Improved synthesis of radiolabeled prostate-specific membrane antigen (PSMA) inhibitor [18f]dcfpyl

Abstract

To provide a method of synthesizing 2-(3{1-carboxy-5-[(6-[18F]fluoro-pyridine-3-carbonyl)-amino]-pentyl}-ureido)-pentanedioic acid ([18F]DCFPyL), which is a radiotracer for prostate-specific membrane antigen (PSMA) PET imaging for prostate cancer.SOLUTION: A synthesis method comprises the following steps: (i) radiofluorinating a DCFPyL precursor comprising ester moiety-protecting groups to form a radiofluorinated DCPFPyL precursor; (ii) deprotecting the ester moiety-protecting groups of the radiofluorinated DCPFPyL precursor of the step (i) with phosphoric acid to form [18F]DCFPyL in a reaction mixture; and (iii) purifying the [18F]DCFPyL from the reaction mixture of the step (ii) to provide [18F]DCFPyL.SELECTED DRAWING: Figure 5

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 62 / 348,391, filed June 10, 2016. No. 6,299,499, filed on Oct. 1, 2003, which is incorporated herein by reference in its entirety.

[0002] Federally funded research and development This invention was made in part thanks to grants CA134675 and CA183031 awarded by the National Institutes of Health (NIH). This invention was made with government support under the Federal Trade Commission. The U.S. Government has certain rights in this invention. [Technical Field]

[0003] The present invention relates to 18 The present invention relates to methods and related compositions for the improved synthesis of [F]DCFPyL. Manufactured by [ 18 Methods and related compositions for using [F]DCFPyL are also provided. [Background technology]

[0004] Prostate cancer has an estimated annual incidence of over 1 million cases and an estimated annual death toll of 307,000. , the most common cancer in men and one of the most prevalent cancers worldwide (Mauer et al., 2016). In the United States alone, over 200,000 new cases are diagnosed each year. In advanced prostate cancer, prostate-specific antigen (PSA) Thanks in part to serodiagnostic testing for expression, appropriate diagnosis and treatment can improve 5-year survival. The rate is nearly 99% (seer.cancer.gov).

[0005] Adequate diagnosis and monitoring of treatment requires more frequent, non-invasive Molecular imaging includes prostate-specific membrane antigen (PSMA) PET imaging for prostate cancer. A number of radioactive tracers have been developed for imaging, including DCFBC (Mease et al. , 2008) and DCFPyL ( Chens et al., 2011 ) [ 11 C] Choline, 18 F]fluorocholine, 6 8 Ga]- and [ 18 Other PSMA-targeting radiopharmaceuticals include [F]-labeled low-molecular-weight PSMA inhibitors. Compared to radioactive tracers (Dietlein et al., 2015), 18 F] DCFPyL (Chen et al., 2011; Szab o et al., 2015) is used to achieve successful treatment, as well as its good distribution and imaging. The demand for this radioactive tracer has increased due to its therapeutic properties. Summary of the Invention [Means for solving the problem]

[0006] Certain embodiments described herein include 2-(3-{1-carboxy-5-[(6-[ 18 F]fluoro-pyridine- 3-carbonyl)-amino]-pentyl}-ureido)-pentanedioic acid ([ 18 F]DCFPyL) In one aspect, the method comprises: (i) forming a radiofluorinated DCPFPyL precursor; (ii) radiofluorination of a DCFPyL precursor containing an ester moiety protecting group; ) in the reaction mixture [ 18 radiofluorination of DCPFPyL in step (i) to form [F]DCFPyL (iii) deprotecting the ester moiety of the precursor with phosphoric acid; 18 F]DCFPyL To do this, from the reaction mixture of step (ii) 18 and purifying [F]DCFPyL.

[0007] In some embodiments, the ester moiety protecting group of the DCFPyL precursor includes benzyl, p-methoxybenzyl, butyl (PMB), tertiary butyl (tert-butyl or t-butyl), methoxymethyl (MOM), meth ethoxymethyl (MEM), methylthiomethyl (MTM), tetrahydropyranyl (THP) , tetrahydrofuranyl (THF), benzyloxymethyl (BOM), trimethylsilyl (TM S), triethylsilyl (TES), t-butyldimethylsilyl (TBDMS) and triphenylmethylsilyl Protecting groups include those selected from the group consisting of methyl (trityl, Tr), methyl (C1), methyl (C2), methyl (C3), methyl (C4), methyl (C5), methyl (I), methyl (I6), methyl (I7), methyl (I8), methyl (I9), methyl (I10), methyl (I11), methyl (I12), methyl (I13), methyl (I14), methyl (I15), methyl (I1 In one embodiment of any one of the methods, the ester moiety protecting group of the DCFPyL precursor includes a tert -butyl groups.

[0008] In one embodiment of any one of the methods described herein, radiofluorination and defluorination are performed. Protection may be carried out in a single reactor.

[0009] The present invention relates to the 18 The synthesis of [F]DCFPyL can be carried out manually or automatically. In some embodiments of any of the methods described herein, hand,[ 18 The synthesis of [F]DCFPyL is carried out using a heating block, syringe pump (e.g., at least 2 syringe pump), multi-port cap, and valved reagent addition vial It is automated by using a radiofluorination module (RFM) including In some embodiments, the RFM may further comprise thermal heating. In another embodiment of any of the described methods, 18 The synthesis of [F]DCFPyL was performed using an ELIXYS autoradiometer. Chemical synthesis apparatus (Sophie Biosciences, Culver City, CA) may be automated using Sofie Biosciences, Inc., Culver City, CA) In some embodiments, the RFM or ELIXYS automated radiochemical synthesizer components contain fluorine. Prior to synthesis, the RFM or ELIXYS automated radiochemical synthesizers, or The reaction area may be cleaned with dilute nitric acid, washed with water, and dried at 80°C overnight.

[0010] Any known DCFPyL precursor can be subjected to a radiofluorination process according to the methods described herein. In one embodiment of any one of the methods provided herein, , wherein the DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy) -1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carbamoyl)-N,N,N- In one embodiment, the DCFPyL precursor is trimethylpyridin-2-aminium. , 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopenta N,N,N-trimethylpyridine-2-yl)ureido)-6-oxohexyl)carbamoyl aminium trifluoromethanesulfonate. In another embodiment, the DCFPy The L precursor is 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-di (2-hexopentanyl)ureido)-6-oxohexyl)carbamoyl)-N,N,N-trimethyl Pyridine-2-aminium trifluoroacetate.

[0011] In certain embodiments of any one of the methods provided herein, the DCFPyL The precursor was N,N,N-trimethyl-5-((2,3,5,6-tetrafluorophenoxy)carbonyl) -pyridin-2-aminium trifluoromethanesulfonate (compound (2) shown in Figure 1) 2-{3-[1-t-butylcarboxylate-(5-aminopentyl)]-uriedo}-di-t-butyl by a method involving coupling with ethyl pentane diate (compound (1) shown in Figure 1) In one embodiment of any one of the methods described herein, the DCFPyL precursor is synthesized. Synthesize according to Figure 1.

[0012] In certain embodiments of any one of the methods provided herein, the DCFPyL precursor is prepared by subjecting the DCFPyL precursor to a process as shown in FIG. Radiofluorination is carried out according to the method described above.

[0013] In one embodiment of any one of the methods provided herein, 18 [F]DCFPyL was subjected to QC. It has characteristics that meet the input specifications (2016).

[0014] In some embodiments of any one of the methods provided herein, the DCFPyL precursor is treated by: (a) [ 18 (b) capturing [F] fluoride ions in said cartridge; Captured 18 To release the [F] fluoride ion, tetrabutylammonium base salts (e.g. For example, a solution of tetrabutylammonium bicarbonate (TBABC) is used to remove the ammonium hydroxide from the cartridge. (c) drying 18 to form a fluoride ion [F] 18 F] fluoride (d) drying the eluate containing the ions; 18 F] fluoride ion, D Adding a solution of CFPyL precursor (compound (3) shown in Figure 5) and radioactively fluorinated.

[0015] In one embodiment of any one of the methods described herein, 18 [F] fluoride ion The capture cartridge is an anion exchange chromatography cartridge (e.g., For example, Chromafix 30-PS-HCO3 SPE cartridges (source). In one embodiment of any one of the methods, the cartridge is 18 F] Before capturing fluoride ions, the filter can be preconditioned by washing with high-purity water. be.

[0016] In some embodiments of any one of the methods described herein, the [ 18 F] Dry the fluoride ions. 18 to generate the fluoride ion [F] In one embodiment, the 18 The eluate (c) containing [F] fluoride ions is heated at a temperature of about 80 °C to about 150 °C. The drying may be carried out at a temperature, for example, about 110° C. 18 F] fluoride ion The eluate of step (c) may be dried under a stream of nitrogen. The process may last from about 50 seconds to about 300 seconds, or more preferably, 150 seconds. In one embodiment, the dried [ 18 [F] fluoride ion in CH3CN may be added.

[0017] In some embodiments of any one of the methods provided herein, the DCFPyL precursor is dried. Seta [ 18 [F] fluoride ion, the combined solution is heated, for example, at about 30°C to In one embodiment, the heating is carried out for about 2 minutes to about 10 minutes. In one embodiment, the heating is performed at about 50°C for about 6 minutes. This can be done by any known method, but in one embodiment, it is done by drying the DCFPyL precursor. Ta [ 18 The mixed solution with [F] fluoride ion was irradiated with microwaves at about 40 W to about 60 W. The heating is carried out by irradiating the substrate with a light source for about 20 seconds to about 200 seconds. CFPyL precursor and dried [ 18 The mixed solution containing [F] fluoride ions was heated at approximately 50 W in a microwave oven. The heating is performed by irradiating for about 30 seconds to about 150 seconds.

[0018] The DCFPyL precursor is [ 18 After reaction with [F] fluoride ions, the esters of the resulting products The protecting group of the aryl moiety is deprotected with phosphoric acid. In some embodiments, the deprotection is carried out at a temperature of about 30° C. to about 55° C. is carried out at a temperature of about 45° C. In some embodiments of any one of the methods described herein, The deprotection is carried out for about 2 to about 10 minutes at the desired temperature.

[0019] In certain aspects of any one of the methods provided herein, the method comprises deprotection with phosphate. After the reaction, the method further comprises adjusting the pH of the reaction mixture to a pH between about 2 and about 2.5. Examples of buffers that can be used to adjust the pH of the reaction mixture include, but are not limited to: Examples of suitable buffers include, but are not limited to, sodium hydroxide and sodium dihydrogen phosphate buffers.

[0020] [ 18 [F]DCFPyL can be purified using any purification and separation method known in the art. In one embodiment of any one of the methods provided herein, 18 Purification of [F]DCFPyL This is done by liquid chromatography. For example, 18 F]DCFPyL at least one C1 Purification may be achieved by liquid chromatography using an 8 column. , [ 18 The solution containing [F]DCFPyL was removed from the first C18 column with methanol and sodium dihydrogen phosphate. Elute with the first elution solution containing thorium. Example of methanol vs. sodium hydrogen phosphate In one embodiment, the sodium dihydrogen phosphate is added in an amount of about 0.01:1. The solution may be prepared at a concentration of 0.1 M (pH 2.1) by any one of the methods provided herein. In this aspect, 18 The solution containing [F]DCFPyL was then passed through a second C18 column and purified with alcohol. The protein is eluted with a second elution solution containing ethanol (e.g., ethanol).

[0021] In certain embodiments of any one of the methods provided herein, the first or second C18 column Eluted from [ 18 The [F]DCFPyL may be further subjected to filtration. Filtration is carried out using a 0.2 μm sterile filter. 18 F]DCFPyL is optionally Alternatively, the solution may be filtered directly into a sterile vial prefilled with sterile saline.

[0022] In one embodiment of any one of the methods provided herein, 18 F]DCFPyL purification process The reaction can be carried out in the presence of sodium ascorbate. 18 F]DCFP A collection reservoir and / or a column used to elute the yL product, e.g., from a C18 column. Sodium ascorbate may be added to the solution.

[0023] In one embodiment, the [ 18 The method for synthesizing [F]DCFPyL is to obtain high amounts of [ 18 F]D In one embodiment, the purified CFPyL 18 The yield of [F]DCFPyL is at least 20 mCi In one embodiment, the purified 18 The yield of [F]DCFPyL is at least 100 mCi. In this embodiment, the purified 18 The yield of [F]DCFPyL is at least 400 mCi.

[0024] Another aspect provided herein relates to a method for radiofluorinating a DCFPyL precursor. (i) the cartridge contains [ 18 (ii) capturing [F] fluoride ions; Captured in the cartridge 18 Tetrabutylammonium chloride is used to release the [F] fluoride ion. (iii) eluting the cartridge with a solution of tetrahydrobenzoate (TBABC); Origin of [ 18 (iv) drying the eluate containing [F] fluoride ions; [ 18 A solution of DCFPyL precursor (e.g., compound (3) shown in Figure 5) is added to the [F] fluoride ion. and (v) heating the mixed solution of step (iv).

