soy broth medium at 23 ℃. No growth was observed within two weeks. Standard methods using LAL (Charles River Labs Endosafe, Charleston, SC) indicated that the final purified solution was pyrogen-free. Positive control standards are often included in the test. [¹⁸F]FCH is currently used in FDA approved pre-clinical IND safety trials in humans. To date, after about 50 experiments, no test was observed¹⁸F]Fluorocholine has adverse effects.

TLC plates were pretreated in 2% acetic acid and air dried before use. The plate was eluted in 2% acetic acid. R_f[¹⁸F]FCH and [ 2]¹⁸F]FECh of 0.4, confirmedThe HPLC results are shown.

Example 8: other alkylating agents, alkylation processes and purification processes

Alkylating agents

Used for preparing¹⁸F]FCH and [ 2]¹⁸F]SN of FECh₂Alternatives to tosylate leaving groups for nucleophilic substitution reactions include, but are not limited to, halides and other sulfonates, for example, methane sulfonates (e.g., mesylate) and trifluoromethane sulfonates (e.g., triflate). However, as the volatility of the alkylating agent increases, the retention of the alkylating agent in solution decreases. Thus, volatility may be reduced by increasing the number of carbon atoms so that the alkylating agent may remain in the solvent.

Use 2¹⁸F]The fluoride labeled 1, 2-bis (trifluoromethanesulfonyl) ethane was obtained in a yield of > 80%¹⁸F]Fluoroethane triflate. The yield is greater than 2¹⁸F]The yield of fluoroethane tosylate was because triflate was the most reactive of the three sulfonates (mesylate, tosylate, triflate). Furthermore, higher yields were obtained in the alkylation step with N, N-dimethylethanolamine using triflate as leaving group. Use of 2-¹⁸F]Alkylation of fluoroethane tosylate and N, N-dimethylethanolamine gave 2 [, [ 2] in 50% yield¹⁸F]FECh. In contrast, 2 [ - ]¹⁸F]Reacting fluoroethane triflate with N, N-dimethylethanolamine to obtain a quantitative product¹⁸F]FECh. Thus, the [ 2], [1, 2-bis (trifluoromethanesulfonyl) ethane, which is obtained using it as an unlabeled precursor¹⁸F]The overall yield of FECh was > 70%.

As a term for us¹⁸F]As a result of investigation of the fluoride-labeled xylenesulfonylmethane, the second fluorination due to the alpha position¹⁸F]The electron withdrawing effect of fluorine is activated, resulting in the synthesis of difluoromethane. Therefore, it would be advantageous to slow or prevent the second active fluorination step. Polymer resins containing covalently bonded quaternary ammonium salts(see, e.g., J.Labelled.Cpd.radiopharmaceuticals 26: 378-¹⁸O]H₂O [ solution of [ 2] without addition of a carrier¹⁸F]Fluoride and mark xylenesulfonylmethane. Polymer resin treatments may not be as efficient as solution reactions, but they do slow or prevent¹⁸F][¹⁹F]Formation of difluoromethane.

Alkylation:

because the sulfonate is more active than the halide ester¹⁸F]Fluoromethane tosylate and [ alpha ], [ alpha¹⁸F]Fluoroethane tosylate is suitably subjected to on-column alkylation (on-column alkylation) with N, N-dimethylethanolamine. The sulfonate was trapped on Sep-Pak C18 cartridge loaded with dimethylethanolamine. Thus, of N, N-dimethylethanolamine¹⁸F]Fluoromethylation is improved (streamlined) and the alkylation step is faster.

And (3) purification:

we have described carrying out the process using a Silica Gel or Accell cartridge¹⁸F]And (4) FCH purification. However, the term¹⁸F]FECh was well purified on Silica Gel but not on Accell cartridges. This may be due to an alkylation step which requires the use of undiluted (pure) N, N-dimethylethanolamine. For [ 2]¹⁸F]FCH, alkylation takes place in the solvent acetonitrile. Thus, the [ alpha ]¹⁸F]Prior to the FECh reaction mixture, 1mL of acetonitrile was added, which allowed the reaction to proceed¹⁸F]The FECh remains on the Accell cartridge, facilitating purification.