[0025] In one embodiment of any one of the methods described herein, 18 [F] Fluoride ion capture The cartridge for this purpose is an anion exchange chromatography cartridge (e.g., Ch Romafix 30-PS-HCO3 SPE cartridges). In one embodiment, the cartridge is 18 [F] Fluoride ion capture Before use, the sample may be preconditioned by rinsing with high purity water.

[0026] In some embodiments of any one of the methods described herein, the method further comprises the step of: 18 F] Fluoride Dry the on. Dry [ 18 [F] fluoride ions, in some embodiments: The above [ 18 The eluate (c) containing [F] fluoride ions is heated at a temperature of about 80°C to about 150°C, e.g. For example, the drying may be performed at about 110°C. 18 (c) containing fluoride ions [F] The eluate may be dried under a stream of nitrogen. In some embodiments, the drying process is carried out for about 50 minutes. The period may last from about 100 seconds to about 300 seconds, or more preferably about 150 seconds. , and further drying the dried [ 18 Adding CH3CN to [F] fluoride ions There is a saying.

[0027] In some embodiments of any one of the methods provided herein, the DCFPyL precursor is dried. Seta [ 18 [F] fluoride ion, the combined solution is heated, for example, at a temperature of from about 30° C. to about 7 0°C. In some embodiments, the heating may be performed for about 2 minutes to about 10 minutes. In one embodiment, the heating is performed at about 50° C. for about 6 minutes. The heating may be performed by any method known in the art. This can be done by any method, but in one embodiment, the DCFPyL precursor and dried [ 1 8 The mixed solution with [F] fluoride ions was heated at a power of about 40 W to about 60 W for about 20 seconds to about 200 In one embodiment, the DCFPyL precursor and the dried Seta [ 18 The mixed solution with [F] fluoride ions was irradiated with microwaves at approximately 50 W for approximately 30 seconds to The heating is carried out by irradiating with microwaves for about 150 seconds.

[0028]

[0023] Produced by any one of the methods described herein 18 Compositions containing F]DCFPyL In one embodiment of any one of the compositions described herein, 18 DCF PyL has an average specific activity of at least about 50 Ci / μmole. In one embodiment of the composition, 18F]DCFPyL is at least about 50 Ci / μmole or less They have an average specific activity of at least about 100 Ci / μmole.

[0029] In one embodiment of any one of the compositions described herein, 18 Radiochemical analysis of [F]DCFPyL The purity ranges from about 95% to about 100%. In some embodiments, the composition comprises acetonitrile at a concentration of about 400 ppm or less. In one embodiment of any one of the compositions described herein, the composition has a concentration of about 3,000 ppm or less. In one embodiment of any one of the compositions described herein, The composition contains methanol at a concentration of about 50 ppm or less.

[0030] In certain embodiments of any one of the compositions described herein, the composition comprises one or more clitorises. and / or a methacrylate copolymer, e.g., containing one or more Kryptofix® compounds. In one embodiment of any one of the compositions described herein, the composition does not contain triethylene glycol. In one embodiment of any one of the compositions described herein, the composition is The product does not contain t-butanol.

[0031] Also provided herein are kits containing any one of the compositions described herein.

[0032] [ 18 A kit containing DCFPyL precursors and reagents for use in the radiosynthesis of [F]DCFPyL is provided herein. In one embodiment, the kit comprises a DCFPyL precursor and a phosphate. In another embodiment, a catalyst comprising a DCFPyL precursor and tetrabutylammonium bicarbonate is used. A kit is provided.

[0033] In one embodiment of any one of the kits provided herein, the DCFPyL precursor is 5-(( (S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentan-2-yl) (I)ureido)-6-oxohexyl)carbamoyl)-N,N,N-trimethylpyridine-2-amini In one embodiment, the DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-5-(3-( (S)-1,5-Di-tert-butoxy-1,5-dioxopentan-2-yl)ureido)-6-oxohexyl (I)carbamoyl)-N,N,N-trimethylpyridin-2-aminium trifluoromethanesulfonate In another embodiment, the DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-5- (3-((S)-1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)ureido)-6-oxohe (xyl)carbamoyl)-N,N,N-trimethylpyridin-2-aminium trifluoroacetate It is a route.

[0034] In one embodiment of any one of the kits provided herein, the kit includes a DCFPyL precursor. and instructions for use for radiolabeling by any one of the methods provided herein. nothing.

[0035] In one embodiment of any one of the kits provided herein, the kit includes a DCFPyL precursor. The body and the above [ 18Includes QC Acceptance Specification (2016) for F]DCFPyL.

[0036] In certain embodiments of any one of the kits provided herein, the kit comprises one or more Cryptands, e.g., one or more Kryptofix® compounds In one embodiment of any one of the kits provided herein, the kit comprises , for example, packaged in a radiation-resistant container [ 18 F] fluoride ion, 18 F]Fluoride ion It further includes

[0037] The compositions and / or kits described herein can be used in imaging, e.g., diagnostic imaging. Thus, cells, organs, or tissues can be contacted with any of the compositions. Imaging methods are provided herein that allow for the detection of inflammatory bowel disease.

[0038] In another aspect, the method of administering any of the aspects of the composition to a subject comprises administering the composition of any of the aspects of the composition to a subject as described herein. For example, the method can be used for imaging and / or treating cancer. It may be used in the following cases.

[0039] Within the scope of the present invention are included: (i) a radiofluorination module (RFM) as described herein; or (ii) an apparatus including an ELIXYS automated radiochemical synthesizer; and (iii) any one of the apparatuses described herein. The method can be carried out using a radiofluorination module (RFM) or an ELIXYS automated radio and apparatuses including radiochemical synthesis apparatuses.

[0040] Certain aspects of the presently disclosed subject matter are described herein and are Aspects may be addressed in whole or in part depending on the subject matter disclosed. Other aspects are described below. As described herein, when illustrated in connection with the accompanying examples and figures, This will become clear as the description proceeds. [Brief explanation of the drawings]

[0041] Having described the subject matter disclosed herein in general terms, reference will now be made to the accompanying drawings, in which: However, they are not necessarily drawn to scale. [Figure 1] Figure 1 shows the synthesis of the DCFPyL precursor, 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carbamoyl)-N,N,N-trimethylpyridin-2-aminium trifluoromethanesulfonate (3). [Figure 2] Figure 2A and B show gradient HPLC of [18F]t-butyl protected DCFPyL (Figure 2A - radioactivity, Figure 2B - UV). [Figure 3] FIG. 3 shows the gradient UV HPLC of DCFPyL, the trimethylammonium precursor, and the fluorinated protected intermediate standard. [Figure 4] Figure 4 shows the gradient HPLC of the final crude [18F]DCFPyL before preparative purification. [Figure 5] FIG. 5 shows the radiosynthesis of [18F]DCFPyL. [Figure 6] Figures 6A and 6B show the radioactive (Figure 6A) and UV (Figure 6B) chromatograms of preparative HPLC of [18F]DCFPyL. [Figure 7]Figure 7A shows the QC chromatogram of [18F]DCFPyL. Mass of 0.0134 nmoles on support DCFPyL. Figure 7B shows the support-loaded chromatogram of [18F]DCFPyL. Upon addition of a standard solution of DCFPyL, the mass increases to 0.0384 nmoles. [Figure 8] Figure 8A and B show the gradient HPLC chromatograms of the final formulated [18F]DCFPyL. Figure 8B shows the UV trace of the final product in blue, with a blank injection of saline overlaid to show the gradient trace without the final product in gray.

[0042] This patent or application contains at least one drawing executed in color. Copies of the grayscale and color drawing(s) of this patent or patent application publication are available upon request and necessity. Any fee payment will be provided by the Authority. DETAILED DESCRIPTION OF THE INVENTION

[0043] The subject matter disclosed herein will be described in more detail below with reference to the accompanying drawings, in which: Some embodiments of the invention are shown, but not all embodiments are shown. Like numbers refer to like elements throughout. may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments should not be construed as limiting the scope of the present disclosure to any legal requirement to which it may be applicable. Indeed, the subject matter disclosed herein is provided to satisfy the requirements of Many modifications and other embodiments of the present invention may be made with the benefit of the teachings presented in the foregoing descriptions and the associated drawings. It will occur to one skilled in the art of the presently disclosed subject matter to The subject matter described is not limited to the particular embodiments disclosed and modifications and other embodiments are possible. It is to be understood that all such modifications are intended to fall within the scope of the appended claims.

[0044] The presently disclosed subject matter provides a method for radiochemical synthesis of prosthetic groups using an automated radiochemical synthesis module. by a multi-step synthesis involving reactive fluorination and coupling to urea [ 18 We present the preparation of [F]DCFPyL. The automated synthesis of this tracer involves the removal of the protecting ester group and purification. , two reactors, multiple separate synthesis steps using two precursors, 90 min synthesis time, and produces a low to moderate radiochemical yield of the final product.

[0045] This results in increased radiochemical yields, such as from a single precursor using automated synthesis [ 18 Improved methods for synthesizing [F]DCFPyL are provided herein. The composition was prepared using the method described above. 18 The method used is [F]DCFPyL. The method provided allows the production of a compound with high specific activity [ 18 The composition of [F]DCFPyL was obtained It has been found that this can be done.

[0046]

[0032] 18 The synthesis of [F]DCFPyL was compared with that of different halogen-based radioisotopes. It is also conceivable that this method could be applied to radiolabeling DCFPyL precursors with elements. can be.

[0047] [ 18 Synthesis of [F]DCFPyL In one embodiment, the PMSA inhibitor [18 F]DCFPyL is a single DCFPyL precursor, 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentane- 2-yl)ureido)-6-oxohexyl)carbamoyl)-N,N,N-trimethylpyridin-2-yl Radiofluorination of 3, structure shown in Figure 5 This may be synthesized by deprotection of the t-butyl group followed by purification.

[0048] Preferably, the methods provided include (i) forming a radiofluorinated DCPFPyL precursor. (ii) radiofluorination of a DCFPyL precursor containing an ester partial protecting group; In the mixture 18 [F]DCFPyL, (iii) deprotecting the ester moiety protecting group of [ 18 To provide [F]DCFPyL from the reaction mixture of step (ii) 18 and purifying the [F]DCFPyL. In [ 18 The radiofluorination and deprotection steps to form [F]DCFPyL were carried out in one reaction. This is done in a container or one pot.

[0049] In some embodiments, the DCFPyL precursor is a compound of formula (I) or a salt thereof: : [ka] ; Here, Q is a protecting group for the ester moiety that can be removed by treatment with phosphoric acid. As used herein, a "protecting group" refers to a regenerated functional group or group in a molecule. is a chemical group that can be selectively removed by readily available reagents that do not attack other functional groups. Suitable protecting groups can be selected, for example, from those described by Wutz et al. ("Greene's Protective Groups in in Organic Synthesis, 4th ed., Wiley-Interscience, 2007). For protection of the ester moiety, as described by Wutz et al. (pp. 533-643), In certain embodiments, the following protecting groups are used. Specific examples of protecting groups include, but are not limited to: However, benzyl, p-methoxybenzyl (PMB), tertiary butyl (tert-butyl or t- Butyl), Methoxymethyl (MOM), Methoxyethoxymethyl (MEM), Methylthiomethyl (MTM), tetrahydropyranyl (THP), tetrahydrofuranyl (THF), benzyloxy Dimethyl (BOM), trimethylsilyl (TMS), triethylsilyl (TES), t-butyldimethylsilyl tritylsilyl (TBDMS) and triphenylmethyl (trityl, Tr); and where L is a nucleophilic heteroaromatic substitution reaction [ 18 to form [F]DCFPyL , wherein the DCFPyL precursor is [ 18 F] a chemical moiety that allows it to combine with fluoride ions or In some embodiments, L may be a positively charged atom or group of atoms. In embodiments, L is tri(C1-C6 alkyl)ammonium (e.g., trimethylammonium). nium), and mineral acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, perchloric acid, nitric acid and sulfuric acid, and positively charged atoms or groups derived from organic acids, e.g., tartaric acid, thiazolinone, Trifluoroacetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, Succinic acid, methanesulfonic acid, trifluoromethanesulfonic acid and para-toluene a positively charged atom or group of atoms derived from an ene sulfonic acid; Preferably, chlorides, bromides, perchlorates, sulfonates, nitrates, phosphates, and thiamins. with a counterion selected from trifluoromethanesulfonate, more preferably trifluoromethanesulfonate with a methylolmethansulfonate counterion.

[0050] In one embodiment, the DCFPyL precursor has structure (I) above, where Q is tert-butyl. It has.

[0051] In one embodiment, the DCFPyL precursor is a compound in which L is N + (CH3)3 or trimethylammonium It has the above structure (I), which is a hydroxyl group salt.

[0052] In one embodiment, the DCFPyL precursor is a compound in which Q is tert-butyl and L is N + (C H3)3 or the trimethylammonium salt of structure (I) above.

[0053] As used herein, the term "deprotecting" refers to the removal of a carbonyl group from a This refers to the removal of the protecting group on the ester moiety to be converted into a hydroxy group.

[0054] Radiofluorination Module The radiofluorination module is a system for radiofluorinating compounds. To minimize radiation exposure, the modules are automated and remotely controlled to For example, a radiofluorination module may contain several different sub-modules. Each sub-module may contain a 18 F]DCFPyL In some embodiments, the radiofluorination module is configured to perform the steps of the synthesis method. The dry [ 18 First sub-method for preparing [F]fluoride ion Module, DCFPyL precursor [ 18 The second sub-module responsible for the radiosynthesis of [F]DCFPyL and from the radioactive synthesis reaction mixture [ 18 The third sub-module purifies [F]DCFPyL. Each sub-module may include, for example, a tube and / or a pump. In some embodiments, the radiofluorinated module may be operably connected. The module is 18 [F] fluoride ions. And that [ 18 Dry [F]fluoride ion for radiofluorination 18 [F] fluoride ion The first sub-module to prepare may receive:

[0055] In one embodiment, the radiofluorination module is configured to monitor the reaction conditions. including on-line sensors (e.g., sensors for temperature, pressure, flow rate, and radioactivity). In one embodiment, the radiofluorination module may further comprise: To monitor the quality of the reaction product after each step, the sub-modules are installed downstream or upstream. It may be further configured to include a line analyzer.

[0056] In some embodiments,18 [F]DCFPyL was added to a custom-made radiofluorination module ( In one embodiment, the RFM hardware comprises a heating block, Two syringe pumps, e.g., two Tecan Carvro syringe pumps, e.g. Multi-port caps, such as those configured for standard v-vials, and valves In one embodiment, the RFM further comprises a temperature heating cavity. In one embodiment, the temperature heating cavity is provided by a microwave cavity. In one embodiment, the v-vial is a 5 mL v-vial.

[0057] The RFM is installed on a laptop running Labview Real-Time software. National Instruments Compact Field Port connected to a computer National Instruments Compact Fieldpoint source module e) may be used to control the radioactive fluorination module. The software is National Instruments LabVIEW Professional and LabV Built on the IEW Real-Time platform, automated RFM and control software Exemplary configurations of the algorithm are described by Ravert et al. (Ravert et al., 2014) and Ravert et al. (Ravert et al., 2014). rt et al., 2015), and such configurations are incorporated herein by reference in their entirety. In one embodiment, the hardware-software system , [ 18The complete procedure, including the recovery of [F]fluoride and the injection of the reaction mixture into a semi-preparative HPLC, was performed. In some embodiments, RFM can be configured for partial automation. be.

[0058] In some embodiments, 18 [F]DCFPyL was synthesized using an ELIXYS automated radiochemical synthesizer (Sophie Bai Sciences, Inc., Culver City, CA (Sophie The compound is synthesized using the method described in Lazarie et al., Biosciences, Inc., Culver City, CA (Lazarie et al., 2014). In an embodiment, only one of the three reactors in the ELIXYS is [ 18 To synthesize [F]DCFPyL Used for this purpose.

[0059] Generally, components that come into contact with or are exposed to the reactants, reaction intermediates, or products (e.g., The surfaces of the composite module (lubricant or tube) are inert and / or non-reactive. During radiosynthesis, any reactants, reaction intermediates, or products of the material (e.g., made of or containing (acid or alkali and / or materials that minimize adhesion) In one embodiment, fluorine contamination in radiosynthesis To minimize contamination, components (e.g., valves or piping) and / or composite modules Some or all of the surfaces of the core are fluorine-free. "Free of fluorine" means that the fluorine atom or fluoride ion is 0.01% or less ( For example, 0.005% or less, 0.001% or less, 0.0001% or less, or 0% is included. In some embodiments, components (e.g., valves or tubes) and / or composite modules Part or all of the surface of the module is made of fluoropolymer such as polytetrafluoroethylene. Does not include.

[0060] In some embodiments, a flow path is provided that minimizes transfer losses and transfer times from upright, small-volume vessels. Use a body pathway design.

[0061] DCFPyL precursor, [ 18 F]fluoride, and [ 18 Synthesis of [F]fluoride standards In one embodiment, the DCFPyL precursor is a compound A having structure (II) and a compound B having structure (III). It may be synthesized by acylation reaction with compound B or its salt having the formula: [ka] wherein Q in compound A having structural formula (II) is a compound obtained by the treatment of phosphoric acid as defined above. is a protecting group for the ester moiety removable by wherein L in the compound B having the structural formula (III) or a salt thereof is as defined above. It is a leaving group.

[0062] In certain embodiments, compound A having structural formula (II) is: [ka] ; and where Q is a tert-butyl group.

[0063] In certain embodiments, compound B having structural formula (III) is: [ka] It has a counterion as defined above, preferably trifluoromethanesulfonate. do.

[0064] In one embodiment, the DCFPyL precursor (3,5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5- Di-tert-butoxy-1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carba Moyl-N,N,N-trimethylpyridin-2-aminium trifluoromethanesulfonate ) as shown in Figure 1, trimethylammonium nicotinate triflate It can be synthesized by coupling salt (2) with uriedo compound (1). Compound (1) in dichloromethane is mixed with triethylamine (TEA) and compound (2). After incubation at room temperature, the product was dried and eluted with acetonitrile and dimethyl Forms a semi-solid in ether. In some embodiments, applying a vacuum to dry The DCFPyL precursor is purified using a C-18 column (e.g., C-18 Sep-Pak Vac). The counter ion of the DCFPyL precursor may be exchanged during purification. As a result, in one embodiment, the purified DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-2-methyl-2-methyl-1-propanol). 5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)ureido)-6-o (N,N,N-trimethylpyridin-2-aminium trifluoro)-(1,2-hexyl)carbamoyl In one embodiment, the purified DCFPyL precursor is 5-(((S )-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentan-2-yl )Ureido)-6-oxohexyl)carbamoyl)-N,N,N-trimethylpyridine-2-aminyl In one embodiment, the fraction is lyophilized to a white solid. It may form a body.

[0065] In one embodiment, oxygen-18 enriched water is prepared by dissolving niobium in a niobium-based solution. General Electric Medical Systems (GEMS, Uppsala, Sweden) The high yield of the PETtrace cyclotron at Gems (GEMS, Uppsala, Sweden) 18 F] fluoride and irradiating the target with a proton beam, 18 F] Fluoride It can generate things.

[0066] ((2S)-2-[[(1S)-1-carboxy-5-[(6-fluoroanylpyridine-3-carbonyl)-amino ]pentyl]-carbamoylamino]pentanedioic acid, DCFPyL standard, ((2S)-2-[[(1S)-1-t-butyl ethylcarboxylate-5-[(6-fluoroanylpyridine-3-carbonyl)-amino]-pentyl ]-Carbamoyl-amino]-di-t-butyl pentanedioate, fluorinated protected Intermediate standard, 2-[3-[1-t-butylcarboxylate-(5-aminopentyl)]-uriedo ) ]-di-t-butyl pentanedioate (1) formate and N, N, N-trimethyl-5-((2, 3 , 5,6-Tetrafluorophenoxy)carbonyl)-pyridin-2-aminium trifluoro Methods for synthesizing methanesulfonate (2) are provided herein. Exemplary methods are described below. These studies have been published (Chen et al., 2011; Banerjee et al., 2008; Olberg et al., 2010), and such The method description is incorporated herein by reference in its entirety.

[0067] Radiofluorination, deprotection and purification Methods are provided for radiofluorinating the precursors provided herein. In one embodiment, the method comprises: (i) In the cartridge 18 F] scavenging fluoride ions; (ii) Captured in the cartridge [ 18 Tetrabutyltin to release [F] fluoride ions Dissolving ammonium base salts (e.g., tetrabutylammonium bicarbonate (TBABC)) Elution from the cartridge with liquid; (iii) The above [ 18 F] drying the eluate containing fluoride ions; (iv) The dried [ 18 [F] fluoride ion in an organic or aprotic solvent (e.g. adding a solution of DCFPyL precursor in acetonitrile; and (v) heating the mixed solution of step (iv); Includes.

[0068] In one embodiment, the cartridge is treated with an organic solvent, such as acetonitrile, after elution. It may be rinsed with a medium.

[0069] In one embodiment, all chemicals and components are first placed in the RFM or ELIXYS synthesis cassette. Then, fill the [ 18 [F] fluoride ions were extracted using Chromafix 30-PS-HCO3 solid phase extraction (SPEX). E) Cartridge (ABX, Radeberg, Germany) (Chromafix 30-PS-HCO3 Solid Phase anion exchangers such as Streptomyces Extraction (SPE) cartridges (ABX GmbH, Radeberg, Germany). and in one embodiment, washing the cartridge with high purity water. High purity water is commercially available, for example, from Fluka. High-purity water has a temperature of approximately 10°C at 25°C. -8 S / cm (e.g., 5.5 × 10 -8 Electrical conductivity (or The reciprocal electrical resistivity is about 10 MΩ cm, e.g., 18 MΩ cm. In some embodiments, the volume of water used for preconditioning is 0.5 to 2 ml. L (e.g., 1 mL). 18 O] Recover water for reuse. In some embodiments, the captured [ 18 The resin cartridge with [F] fluoride ions is then and eluted with a solution of tetrabutylammonium bicarbonate (TBABC). When using RFM as described above, the volume should be about 500 to about 700 μL (e.g., 600 μL). ) is used for elution. In one embodiment, when using the ELIXYS system, In this case, use about 200 to about 400 μL (e.g., 300 μL) of TBABC for elution. In embodiments, other bases (e.g., Kryptofix® 2.2.2 In some embodiments, the solvent is potassium bicarbonate or potassium acetate. The vial used to collect the exudate was cleaned with dilute nitric acid and washed with high-purity water (e.g., HPLC water). and dry overnight at 80°C.

[0070] In one embodiment, the eluted [ 18 F] fluoride ion-containing solution After rinsing the cartridge with acetonitrile, it was placed in a standard temperature heating block. Drying is carried out at between about 80°C and about 150°C (e.g., 110°C) using a controlled nitrogen flow. In some embodiments, the heating time is from about 50 seconds to about 300 seconds (e.g., 150 seconds), from about 250 to about 400 msec. In one embodiment, a nitrogen flow of between 100 and 200 mL / min (e.g., 325 mL / min) is used. Eluted from the 18 [F] fluoride ion-containing solutions were diluted with anhydrous acetonitrile one or more times. For example, in one embodiment, the azeotropic drying is carried out by successive addition and removal of Add acetonitrile more than once (e.g., two, three, or four times) to further dry the sample. Heat for about 50 to 300 seconds each time. For example, add acetonitrile twice separately. (250 μL each) for 150 and 180 seconds, respectively, or 90 and 180 seconds, respectively. In one embodiment, the acetonitrile is heated under vacuum and nitrogen. In some embodiments, the vial may be heated under a stream of compressed air. Cool to a temperature of 40°C to 60°C (e.g., 45°C or 50°C). Use an air flow of 5 to 10 L / min (e.g., 6 L / min).

[0071] A solution of the DCFPyL precursor (e.g., 3 in FIG. 5) in acetonitrile is then added to the Dried [ 18 It may be added to a reaction vial containing [F] fluoride ions. In embodiments, the solution is stirred for about 2 minutes to about 10 minutes (e.g., 5 minutes or 6 minutes), about Heating is performed at a temperature between 30° C. and about 70° C. (e.g., 45 or 50° C.). The solution is applied for about 20 to about 200 seconds (e.g., 20, 30, 60, 100, 150, or 200 seconds). The microwave irradiation is performed at about 40 W to about 60 W (e.g., 40 W, 50 W, or 60 W).

[0072] For the deprotection step, about 300 to about 400 μL (e.g., 350 μL) of phosphoric acid (60 to 100%) is added. 90%, e.g., 75% or 85%, or an acid having a pKa of 1.8 to 2.5 (e.g., 1.8, 1.9, 2.0, 2.1, 2.12, 2.2, 2.3, 2.4, 2.5), for example, by cooling the reaction mixture. In one embodiment, the vial is heated for about 2 minutes to about 10 minutes (e.g., 10 minutes). The reaction is then maintained at about 30°C to about 55°C (e.g., 45°C) for a period of time, e.g., 6 minutes. and buffering the reaction at a pH of about 2 to about 2.5. Buffering is achieved by the addition of sodium hydroxide and sodium dihydrogen phosphate buffer. Exemplary concentrations and volumes of drugs are as follows: (2 M, 2 mL) and sodium dihydrogen phosphate buffer (10 mM, pH 2.1, 1 mL).

[0073] [ 18 Standard techniques known in the art can be applied to purify [F]DCFPyL. In one embodiment, the crude reaction mixture is loaded onto a C18 column and purified by adding methanol and phosphoric acid. For example, the crude reaction mixture is eluted with a 15:85 mixture of methanol and sodium dihydrogen ether. elution may occur in a mixture of 0.01 N sodium dihydrogen phosphate (pH 2.1). In this embodiment, 18 [F]DCFPyL was collected in the HPLC water reservoir. The Sep-pak Plus Long cartridge was flushed with nitrogen and rinsed with HPLC water. The radiotracer product was then diluted with absolute ethanol followed by a 0.2 μm sterile filter. In one embodiment, the product is eluted with sterile saline. In one embodiment, the recovery and / or elution is carried out by an ascorbic acid solution. It is carried out in the presence of sodium benzoate.