Example 9: others [ 2]¹⁸F]-labelled target compound

The disclosed method can be used to mark halogens with longer lifetimes (e.g., using a label with a longer lifetime¹⁸F) The equivalent compound of (a) instead of the label is [1 ]¹¹C]A target compound of a hydrocarbyl iodide. Thus, the [ 2]¹⁸F]Fluoroalkanesulfonates can be used for alkylation of a variety of N, O, S and C. Can be obtained by the method disclosed by the inventionCarry out¹⁸F]Examples of fluoroalkylated quaternary amines include MQNB (3-quinuclidine diphenylglycolate derivative), a muscarinic ligand (figure 5); the acetylcholinesterase inhibitor neostigmine (fig. 6) and its metabolite TMA-phenol; neurotoxin N-methyl-4-phenyl-pyridinium labeled using this method. Can be prepared according to the method disclosed in the present invention¹⁸F]The secondary amine subjected to fluoromethylation comprises [, [ 2]¹⁸F]N-Methylspirone (¹⁸F]NMS) and 3- (2' - [ solution ]¹⁸F]Fluoroethyl spiroperone (¹⁸F]FESP) (fig. 7). The O-alkylation reaction was used to methylate the amino acid tyrosine in the presence of dimethyl sulfoxide (DMSO) (FIG. 8). The carboxylic acid may be alkylated as described in figure 9. S-alkylation the target compound of FIG. 9 was fluoromethylated.

Claims (13)

1. Synthesis of¹⁸A method of labeling a target compound comprising

a) Synthesis of alkylating agents having the general formula

X-(CR¹R²)_aCR³R⁴-LG

Wherein the content of the first and second substances,

a is 0, 1, 2 or 3,

R¹、R²、R³and R⁴Independently is H or a hydrocarbon group,

x is¹⁸F，

LG is a leaving group; and wherein the alkylating agent is synthesized by halogenating a precursor thereof, wherein the temperature of the halogenation reaction is from 45 ℃ to 50 ℃, and

b) contacting the alkylating agent with a target compound comprising an alkylation reactive group under conditions suitable for alkylating the target compound, wherein the alkylated target compound is different from¹⁸F]FCH、[¹⁸F]FECh or [ 2]¹⁸F]Fluorodeoxyglucose, and wherein said contacting is effected without purification of said alkylating agent after said synthesizing.

2. The method of claim 1, wherein LG is a sulfonate.

3. The method of claim 2, wherein the sulfonate is selected from the group consisting of tosylate, mesylate and triflate.

4. The method of claim 1, wherein the target compound is selected from the group consisting of morphine, heroin, pethidine, tamoxifen, codeine, nicotine, thioproperazine, diazepam, caffeine, flunitrazepam, hexamethonium, methionine, 3-quinuclidinyl diphenylglycolate, neostigmine, 4-phenyl-pyridinium, spiperone, tyrosine, and spiroperidol.

5. The method of claim 4, wherein the alkylated target compound is [ alpha ], [ beta ]¹⁸F]Fluoromethyl-diphenylglycolic acid 3-quinuclidine ester, [ 2]¹⁸F]N-fluoromethyl-4-phenyl-pyridinium salt [ sic ], [ solution ]¹⁸F]N-fluoromethylspirone [ alpha ], [ beta ], [ alpha ], [ beta ]¹⁸F]Fluoroethyl spiroperone, [ 2]¹⁸F]Fluoromethyl-neostigmine.

6. The method of claim 1, wherein the target compound is selected from the group consisting of glucose, lactic acid, cyclohexarbital, thymidine, iodoantipyrine, antipyrine and coenzyme Q.

7. A process for alkylating dimethylethanolamine comprising:

a) synthesis of an alkylating agent having the formula

¹⁸F-(CH₂)_a-LG

Wherein the content of the first and second substances,

a is 1 or 2, and a is,

LG is a leaving group, wherein the alkylating agent is synthesized by halogenating a precursor thereof, wherein the temperature of the halogenation reaction is from 45 ℃ to 50 ℃; and

b) directly reacting said alkylating agent and dimethylethanolamine under conditions suitable for alkylating said dimethylethanolamine, wherein said contacting is accomplished without purifying said alkylating agent after performing said synthesis.

8. The process of claim 7 wherein said alkylating agent has the formula

¹⁸F-CH₂CH₂-LG

Wherein

LG is a sulfonate.

9. The process of claim 7 wherein said alkylating agent has the formula

¹⁸F-CH₂-LG，

Wherein

LG is a sulfonate.

10. The process of claim 7, wherein the precursor of the alkylating agent has the general formula LG- (CH)₂)_a-LG。

11. The method of claim 7, 8, 9, or 10, wherein LG is selected from tosylate, mesylate, or triflate.

12. The process of claim 7, wherein the fluoroalkylated choline is [ [ solution ], [ solution ] ]¹⁸F]N, N-dimethyl-N-fluoromethylAnd (3) monoethanolamine.

13. The process of claim 7, wherein the fluoroalkylated choline is [ [ solution ], [ solution ] ]¹⁸F]N, N-dimethyl-N-fluoroethylethanolamine.