[0074] In some embodiments, 18 The synthesis method for [F]DCFPyL conforms to all standard USP Chapters <823> Receiving USP Chapter 1 <823> Acceptance Testing Criteria In some embodiments, the radioactive compounds described herein are used to generate large mCi quantities. The method requires at least 20 mCi of 18 F]DCFPyL, e.g., at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least At least 100, at least 150, at least 200, at least 300, at least 400, at least and 500, at least 600 mCi [ 18 F]DCFPyL.

[0075] HPLC analysis For quality control HPLC analysis, various known analytical chromatographies known in the art are used. In some embodiments, the system may be used. 18 Chemical and activation of [F]DCFPyL The chemical properties may be measured using an Agilent 1260 Infinity System. Quaternary pump, HiP ALS autosampler, and NaI radioactivity detector Add a Bioscan Flow-Count interface with Max-L set at 264 nm An exemplary Agilent 1260 Infinity equipped with a DAD UV detector with a light flow cell. The system configuration is similar to that of Ravert et al. J. Label Compt. Radiopharm 2014, 57:695, J. Label Co mpt. Radiopharm 2015, 58:180), which is incorporated herein by reference in its entirety. In one embodiment, the Agilent OpenLAB Chromatography Data System A chromatograph is used to collect and analyze chromatographic data. One exemplary combination of service conditions is as follows: Atlantis T3 C18 5 μm 4.6×150 m m (Waters Corp., Milford, MA) ) in acetonitrile (MeCN):triethylamine (TEA) / phosphate buffer (pH 3.2) Elution was performed with a 10:90 mixture at a flow rate of 2 mL / min, and UV was set at 264 nm. In some cases, the following compound is used as a standard for HPLC analysis: ((2S)-2-[[(1 S)-1-Carboxy-5-[(6-fluoroanylpyridine-3-carbonyl)-amino]pentyl]-carboxamide bamoylamino]pentanedioic acid, DCFPyL standard, ((2S)-2-[[(1S)-1-t-butylcarboxyle 5-[(6-fluoroanylpyridine-3-carbonyl)-amino]-pentyl]-carbamoyl- Amino]-di-t-butyl pentanedioate, a fluorinated protected intermediate standard.

[0076] composition The product composition obtained from any one of the methods provided herein may be any of the compositions provided herein. Such compositions may be used alone or in combination as required for their intended use. It may be used in combination with other ingredients or compounds.

[0077] In one embodiment, the 18 [F]DCFPyL should be at least 10 Ci / μmole, e.g., at least At least 20, at least 30, at least 40, at least 50, at least 60, at least 70 , at least 80, at least 90, at least 100, at least 110, at least 120 , at least 130, at least 140, at least 150 Ci / μmole, or greater In some embodiments, the [ 18 F]DCFPyL ranges from ca. It may have an average specific activity of approximately 150 Ci / µmole.

[0078] In some embodiments, the compositions described herein may be at a concentration of about 400 ppm or less, e.g., 300 ppm or less. ppm or less, 200 ppm or less, 100 ppm or less, 50 ppm or less, 25 ppm or less, 10 ppm or less, 5 p pm or less, 1 ppm or less, or even lower concentrations of acetonitrile. In this case, the compositions described herein do not contain acetonitrile.

[0079] In some embodiments, the compositions described herein contain less than about 3,000 ppm, e.g., less than 2,000 ppm. m or less, 1,000 ppm or less, 500 ppm or less, 250 ppm or less, 100 ppm or less, 50 ppm or less, Contains methanol at a concentration of 10 ppm or less, 1 ppm or less, or even lower. In the present invention, the compositions described herein contain methanol at a concentration of between about 0 and 50 ppm.

[0080] In certain embodiments, the compositions described herein contain one or more cryptands, e.g., Does not contain more than one Kryptofix® compound.

[0081] In some embodiments, the compositions described herein do not include t-butanol.

[0082] In some embodiments, the compositions described herein do not include triethylamine.

[0083] In certain embodiments, the compositions described herein comprise a cryptand, e.g., a cryptophile. Kryptofix® compound, t-butanol, and triethylamine Also does not include.

[0084] In some embodiments, the compositions described herein are at least 95% or more, e.g., at least 96%, at least 97%, at least 98%, at least 99%, or at most 100% %of [ 18 F]DCFPyL with radiochemical purity.

[0085] Device Also, any of the compounds that may be used to practice any one of the methods provided herein. Synthesis modules are provided herein. Exemplary modules include those described in the Examples. Preferred embodiments include RFMs such as those described herein, and the ELIXYS correction system provided herein. In an embodiment, the module includes any one of the methods provided herein. In another preferred embodiment, the module comprises the elements described above and in the examples. It is of the type.

[0086] How to use contacting a cell, organ or tissue with, or administering to, a subject an effective amount of a compound provided herein imaging one or more cells, organs, or tissues, including administering a compound that Methods are also provided herein. In some embodiments, one or more organs or tissues include , prostate tissue, kidney tissue, brain tissue, vascular tissue or tumor tissue.

[0087] In one embodiment, the imaging method involves imaging by targeting PSMA. In another embodiment, the imaging method is suitable for imaging cancer, tumor or In a further embodiment, the cancer is of the eye or ocular. Cancer, rectal cancer, colon cancer, cervical cancer, prostate cancer, breast and bladder cancer, oral cancer, Benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, and prostate cancer cancer of the testicles, kidney, brain (e.g., glioma), throat, and skin Chromoma, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell Cancer and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, lymphoma, neurofibromatosis The tumor is selected from the group consisting of rheumatoid arthritis, tuberous sclerosis, hemangioma, and lymphangiogenesis.

[0088] The imaging methods provided herein are useful for imaging any physiological process in which PSMA is involved. In one embodiment, the imaging method is suitable for imaging a feature. The method is suitable for identifying tissues or target areas that express high levels of PSMA. Applications include imaging of glutamate neurotransmission, presynaptic glutamatergic neurotransmission, PSMA-expressing malignant tumors or cancers, prostate cancer (including metastatic prostate cancer) and angiogenesis Solid tumors express PSMA during angiogenesis. The methods and compositions described herein can be used to treat tumors, such as lung, renal, glioblastoma, pancreatic, bladder, sarcoma, melanoma, and the like. Imaging solid tumors including esophageal, breast, colon, germ cell, pheochromocytoma, and gastric tumors. PSMA is present in the peri- and intratumoral regions of various malignancies. It is frequently expressed in the endothelial cells of capillaries in the brain, and is used to image such malignancies. The methods and compositions provided may also be used to perform, for example, uterine Endometrium, schwannoma, and Barrett's esophagus. Certain benign lesions and tissues, including leukocytes, can be imaged according to the methods and compositions provided. There is a saying.

[0089] The provided methods and compositions for imaging angiogenesis are directed to imaging the manner in which angiogenesis occurs. Suitable for use in imaging a variety of diseases and disorders. Examples include tumors, collagen vascular disease, cancer, stroke, and vascular malformations. These include ocular malformation and retinopathy. The methods and compositions are also suitable for use in diagnosing and monitoring the development of normal tissue. are.

[0090] In certain embodiments of any one of the methods or compositions provided herein, the radiolabel The compositions of compounds have high specific activities, such as the levels of specific activity described herein.

[0091] In certain embodiments of any one of the methods or compositions provided herein, the radioactive The labeled compounds are then analyzed by positron emission tomography (PET) or positron emission tomography / computed tomography (PET). Detected by PET / CT. Images are obtained by measuring the difference in the spatial distribution of contrast agents that accumulate in a certain area. The spatial distribution may be generated by any means suitable for the particular label, e.g. For example, measurements can be made using a gamma camera, PET device, PET / CT device, etc. The extent of accumulation of the imaging agent can be determined using known methods for quantifying radioactive emissions. It can be quantified.

[0092] Generally, for imaging, a detectably effective amount of the compositions provided herein is administered. According to the present invention, a "detectably effective amount" may be administered to a subject. "Effective amount" means the amount of time required to obtain acceptable images using clinically available equipment. A detectable and effective amount of the compositions provided herein is defined as an amount sufficient to: It may be administered in two or more injections. The detectably effective amount depends on the degree of sensitivity of the individual, Varies with factors such as body age, sex and weight, individual-specific responses, and dosimetry. The detectably effective amount may also vary depending on factors related to the device and film. Optimization of such factors is well within the level of ordinary skill in the art. The amount of contrast agent and the duration of the imaging examination depend on the radionuclide used to label the agent; the patient's weight, the nature and severity of the condition being treated, the nature of the therapeutic treatment the patient is receiving; It depends on the patient's unique response. Ultimately, the attending physician will decide the dosage and image to be administered to each individual patient. The duration of the aging test may be determined.

[0093] In certain embodiments of any one of the methods or compositions provided herein, the subject is in humans, rats, mice, cats, dogs, horses, sheep, cattle, monkeys, birds, or amphibians. In another embodiment of any one of the methods or compositions provided herein, the cells The cells may be in vivo or in vitro. Typical examples of cells to which the compounds of the present invention can be administered include: The subject is a mammal, such as a primate or human. For veterinary use, a wide range of species are suitable. Subjects of this type include, for example, domestic animals such as cattle, sheep, goats, cows, and pigs. Livestock; poultry such as chickens, ducks, geese, turkeys, etc.; and domestic animals, especially For diagnostic or research purposes, a wide variety of Mammals include, for example, rodents (e.g., mice, rats, hamsters), rabbits, and primates. Suitable subjects include pigs, including inbred and inbred pigs. For in vitro applications, such as research applications, body fluids and cell samples from any of the above subjects may be used, for example, Suitable for use with blood, urine or tissue samples, for example from humans.

[0094] Another embodiment of the present invention is to administer a therapeutically effective amount of the compositions provided herein, preferably and a method and composition for treating tumors comprising administering to a subject a therapeutically effective amount of In one embodiment, the tumor cells are, for example, prostate tumor cells or metastatic prostate tumor cells. In another embodiment, the tumor may be a PSMA-expressing tumor, such as a PSMA-expressing tumor. Treatment can be achieved by targeting adjacent or nearby cells that are involved in the treatment. For example, The present invention can target vascular cells that are involved in tumor angiogenesis. The methods and compositions described herein are useful in treating tumors, such as lung, renal, glioblastoma, pancreatic, bladder, sarcoma, and melanoma. , breast, colon, germ cell, pheochromocytoma, esophagus and stomach (or as described herein or To treat solid tumors, including any other cancer or tumor known to those skilled in the art, It may also be used in the treatment of, for example, the endometrium, schwannoma, and Certain benign lesions and tissues, including Barrett's esophagus, are treated with the provided method. and compositions.

[0095] A "therapeutically effective amount" is an amount effective for therapeutic purposes. Generally, with respect to a composition for administration to a subject, the amount of the active ingredient in the composition is an amount effective for the administration of the active ingredient to the subject. Effective doses refer to the amount of a composition that produces one or more desired responses in a given dose. A suitable amount is any amount of the compositions provided herein that produces such a desired response. This amount may be for in vitro or in vivo purposes. For in vivo purposes, The amounts listed are those that a clinician would consider may have clinical benefit to a subject. Such subjects may include any one of the subjects described herein. A therapeutically effective amount is an amount that produces a desired therapeutic endpoint or a desired therapeutic result. Achieving any of the above may be accomplished by routine methods, including the amounts of the compositions provided herein. can be monitored by

[0096] A therapeutically effective amount will be determined based on the specific subject being treated; the severity of the condition, disease or disorder; Individual patient parameters, including, for example, age, physical condition, size and weight; duration of treatment; the nature of concomitant therapy (if any); the specific route of administration and the knowledge and expertise of the healthcare professional These factors are well known and routinely performed by those skilled in the art. The maximum dose may be determined according to sound medical judgment. It is generally preferable to use the safest dose. The vendor has the right to refuse to provide the patient with a lower risk of developing a condition that may be due to medical, psychological, or virtually any other reason. It will be appreciated that a larger or more tolerated dose may be required.

[0097] kit Kits containing any one of the compositions provided herein are also provided. In one embodiment, the kit comprises a package containing a pharmaceutically acceptable carrier and the composition of the present invention. In another embodiment, the kit comprises a pharmaceutical composition as provided herein. The present invention provides compounds and reagents necessary to carry out any one of the methods provided. In any one of the kit embodiments, the kit comprises: any one of the compositions provided herein; or any one of the methods of use provided herein. Instructions for the final radiolabeled compound as provided herein in the method of use and further includes an indicia including at least one of:

[0098] Kits containing DCFPyL precursors and reagents for use in radiofluorination are provided herein. In one embodiment, the kit comprises a DCFPyL precursor and a phosphate. In one embodiment, the kit comprises a DCFPyL precursor and tetrabutylammonium carbonate. In one embodiment, the kit comprises one or more chlorine salts, and optionally phosphate. Liptands, such as Kryptofix® (e.g., Crypto In one embodiment, the kit does not contain Kryptofix® 2.2.2. The 18 [F] fluoride ion (e.g., packaged in a radiation-resistant container) 18 F] Fluoride It may further contain cations).

[0099] In one embodiment, the kit comprises a compound as described herein in combination with a pharmaceutically acceptable carrier. The composition of any one of the provided kits includes a solution. In the case of a lyophilized form, the kit may be in the form of a liquid or a lyophilized form. Reconstitution with sterile and physiologically acceptable solutions, such as water, saline, buffered saline, etc. In another embodiment, the compound may be in solution or lyophilized form. In some cases, the kit may contain other additives, such as NaCl, silicate, phosphate buffer, ascorbic acid, gelatin, etc. Stabilizers may optionally be included, such as antiseptic acid and the like.

[0100] A "pharmaceutically acceptable carrier" is a carrier that is sterile, p [Eta] refers to a biocompatible solution that takes into account isotonicity, stability, etc., and any and all solvents. , diluents (e.g., sterile saline, sodium chloride injection, Ringer's injection) Injection), Dextrose Injection, Dextrose and Sodium Chloride Injection, Lactic Acid Lactated Ringer's Injection and other aqueous buffers, etc.), dispersion media The pharmaceutically acceptable carrier may contain, for example, a carrier, a coating, an antibacterial and antifungal agent, an isotonic agent, etc. The carrier may also contain stabilizers, preservatives, antioxidants, or other additives well known to those skilled in the art. or other vehicles known in the art.

[0101] As used herein, the term "pharmaceutically acceptable salt" refers to a compound selected from the group consisting of hydroxybenzoates, ... "(e salts)" refers to compounds that are modified by making their non-toxic acid or base salts. Pharmaceutically acceptable salts refer to derivatives of the disclosed compounds. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; acidic residues such as carboxylic acids; The pharmaceutically acceptable salts include, for example, non-toxic, inert salts of the compounds of formula (I), (II), (III), (IV ... These include the conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from organic or synthetic acids. For example, conventional non-toxic acid salts include those containing inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfuric acid, Salts derived from acetic acid, propionic acid, succinic acid, etc. , glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, Pamoic acid, maltic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid Citric acid, mesylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluene sulfonate Sulfonic acid, methanesulfonic acid, ethane disulfonic acid, oxalic acid, isethionic acid, HOOC-(CH2) From organic acids such as n-COOH (wherein n is 0 to 4, including, for example, 0, 1, 2, 3, and 4). The pharmaceutically acceptable salts of the present invention include those prepared by conventional chemical methods. Generally, such salts can be synthesized from a parent compound that contains a basic or acidic moiety. The free acid forms of these compounds are reacted with a stoichiometric amount of a suitable base (e.g., Na, Ca, Mg, or is obtained by reacting with K (hydroxide, carbonate, bicarbonate, etc.) or in the free base form. These compounds can be prepared by reacting them with a stoichiometric amount of an appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or a mixture of the two. Typically, the reaction is carried out in ether, ethyl acetate, ethanol, isopropanol, or In practice, non-aqueous media such as acetonitrile are used. A list of additional suitable salts is provided in, e.g. For example, Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, East on, Pa., p. 1418 (1985). Use of a physiologically acceptable salt in any one of the methods or compositions provided. There is.

[0102] As used in this specification and the appended claims, the singular forms "a," "an," and "the" " includes plural referents unless the context clearly dictates otherwise. Long-standing patent law practice Accordingly, the terms "a," "an," and "the" are used in this application, including the claims. In the case of a subject, the term "one or more" refers to a subject. " refers to a plurality of subjects unless the context clearly indicates otherwise (e.g., subjects), including multiple subjects, etc.

[0103] Throughout this specification and claims, unless the context requires otherwise, the term "includes" "comprise", "comprises" and "comprising" are non-exclusive Similarly, the term "include" and its grammatical variations are used to refer to a list. Describing an item in the specification may identify other similar items that may replace or be added to the listed item. It is not intended to be limiting or to exclude.

[0104] In this specification and the appended claims, unless otherwise indicated, Amounts, sizes, dimensions, proportions, shapes, and formulations used within the scope of the request , parameters, percentages, quantities, properties and all other numbers that represent numerical values The word "about" is used to refer to any value, amount, or range, even if the term "about" does not explicitly appear in conjunction with the value, amount, or range. It should be understood that all instances are modified by the term "about." Accordingly, unless indicated to the contrary, the following specification and appended claims The numerical parameters set forth are not, and need not be, precise, and may be adjusted as desired. , approximately and / or may be greater or smaller, tolerances, conversion factors, rounding may reflect inputs, measurement errors, etc., and may be subject to the subject matter disclosed herein. Depending on the desired properties sought to be obtained, and reflecting other factors known to those skilled in the art. For example, referencing a value may also include variations of the value. When referring to a particular amount, the term "about" refers to a range, in some embodiments, of ±100% from the particular amount. %, in some embodiments ±50%, in some embodiments ±20%, in some embodiments ±10 %, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5 %, and in some embodiments ±0.1%, and such variations are within the scope of the present invention when practicing the disclosed methods. or for using the disclosed compositions.

[0105] Additionally, the term "about" when used in connection with one or more numerical values ​​or numerical ranges, and all such values ​​are to be understood as referring to, for example, Variations that fall within a range and that extend the boundaries above and below that range. etc. The recitation of a numerical range by endpoints includes all numbers subsumed within that range, e.g. For example, all integers and their fractions (e.g., writing 1 to 5 also means 1, 2, 3, 4 and 5 and fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, etc.) and ranges thereof Any range within the range is included. [Example]

[0106] The following examples are provided to guide those skilled in the art in practicing representative embodiments of the present subject matter. In light of this disclosure and the general level of skill of one of ordinary skill in the art, The examples are illustrative only and those skilled in the art will appreciate that many variations may be made without departing from the scope of the invention. It is understood that changes, modifications, and variations may be made thereto.

[0107] Example 1 [ 18 Radiopharmaceutical-grade synthesis of [F]DCFPyL Two different automated platforms with fully compliant quality control specifications On the form, click 18 The radioactive synthesis of [F]DCFPyL is described below. The radioactive tracer synthesis is Custom-made Radiofluorination Module (RFM) and Sophie Biosciences The RFM synthesis was performed on an ELIXYS automated radiochemical synthesizer from NIH Inc. The RFM synthesis was performed at the end of irradiation time (end-of-irradiation time). The synthesis was completed within an average of 66 minutes from the end of bombardment (EOB) to the end of synthesis (E The mean specific activity of EOS was 4.4 TBq / μmole (120 Ci / μmole) and the mean release rate of EOS was 30.9%. The ELIXYS synthesis yielded an average radiochemical yield of 87 The assay was completed within minutes, with an average specific activity of 2.2 TBq / μmole (59.3 Ci / μmole) and 19 Both synthesis modules met all standards of USP Chapter <82. 3> Acceptance Test Criteria (standard USP Chapter <823> acceptance testing criteria While fitting, large mCi amounts of [ 18 F]DCFPyL was generated.

[0108] In this example, a custom-built synthesis module was used in our laboratory. The automated synthesis is performed in both the m-build synthesis module and a commercially available radiochemistry module. [ 18 The improved synthesis of [F]DCFPyL was performed in a single reactor, in five operating steps, and in six steps. The synthesis time ranged from 5 to 87 minutes, resulting in increased radiochemical yields. Using the method, radiopharmaceutical production levels of synthesis can be achieved.

[0109] Experimental Method All chemicals and solvents were of ACS or HPLC purity and, except as noted, Igma-Aldrich Chemical Company (St. Louis, MO) or F Purchased through Isher Scientific (Waltham, MA) ((2S)-2-[[(1S)-1-carboxy-5-[(6-fluoroanylpyridine-3-carbonyl)- [amino]pentyl]-carbamoylamino]pentanedioic acid, DCFPyL standard, ((2S)-2-[[(1S)-1-t- Butylcarboxylate-5-[(6-fluoroanylpyridine-3-carbonyl)-amino]-pentyl [I]-carbamoyl-amino]-di-t-butyl pentanedioate, fluorinated protected The intermediate standard, 2-[3-[1-t-butylcarboxylate-(5-aminopentyl)]-uriedo do) ]-di-t-butyl pentanedioate (1) formate and N,N,N-trimethyl-5-((2, 3,5,6-Tetrafluorophenoxy)carbonyl)pyridin-2-aminium trifluoride Fluoromethanesulfonate (2) was synthesized as described (Chen et al., 2011; Banerjee et al., 2012). 008; Olberg et al., 2010). The description of such synthesis methods is incorporated in its entirety unless otherwise stated. is incorporated herein by reference.

[0110] The custom-made radiofluorination module (RFM) is used for microwave radiosynthesis. Module (Ravert et al., 2014), the description of which is incorporated herein by reference in its entirety. The microwave cavity was constructed and controlled in a similar manner to that described in the previous paper. Instead of a microwave cavity, a thermal heating cavity is used. ELIXYS (Sophie Biosciences, Culver City, California) was used. The Sofie Biosciences, Inc., Culver City, CA) module is commercially available. It is an automated multi-reactor radiosynthesizer (Lazari et al., 2014).

[0111] For routine quality control (QC) HPLC analysis, a quaternary pump is used. , a HiP ALS autosampler, and a NaI radioactivity detector (Eckert & Ziegler, Berlin, Germany). Add a Bioscan Flow-Count interface with a meter set at 264 nm. Agilent 1260 Infinity Systèmes incorporating a DAD UV detector with an ax-Light flow cell. Using STEM (Santa Clara, CA), 18 F]DCFP The chemical and radiochemical identity of yL was determined. Chromatographic data was acquired and analyzed. Analysis was performed using the Gilent OpenLAB Chromatography Data System (Rev. A.04.05). The following chromatographic conditions were used: Atlantis T3 C185 μm 4.6 × 150 mm (Wall Waters Corp., Milford, MA column Acetonitrile (MeCN):triethylamine (TEA) / phosphate buffer (pH 3.2) 10:9 The column was eluted with a mixture of 0 at a flow rate of 2 mL / min, and the UV was set at 264 nm.

[0112] To investigate the possibility of residual lipophilic starting materials, To do this, gradient HPLC analysis was performed using the same Agilent HPLC system and initially with solvent A at a flow rate of 1 ml / min. Waters A equilibrated with 10:90 MeCN:TEA / phosphate buffer (pH 3.2) The analysis was performed using a Tlantis dC 185 μm 2.1 × 100 mm column. Solvent A (100%) was injected. The solvent B ( The concentration of MeCN (100% MeCN) was increased linearly from 0 to 85%.

[0113] [ 18 Analysis of the residual solvent levels of [F]DCFPyL batches was performed using an Agilent 7890A gas chromatograph. Agilent OpenLAB Chromatography Data Acquisition for graphing, data acquisition and analysis System, and WAX (polyethylene glycol phase: USP G16, G20) 30 meters, 0.25 mm ID, 0.25 μm film column.

[0114] 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentasiloxane (2-methyl-2-yl)ureido)-6-oxohexyl)carbamoyl)-N,N,N-trimethylpyridine-2- Synthesis of aminium trifluoromethanesulfonate (3) 1 (0.303 g, 0.57 mmole) was dissolved in 6 mL of dichloromethane. TEA and 2 (0.272 g, 0.57 mmole) were added. The reaction mixture was stirred at room temperature for 1 h. The solvent was then removed under a stream of N2, and the mixture was then dried under vacuum. The mixture was kept at room temperature for 30 minutes until it became semi-solid. The ether layer was removed and the semi-solid was redissolved in acetonitrile. Ether was added with stirring to produce a semi-solid. The mixture was kept at room temperature for 30 minutes. The ether layer was removed. The semi-solid was dried under vacuum and diluted with acetonitrile / water (1:9 The mixture was purified in a C-18 Sep-Pak Vac 35 cc (Waters) using a 5:5 (v:v) dilution of 1000 ppm from 1000 ppm. The fractions were collected and lyophilized to give a white solid (0.322 g, 71%). 1H-NMR (500 MHz, MeOD) δ 9.04 (d, J = 2.3 Hz, 1H), 8.54 (dd, J1= 2.3 Hz, J2= 8.7 Hz, 1H), 8.10 (d, J = 8.7 Hz, 1H), 4.17 (m, 2H), 3.68 (s, 9H), 3.43 (m, 2H), 2.32 (m, 2H), 2.03 (m, 1 13C-NMR (500 MHz, MeOD) δ 1 74.0, 173.9, 173.6, 166.0, 160.1, 159.7, 149.5, 141.4, 134.3, 121.9 (q, J = 317. 9 Hz), 115.8, 83.0, 82.8, 81.9, 56.0, 54.9, 54.3, 41.1, 33.5, 32.6, 29.9, 29.1, 28.5, 28.4, 24.2. Elemental analysis: Calculated for C34H56F3N5O11S; C, 51.05; H, 7.06; N, 8.76 Found: C, 50.58; H, 6.95; N, 8.49. HR-MS calculated for C33H56N5O8+: 650.4123, found Value, 650.4138 [M]+.

[0115] [ 18 [F] fluoride production Oxygen-18 enriched water (98%, Huayi Isotopes, Jiangsu, China, approximately 2 mL) was added to the niobium body. General Electric Medical Systems (GEMS, Uppsala, Sweden) PETtrace Cycling System from General Electric Medical Systems (GEMS, Uppsala, Sweden) High yield of rothorone 18 [F] fluoride target, filled with 55 μA. proton beam for 30 minutes, 18 O(p,n) 18 Approximately 61 GBq (1.65 Ci) of F was released by nuclear reactions. Aqueous [ 18F] fluoride ion was generated.

[0116] Radiofluorination Module (RFM) was used [ 18 Radiosynthesis of [F]DCFPyL After loading all chemicals and components into the RFM, 18 [F] fluoride ion in 1 mL of high Chromafix 30-PS-HCO3SPE cart preconditioned by washing with purified water (Fluka) Ridge (ABX, Radeberg, Germany) (Chromafix 30-PS-HCO3SPE cartridge (ABX GmbH, Radeberg, Germany). 18 O] was recovered and reused. NI LabView, National Instruments, Austin, TX The resin cartridge was then filled with tetrabutylammonium bicarbonate (TBB) under the control of The mixture was eluted with a solution of ABC (600 μL, 0.075 M, ABX GmbH, Germany) and the resulting mixture was The solution was collected in a 5 mL reaction vial sealed with a multiport cap; Before the procedure, the cartridge was cleaned with dilute nitric acid, washed with HPLC water, and dried overnight at 80 °C. The edge was rinsed with MeCN (250 μL) and then heated in a standard temperature heating block under controlled nitrogen The solution was dried at 110°C for 150 seconds at a flow rate of 325 mL / min. 18 F] Fluoride To further dry the solution, MeCN (250 μL each) was added in two separate portions, The samples were heated for 150 and 180 seconds.

[0117] The vial was cooled to a temperature of 50°C using compressed air (flow of approximately 6 liters / min). The dried [ 18 Add the DCFPyL precursor to the reaction vial containing [F] fluoride ions. A solution of 3 (5 mg, 6.25 μmol) in MeCN (500 μL) was added. The solution was heated at 50 °C for 6 h. After this, without cooling, phosphoric acid (85%, 350 μL) was added. The solution was kept at 45 °C for another 6 min. Sodium hydroxide (2 M, 2 mL) and sodium dihydrogen phosphate were added. The reaction was stopped by adding a mixture of thorium buffer (10 mM, pH 2.1, 1 mL) and the reaction mixture The material was buffered to a pH of 2 to 2.5.

[0118] The crude reaction mixture was purified by elution with a Phenomenex Gemini C18 5 μm 10×150 mm column (Torrance, The 15:85 metabolite was injected remotely into a 10 mL / min tube in Torrance, CA. The column was eluted with a mixture of ethanol (MeOH):0.01 M sodium dihydrogen phosphate (pH 2.1). Add water (70 mL) to the reservoir. 18 [F]DCFPyL (RT = 18 min, k' = 15.4) was collected. The fractions were pushed onto a C-18 Sep-Pak Plus Long cartridge (Waters) with nitrogen. The cartridge was rinsed with HPLC water (10 mL). aqueous ethanol (1 mL), then filtered through a 0.2 μm Millipore FG filter (25 mm; Merck, Millipore FG filter (25 mm; Merck KGaA, Darmstadt, Germany) Pre-filled with sterile saline (4 mL) through a sterile saline (10 mL) tube (Germany). The cartridges were eluted into sterile product vials.

[0119] Using ELIXYS 18 Radiosynthesis of [F]DCFPyL After all chemicals and components are loaded into the ELIXYS synthesis cassette, 18 F] fluoride The ions were delivered to 5 mL V-vials in the dose calibrator. The sequence was started. The Chr pre-conditioned by washing with 1 mL of high-purity water (Fluka) was The Omafix 30-PS-HCO3 SPE cartridge (ABX, Germany) was filled with the above [ 18 [F] fluoride ion Pressed with nitrogen. 18 The water was recovered and reused. The ridge was then removed with a solution of TBABC (300 μL, 0.075 M, ABX, Germany) using a glass stirring rod. The cartridge was rinsed with MeCN (600 μL) and eluted into a 5 mL V-vial. The solution was then dried at 110°C under vacuum and a nitrogen stream for 270 seconds while stirring. N (600 μL) was added in two separate times and incubated under vacuum and nitrogen flow for 90 and 105 seconds, respectively. It was heated for a while.

[0120] The vial was cooled to a temperature of 45° C. 18 F] Contains fluoride A solution of DCFPyL precursor (3) (5 mg, 6.25 μmol) in MeFP (500 μL) was added to the reaction vessel. The solution was heated at 50°C for 6 minutes while stirring. Phosphoric acid (85%, 350 μL) was added to the reaction mixture. The reaction vial was continued to be heated at 45°C with stirring for an additional 6 minutes. Two equal volumes of 2 mL of sodium hydroxide (2 M, 2 mL) and sodium dihydrogen phosphate (1 The reaction was stopped by adding a mixture of 100 mM HCl, pH 2.1, 1 mL with stirring, and the reaction mixture was stirred at 2 to 2.5°C. The reaction mixture was buffered to a pH of 0.05.

[0121] [ 18 Purification and formulation of [F]DCFPyL was the same as described for the RFM radiosynthesis above. It was the same.

[0122] Quality control procedures, visual inspection Using remote control equipment and appropriate radiation shielding (lead glass), 18 F]DCFPyL product The vials containing the solution were visually inspected under bright light. They were clear and colorless, with no foreign matter present. If so, the product meets this acceptance specification.

[0123] Radiochemical identity The control standard solution was injected into the analytical HPLC to ensure that the system conditions were suitable (see Table 1). Check retention time and mass (check for agreement with standard curve) To determine the radiochemical identity, 18 Final injection matrix aliquot of [F]DCFPyL A volume (50 μL) of the reference material was mixed with an aliquot of the reference standard solution. The reaction time was measured by a radiation detector. 18 Retention time of [F]DCFPyL If the offset between the two detector systems is properly calibrated, , this product met this acceptance specification.

[0124] radiochemical purity Using the same HPLC system as described for the radiochemical identity test, 18 DCF An appropriate volume of PyL (50 μL) was added to the uncorrected dead zone in the radioactivity detection system. The injection volume was chosen to avoid time loss (due to the main peak). 18 Activation of [F]DCFPyL The percent of biological purity is [ 18 The radioactivity associated with the [F]DCFPyL peak was analyzed by chromatogram analysis. The radiochemical purity was determined by dividing by the total activity assayed and multiplying by 100. If so, the product met the acceptance specifications. A sample of this is shown in Figure 7 (DCFPyL-RT = 7.6 min, k' = 8.7).

[0125] specific activity [ 18 The specific activity of [F]DCFPyL was determined by measuring the activity of a calibrated aliquot of [ 18 Assayed radioactivity of [F]DCFPyL ( The saturation (mCi / mL) at the end of synthesis was measured by HPLC-UV as interpreted from a standard mass calibration curve. The concentration was calculated by dividing the mass of DCFPyL by the mass concentration of the carrier (µmole of DCFPyL per mL). If the specific activity is 1000 mCi / µmole or greater, the product meets this acceptance specification. It was decided to add.

[0126] chemical purity At high specific activities, the use of simple UV peak ratios as an indicator of chemical purity is difficult to achieve. Typically, it becomes inappropriate as is smaller. 18 To successfully synthesize [F]DCFPyL, Greater than 99.5% of the starting precursors must be removed during synthesis; i.e., the final product matrix The amount of precursor or by-product remaining in the matrix may be less than 0.5%. All other UV-absorbing HPLC components not resulting from the HPLC reaction must also be below the same permitted residual precursor concentration. The same HPLC system as described for the radiochemical identity test must be used. hand,[ 18 The mass of the carrier of [F]DCFPyL was measured. The initial HPLC column void volume (VV) was After the void volume, all other UV peaks were summed and considered as by-products. If the mass concentration of these by-products is 1.5 μg / mL or less, the product meets this acceptance specification. It was decided to add.

[0127] Residual solvent analysis 1675 μL of absolute ethanol (6.7%), 12.7 μL of acetonitrile (400 ppm), and methanol An aliquot of a 25 mL HPLC water standard solution spiked with 94.7 μL of ethanol (3000 ppm) was analyzed. , system suitability was determined. 18 Aliquots of [F]DCFPyL product matrix were injected. and the residual solvent level was measured from the standard as well as from a single-point curve. The acetonitrile level was 400 ppm or less, and the methanol level was 0.1 ppm or less. If the level is less than 3000 ppm and the ethanol level is less than 10%, this product The product was to meet this acceptance specification (Intl. Conf. on Harmonisation of Tech. Requirements for Registration of Pharm. for Human Use, 1997).

[0128] pH A drop of [ 18 The final product matrix was analyzed using pH indicator paper (ColorPhast-Indicator Strips). Trip-pH 2-9; sensitivity 0.3 to 0.5 units, EMD Chemicals, Gibbstown, NY The color of the strip was compared to the indicator chart. If it is 8.5, the product meets this acceptance specification.

[0129] Sterile Filter Integrity Testing [ 18 F]DCFPyL Sterile microfiltrate derived from the terminal filtration step The filter was washed with 5 mL of absolute ethanol, kept wet, and a calibrated pressure gauge (Millipor The distal end of the filter was placed in a liquid reservoir and the gas pressure was applied. The pressure was slowly increased until a pressure of 13 psi or more was reached without any visible bubble flow. In this case, the integrity of the Millipore Millex FG filter is not guaranteed. It is intended to satisfy the needs of our customers.

[0130] Radionuclide identity [ 18 The radioactivity content (mCi) in an aliquot of the final product matrix [F]DCFPyL was determined as a function of time (0 minutes; A) and again 15 minutes later (B) using a Capintec CRC-15R Radioisotope Dose Calibrator (Radioisotope Dose Calibrator) (Ramsey, NJ) The half-life was calculated using the following formula: The calculated half-life was 105 to 115 minutes. If so, the product met this acceptance specification. T 1 / 2 =(4.495) / (log A - log B)

[0131] Radionuclide purity Using a suitable gamma spectrometer, take an appropriate time and place to obtain a gamma spectrum. An aliquot was injected and assayed. The gamma spectrum obtained was 18 F emission The presence of any distinctive photoelectric peaks was analyzed. More than 99.5% of the total gamma ray emissions observed correspond to 0.511 and 1.022 MeV. If so, the product will meet this acceptance specification.

[0132] Endotoxin Test Charles River Laboratories EndoSafe Portable Testing System(Endosafe PTS Read er, Integrated Software Version 7.10, Service Pack 2.0, and printer; Wilmi Wilmington, MA 18 F]DCFPyL final product The endotoxin levels in the batches of thrombin were analyzed. If the endotoxin levels are less than or equal to 11 endotoxin units / mL, the product meets this acceptance specification. It was decided.

[0133] Sterility Test In the laminar flow hood, 18 [F]DCFPyL Final product Matri Samples of the mixture (approximately 100 μL each) were cultured in liquid thioglycollate medium and soybean casein digestion medium. The soil (Becton, Dickinson and Company) was added. The media were incubated at 32.5 ± 2.5°C and 22.5 ± 2.5°C, respectively, and the cells that showed positive growth were The turbidity was monitored daily. No growth was observed during the 14-day incubation period. If so, the product met this acceptance specification.

[0134] Retesting at the expiration date of the radiotracer To demonstrate the stability of the radiotracer product when stored under ambient conditions: A subset of the above acceptance tests was performed 360 min after the end of radiotracer synthesis. .

[0135] Results and Discussion [ 18 The manual radiosynthesis of [F]DCFPyL (Chen et al., 2011) was performed in an automated dual reactor. Synthesis platform (old nuclear interface "double FDG synthesis module" (an ol Suitable for routine use with the Nuclear Interface “Double FDG Synthesis Module”) As part of the initial evaluation of radiotracer synthesis for human use, The final radiotracer product ( A moderate amount of 2.3 GBq (62 mCi average) was produced. Table 1 shows the initial acceptance specifications and the product. The results of the evaluation are presented. Over time, this procedure was carried out over a two-year period. Over 100 radiochemical syntheses, there was a small variation of approximately 3% in the mean specific activity at the end of the synthesis. Possible radiochemical yields (not corrected for decay, estimated average [ 18 [F] Fluoride target yield (based on the previous data), was often lower.

[0136] [Table 1] QC Acceptance Specification: 18 Original synthesis of [F]DCFPyL and new characterization in RFM and ELIXYS modules (All tests except yield, specific activity and filter integrity were performed at 360 min.) Repeat testing was conducted at the expiration date and the results were consistent with those performed at EOS.)

[0137] Previous methods have yielded variable yields with low to moderate specific activity. The radiochemical yield and specific activity were significantly increased. 18 A synthetic method for [F]DCFPyL is provided and previously reported. Meets or exceeds all QC criteria established for larger radiopharmaceutical-level radiation. To enable the production of radioactive tracers, precursors, alternative synthesis platforms (e.g., custom-made systems in the laboratory), (including the development of new methods), and modifications of reaction and purification conditions were established.

[0138] A new precursor was synthesized: a protected trimethylammonium precursor (3) (Figure 1). By successfully synthesizing chemical compounds, 18 Progress towards the synthesis of a radioactive tracer of [F]DCFPyL The precursor, 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di- ... rt-Butoxy-1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carbamoyl )-N,N,N-trimethylpyridin-2-aminium trifluoromethanesulfonate (3) , trimethylammonium nicotinate triflate salt (2) and ureido compound It was synthesized in reasonable yield from coupling with uriedo compound (1).

[0139] Radiofluorination of the trimethylammonium precursor (3) at 50 °C was carried out using the base TBABC. This produced a clean reaction profile with high yields. (Kryptofix® 2.2.2) These typically gave lower yields. Increasing the amount of precursor did not increase the radioactive fluoride yield. Increasing the amount from 11.25 to 22.5 to 45 μmol increased the yield from 10% to 30%. Using gradient HPLC (conditions described in the Experimental Methods section), radioactive fluorinated The yield of the protected intermediate was determined to be 81% (Figure 2, RT = 8.9 min). As a control, the trimethylammonium precursor and the non-radioactive fluorinated protected A UV trace of the intermediate ( Chen et al., 2011 ) is overlaid ( Figure 3 ).

[0140] After radiofluorination, deprotection of the ester moiety was attempted with a number of different acids, but the results were The results were low yields and by-products were formed. butyl groups in reasonable yields (Li et al., 2006) without forming major radiochemical by-products. The final crude deprotected product was purified by gradient HPLC (R Analysis at 200 rpm (T = 2.0 min) revealed no radioactive fluorinated protected intermediates or The product was free of trimethylammonium precursors and in good yield (Figure 4).

[0141] We focused on a platform suitable for automating synthesis. Tam-made RFM (α7-nicotinic ligand, high specific activity [ 18 F]ASEM, to create The microwave synthesis module was adapted with thermal heating ( Ravert et al., 2015) was constructed. Fluorine-free components (valves, tubing, etc.), transfer from upright small volume reagent tubes A few changes, such as designing the fluid path to minimize transport losses and transfer times, make it easy to A computer-controlled synthesis device (National Instruments) was used. The radiofluorination module was used in the NIRS 8000 series (using NIRS 80 ... The software used to control the controller is National Instruments L Built on the abVIEW Professional and LabVIEW Real-Time platforms The hardware and software interface is provided by National Instruments. This was performed using the compactRio embedded controller. The NI NI LabVIEW Professional and NI NI Real-Time platforms are End-user interface with National Instruments' embedded controllers The end user can modify the instrument subroutines from the library and The user of the radiofluoridation module shall Custom design and program interfaces.

[0142] Radiochemical synthesis was transferred to the Sophie Biosciences ELIXYS module. provides fluid pathways and connects one or more of the three reactors and cassettes in the synthesis sequence. The reactor system uses interchangeable cassettes to allow any combination of reactors to be used. This synthesis requires a single cassette and reactor. This software provides a simple and easy method for developing methods or adapting existing syntheses. The ELIXYS has an interface that can be easily changed. Potential sources of fluorine contamination are is not strictly excluded, but 18 F]DCFPyL was obtained in reasonable yield with high specific activity. To shorten the reaction time, less TBABC solution was used, which resulted in a somewhat lower final yield. Uniquely integrated robotics for reagent transfer and mobile reactors The synthesis time for ELIXYS was longer than that for RFM.

[0143] Custom-made RFM and Sophie ELIXYS radiosynthesis modules 18 DCF The final radiosynthesis scheme of PyL is outlined in Figure 5. The amount of TBABC aqueous solution added (R (600 μL for FM vs. 300 μL for ELIXYS), whereas ELIXYS requires reaction dilution prior to HPLC purification. Adding liquid in portions vs. adding a larger volume in one go with RFM, except , radiochemical synthesis is the same process.

[0144] In both modules, the [ 18 [F] Fluoride ion Chromafix 30-PS-HCO3 The TBABC solution was then eluted from the cartridge, followed by heating and additional Azeotropically dried with acetonitrile. The tolyl solution was added to a reaction vial and the vial was heated. Phosphoric acid was added and heated to form t- The butyl protecting group was removed. 18 The pH of the [F]DCFPyL solution was adjusted by adding sodium hydroxide and phosphoric acid dihydrate. The pH was adjusted to 2 to 2.5 by adding sodium hydroxide buffer and mixing thoroughly. Injection into the rum produced a typical chromatogram, as shown in Figure 6. 18 F]D The CFPyL peak was collected in a water reservoir and subjected to automated solid-phase extraction (SPE) preparation, followed by 1 mL of ethanol. and normal saline to obtain 14 mL of the final product solution.

[0145] The final sterile solution obtained from the RFM radiosynthesis contained 4.4±0.3 TBq / μmole (120±9 with a mean specific activity of 20.8 ± 3.0 Ci / µmole EOS (uncorrected for decay). .4 GBq (562±91 mCi; n = 3) [ 18 F]DCFPyL. The average EOS non-decay corrected yield is calculated by the average synthesis time At 66 minutes, the final sterile solution obtained from the ELIXYS radiosynthesis was 30.9 ± 3.0%. 2.2 ± 0.5 TBq / µmole (59.3 ± 12.4 Ci / µmole EOS, not corrected for decay) ) with a mean specific activity of 13.8 ± 7.4 TBq (372 ± 199 mCi; n = 3) [ 18 F]DCFPyL The average EOS non-decay yield was The corrected yield was 19.4±7.8% with an average synthesis time of 87 minutes. Both platforms had uncorrected specific activities of 193 GBq / µmole at EOS in approximately 90 minutes. (5.2 Ci / μmole), with an average of 2.3 GBq (62 mCi) [ 18 The original polysaccharides that produce [F]DCFPyL Compared to stepwise synthesis, the mCi yield and specific activity were significantly improved.

[0146] Both radioactive synthesis modules were fabricated using the methods disclosed herein. [ 18 Complete QC data for the first three validation runs for [F]DCFPyL are summarized in Table 1. These newly reported results meet or exceed all previously published acceptance specifications. The only thing that changed was that the final specific activity of the radiotracer product was significantly Significantly improved radiotracer quality (as indicated by the increased width) Made [ 18 The results were a significant improvement over the previously used 2 [F]DCFPyL. The type of solvent no longer exists in the synthesis and is now known as Kryptofix (registered trademark). Trademark) 2.2.2 is no longer in use and therefore the final acceptance limits and acceptance specifications There is no need for a

[0147] Typical QC chromatograms and chemical identities for chemical and radiochemical purity determination Co-injection ("spiked authentic") chromatography for sexual purposes The formula is shown in Figure 7. The average goodness of fit (R 2 ) with seven mass levels of non-radioactive DCFP yL (6 replicate injections per mass level) and the mass and The measurement of the support mass was determined from a calibration curve showing the relationship between the UV absorbance prepared over a range of 67 nmoles. The lower limit (0.0046 nmole) was established as the limit of quantitation at a signal-to-noise ratio of 6:1. The limit of detection was determined to be approximately half that amount (0.0023 nmole) (USP(1225)). . [ 18 Gradient HPLC of the final formulated solution of [F]DCFPyL revealed that the radioactive fluorinated The absence of protected intermediates or trimethylammonium precursors was demonstrated (Figure 8). ).

[0148] Adapting the manual synthesis to an automated synthesis platform is a 10-step process ( Fluoride capture, fluoride release and drying, reaction with the first precursor, intermediate purification of the first precursor Preparation of the tert-butyl-protected trifluoroacetic acid followed by reaction with a second precursor, evaporation of the reaction solvent, and release of the tert-butyl-protected trifluoroacetic acid. Deprotection of radioactively fluorinated intermediates, acid removal, buffering for preparative chromatography Such adapted syntheses include: A module with two separate reactors was used. In the synthesis described here, preparative H PLC is a method for preparing a single precursor without the need for solvent evaporation or acid removal prior to purification. The procedure is partly due to the use of a diamine and the deprotection of the penultimate product. The number of process steps can be reduced. The inherent release of radioactivity during solution transfer, evaporation, and intermediate purification with the added advantage of no loss of radioactivity and a shorter reaction time resulting in less radioactive decay. This synthesis shows a higher radiochemical yield at EOS compared to other syntheses.

[0149] lastly,[ 18 To demonstrate the feasibility of large-scale (multi-curie) production of [F]DCFPyL, The synthesis using the RFM module yielded 213 GBq (5.77 Ci) of the first [ 18 F] from fluoride 83. 6 Gbq (2.26 Ci) [ 18 [F]DCFPyL was produced in a yield of 39.2% (EOS). Specific activity (EOS) The radiochemical purity was 97.6%. In S, all other QC data were within the acceptance specifications set in Table 1. However, this large-scale radiotracer product will be retested at the prescribed six-hour expiration date. The product showed a radiochemical purity of only 51.1%. The validation was 1.6 GBq / mL (44 mCi / mL), and with a 6-hour expiration, it was 5.7 GBq / mL (153 mCi / mL), which was confirmed to meet all QC acceptance specifications, while significant release was observed. Decomposition of the radioactive tracer was observed. Sodium ascorbate was added to the saline used to elute the product from the column. Large-scale synthesis using the RFM module, which adds 222 GBq (6.0 Ci), was performed with the first [ 18 F ] 67.7 Gbq (1.83 Ci) from fluoride [ 18 F]DCFPyL was produced in a yield of 30.5% (EOS). The specific activity (EOS) was 5.9 TBq / μmole (159.3 Ci / μmole), and the radiochemical purity was At EOS, all other QC data were within the acceptance specifications set in Table 1. This was within 4.5 GBq / mL (122 mCi / mL) including added sodium ascorbate. Solution 18 The radiochemical purity of [F]DCFPyL at 3, 4, and 6 hours after EOS was 98.8%, respectively. %, 98.5%, and 98.2%.

[0150] summary Customized, high-yield, ultra-high specific activity radiofluorination synthesis modules were used. , PSMA inhibitors [ 18 F]DCFPYL etc. 18 Designing and constructing to produce [F] radiotracer The specific activity of PET radiotracers may be related to mass-dependent receptor localization or pharmacological toxicity. Critical quality control release criteria for concern ) Ultra-high specific activity can extend the shelf life of radioactive tracers, A single manufactured batch of tracer allows for many more PET studies. The synthesis module allows for the automated and semi-automated synthesis of radiotracers used in PET imaging. Automated radiochemical synthesis is now possible. There are currently over 10 commercial radiochemical synthesis modules available. What makes this chemistry module unique is that it can be used to The quality of the radiotracer produced is a key factor in the custom radiofluorination module of the present invention. 10 to 200 times greater specific activity than any of the commercial synthetic modules available today It has.

[0151] The radioactive fluorination module comprises: 18 F]Radiotracer products:[ 18 F]DCFPYL, [ 18 F]A SEM, 18 F]T807, and [ 18 F]AZAN, including but not limited to, can be used Cut.

[0152] [ 18 [F]DCFPyL was prepared with very high specific activity and moderate to high radiochemical yield. All regulatory acceptance specifications are documented and met for each batch of radiotracer produced. This synthesis using one of these automated modules allows for the Up to six PET / CT scans per day (per dose) with injections and imaging at short intervals 9 or 10 mCi) 18 [F]DCFPyL radiotracer products readily available will be done.

[0153] <References> All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. All publications, patent applications, patents, and related art documents are indicative of the level of ordinary skill in the art to which the subject matter disclosed herein pertains. Each individual publication, patent application, patent, and other reference is hereby incorporated by reference. All such modifications and variations are expressly incorporated herein by reference to the same extent as if specifically and individually indicated to be incorporated by reference. Many patent applications, patents and other references are cited herein. However, such references are not intended to be limiting unless any of these documents are of general general utility in the art. It is understood that no admission is made that the invention forms part of the general knowledge of the present invention. In the event of a conflict between references, the present specification (and any other references that may be incorporated herein) shall prevail. Unless otherwise specified, the standard practice in this specification The technically accepted meanings of terms are used. Use standard abbreviations.

[0154] 1)B. Li, M. Berliner, R. Buzon, CKF Chiu, ST Colgan, T. Kaneko, N. Keene, W. Kissel, KR Leeman, B. Marquez, R. Morris, L. Newell, S. Wunderwald, M. Wit t, J. Weaver, Z. Zhang, Z. Zhang, J. Org. Chem, 2006, 71, 9045. 2)DE Olberg, JM Arukee, D. Grace, OK Hjelstuen, M. Solbakken, GM Kindb erg, A. Cuthbertson, J Med Chem, 2010, 53, 1732. 3)HT Ravert, DP Holt, RF Dannals, A microwave radiosynthesis of the 4-[ 1 8F]-fluorobenzyltriphenylphosphonium ion, J Label Compds Radiopharm, 2014, 57, 6 95. 4)H.T. Ravert, D.P. Holt, Y. Gao, A.G. Horti, R.F. Dannals, Microwave-assisted radiosynthesis of [ 18 F]ASEM, a radiolabeled α7-nicotinic acetylcholine recepto r antagonist, J Label Compds Radiopharm, 2015, 58, 180. 5)K. Raisa, PET Radiochemistry Automation: State of the Art and Future Trends in 18F-nucleophilic Fluorination, Current Organic Chemistry, Volume 17, Number 1 9, October 2013, pp. 2097-2107(11). 6)International Conference on Harmonisation of Technical Requirements for Regi stration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite Guideline I mpurities: Guideline for Residual Solvents, 1997. 7)M. Dietlein, C. Kobe, G. Kuhnert, A. Stockter, T. Fischer, K. Schomacker, M. Schmidt, F. Dietlein, BD Zlatoposkly, P. Krapf, R. Richarz, S. Neubauer, A.D rzezga, B. Neumaier, Mol Imaging Biol, 2015, 17, 575. 8)M. Lazari, J. Collins, B. Shin, M. Farhoud, D. Yeh, B. Maraglia, FT Chin, DA Nathanson, M. Moore, RM van Dam, J Nucl Med Tech, 2014, 42, 1. 9)R. Seigel, J. Ma, Z. Zhou, A. Jemal, CA Cancer J Clin 2014, 64, 9. 10)RC Mease, CL Dusich, CA Foss, HT Ravert, RF Dannals, J. Seidel, A. Prideaux, JJ Fox, G. Sgouros, AP Kozikowski, MG Pomper, Clin. Can. Res. 2008, 14, 3036. 11)SR Banerjee, CA Foss, M. Castanares, RC Mease, Y. Byun, JJ Fox,J. Hilton, SE Lupold, AP Kozikowski, MG Pomper, J. Med. Chem, 2008, 51, 4504 . 12)T. Maurer, M. Eiber, M. Schwaiger, JE Gschwend, Nature Rev. / Urology 2016 , 13, 226. 13) US Pharmacopeia Chapter <823> Radiopharmaceuticals for Positron Emission Tomography - Compounding. USP 32-NF29, 2009. 14)USP (1225) “Validation of Compendial Methods”. 15) V. Bouvet, M. Wuest, HS Jans, N. Janzen, AR Genady, JF Valliant, F. Benard, F. Wuest, Eur J Nucl Med Mol Imaging Research, 2016, 6, 40. 16)Y. Chen, M. Pullambhatia, CA Foss, Y. Byun, S. Nimmagadda, S. Senthamizc helvan, G. Sgouros, RC Mease, MG Pomper, Clin. Can. Res. 2011, 17, 7645. 17) Z. Szabo, E. Mena, SP Rowe, D. Plyku, R. Nidal, MA Eisenberger, ES A ntonarakis, H. Fan, RF Dannals, Y. Chin, RC Mease, M. Vranesic, A. Bhatnagar. , G. Sgouros, SY Cho, MG Pomper, Mol Imaging Biol, 2015, 17, 565.

[0155] The foregoing subject matter has been described in some detail by way of illustration and example for clarity of understanding. However, it will be apparent to those skilled in the art that certain changes and modifications may be made within the scope of the appended claims. It will be understood that

Claims

1. 2-(3{1-carboxy-5-[(6-[ 18 F]fluoro-pyridine-3-carbonyl)-amino]-pentyl} -ureido)-pentanedioic acid ([ 18 1. A method for the synthesis of a compound (F]DCFPyL), said method comprising: (i) To form the radiofluorinated DCPFPyL precursor, D containing an ester moiety protecting group was added. Radiofluorination of the CFPyL precursor; (ii) In the reaction mixture 18 Step (i) of radiofluorinating D to form [F]DCFPyL deprotecting the ester moiety protecting group of the CPFPyL precursor with phosphoric acid; and (iii) [ 18 from the reaction mixture of step (ii) to provide [F]DCFPyL 18 F]DCFPyL To purify; A method comprising:

2. The protecting group is benzyl, p-methoxybenzyl, tert-butyl, methoxymethyl, methoxy Thiethoxymethyl, methylthiomethyl, tetrahydropyranyl, tetrahydrofuranyl, Benzyloxymethyl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl 2. The method of claim 1, wherein the methyl group is selected from the group consisting of methyl methyl benzoate (TBDMS) and triphenylmethyl benzoate (TBDMS).

3. 3. The process according to claim 1 or 2, wherein steps (i) and (ii) are carried out in one reactor. 。

4. The synthesis was performed using a heating block, two syringe pumps, a multi-port cap, and Use a radiofluorination module (RFM) containing a reagent addition vial with a valve. The method according to any one of claims 1 to 3, wherein the method is automated by

5. The method of claim 4 , wherein the RFM further comprises a temperature heating cavity.

6. 4. The method of claim 1, wherein the synthesis is automated by using an automated radiochemical synthesizer.

10. The method according to any one of the preceding claims.

7. 10. The method of claim 4, wherein the RFM or the automated radiochemical synthesizer components are fluorine-free.

6. The method according to any one of the preceding claims.

8. The DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy) -1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carbamoyl)-N,N,N- 8. The compound according to claim 1, which is a trimethylpyridin-2-aminium compound. method.

9. The DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy) N,N,N-di-1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carbamoyl -trimethylpyridin-2-aminium trifluoromethanesulfonate, claim 8 The method described below.

10. The DCFPyL precursor is 5-(((S)-6-(tert-butoxy)-5-(3-((S)-1,5-di-tert-butoxy) -1,5-dioxopentan-2-yl)ureido)-6-oxohexyl)carbamoyl)-N,N,N- 9. The compound according to claim 8, which is trimethylpyridin-2-aminium trifluoroacetate. method.

11. The DCFPyL precursor is 【Chemical 1】 11. The method of claim 9 or 10, wherein the compound is synthesized according to the formula:

12. The DCFPyL precursor is selected from the group consisting of: N,N,N-trimethyl-5-((2,3,5,6-tetrafluorophenoxy)carbonyl)-pyridine- 2-Aminium trifluoromethanesulfonate and 2-[3-[1-t-butylcarboxylate-( 5-aminopentyl)]-uriedo)]-di-t-butyl pentanedioate ng, The method of claim 9 or 10, wherein the compound is synthesized by a method comprising:

13. The method further comprises: 【Chemistry 2】 or N, N, N-trimethyl-5-((2, 3, 5, 6-tetrafluorophenoxy)carbo (3-[1-tert-butyl)-pyridin-2-aminium trifluoromethanesulfonate carboxylate-(5-aminopentyl)]-uriedo)]-di-t-butyl pentanediol 10. The method of claim 8, further comprising synthesizing the DCFPyL precursor by coupling with a 13. The method of any one of 12.

14. Radiofluorination of the DCFPyL precursor comprises: 【Chemistry 3】 14. The method according to claim 1, wherein the method is carried out according to the formula:

15. Radiofluorinating said DCFPyL precursor comprises: (a) In the cartridge 18 F] scavenging fluoride ions; (b) Captured in the cartridge 18 Tetrabutyltin to release [F] fluoride ions eluting said cartridge with a solution of ammonium base salt; (c) dried [ 18 to form [F] fluoride 18 F] fluoride ion-containing solution drying the exudate; and (d) the dried [ 18 F] fluoride ion (compound (3)) 【Chemistry 4】 adding a solution of 14. The method of any one of claims 1 to 13, comprising:

16. 10. The method of claim 1, wherein the cartridge is an anion exchange chromatography cartridge.

5. The method according to claim 5.

17. The cartridge is placed in the cartridge. 18 [F] fluoride ions before capturing them.

17. The method of claim 15 or 16, wherein the preconditioning is carried out by washing with water.

18. Radiofluorination of the DCFPyL precursor yields (compound (3)) 【Chemistry 5】 and dried [ 18 [F] fluoride ion mixture solution, optionally at a temperature of from about 30° C. to about 70° C.

18. The method of any one of claims 15 to 17, further comprising heating at a temperature between 0.5 and 1.5°C. Law.

19. 20. The method of claim 18, wherein the heating is for about 2 minutes to about 10 minutes.

20. 20. The method of claim 18 or 19, wherein the heating is at about 50°C for about 6 minutes.

21. The heating reacts with the DCFPyL precursor and the dried [ 18 F] in a mixed solution with fluoride ions, Microwave irradiation at approximately 40 W to approximately 60 W for approximately 20 seconds to approximately 200 seconds was performed.

21. The method according to any one of claims 18 to 20,

22. The heating reacts with the DCFPyL precursor and the dried [ 18 F] in a mixed solution with fluoride ions, This is achieved by irradiating the cells with microwaves at approximately 50 W for approximately 30 to 150 seconds.

22. The method of claim 21 .

23. [ from step (c) 18 23. The method of claim 15, wherein the fluoride ions are dried. The method described in paragraph .

24. The above [ 18 [F] The eluate from step (c) containing fluoride ions is heated at a temperature between about 80°C and about 150°C.

24. The method of any one of claims 15 to 23, wherein the drying is carried out at 200°C.

25. 25. The method of claim 24, wherein the temperature is about 110°C.

26. The above [ 18 and drying the eluate of step (c) containing [F] fluoride ions under a stream of nitrogen.

26. The method of any one of 15 to 25.

27. 27. The method of claim 15, wherein the drying is performed for about 50 seconds to about 300 seconds. method.

28. 28. The method of claim 27, wherein the drying is performed for about 150 seconds.

29. For further drying, 3 CN was dried as described above. 18 F] fluoride ion, 29. The method of any one of claims 15 to 28.

30. The RFM or automated radiochemical synthesizer, or its reaction portion, is pre-treated with dilute nitric acid prior to synthesis.

30. The method of claim 4, wherein the product is cleaned, washed with water, and dried overnight at about 80°C. The method described below.

31. 30. The method of claim 1, wherein the deprotection with phosphoric acid is carried out at a temperature between about 30° C. and about 55° C.

10. The method according to any one of the preceding claims.

32. 32. The method of claim 31 , wherein the temperature is about 45° C.

33. 33. The method of claim 31 or 32, wherein the temperature is maintained for about 2 minutes to about 10 minutes.

34. After deprotection with phosphoric acid, the pH of the reaction mixture in step (ii) is adjusted to between about 2 and about 2.

5.

34. The method of any one of claims 1 to 33, further comprising adjusting the pH.

35. The pH of the reaction mixture was adjusted by adding sodium hydroxide and sodium dihydrogen phosphate buffer. The method of claim 34, wherein the adjustment is performed by

36. 36. The method of claim 1, wherein the purification is performed by liquid chromatography. How to do it.

37. 37. The method of claim 36, wherein the liquid chromatography comprises at least one C18 column. How to post.

38. [ 18 The solution containing [F]DCFPyL was eluted with the first eluent containing methanol and sodium dihydrogen phosphate.

38. The method of claims 36 to 37, wherein the solution is used to elute from the first C18 column.

39. The first elution solution contained methanol and sodium dihydrogen phosphate in a ratio of 15:85 at pH 2.

1.

39. The method of claim 38, wherein the methanol: 0.01 M sodium dihydrogen phosphate.

40. The above [ 18 The solution containing [F]DCFPyL was then applied to a second C18 column, and the first solution containing ethanol was added.

40. The method of claim 38 or 39, wherein the elution is performed with 2 elution solutions.

41. The purification step may be carried out by purifying the eluted [ 18 Filtration of [F]DCFPyL 41. The method of any one of claims 36 to 40, further comprising the step of:

42. 42. The method of claim 41, wherein the filtering is through a 0.2 μm sterile filter.

43. 43. The method of claim 41 or 42, wherein the filtration is into a sterile vial.

44. 44. The method of claim 43, wherein the sterile vial is pre-filled with sterile saline.

45. 45. Any one of claims 1 to 44, wherein the purification is carried out in the presence of sodium ascorbate. The method described below.

46. adding the sodium ascorbate to a collection reservoir, and / or 18 F]DCFPyL raw 46. ​​The method of claim 45, wherein the composition is added to the solution used to elute the compound.

47. After the purification 18 46. ​​The method of claim 1, wherein the yield of [F]DCFPyL is at least about 20 mCi.

10. The method according to any one of the preceding claims.

48. After the purification 18 48. The method of claim 47, wherein the yield of [F]DCFPyL is at least about 100 mCi. How to do it.

49. After the purification 18 48. The method of claim 47, wherein the yield of [F]DCFPyL is at least 400 mCi. method.

50. below: (i) There is [ 18 F] scavenging fluoride ions; (ii) the [ 18 [F] fluoride ion to release tetrahydrofuran. eluting the cartridge with a solution of butylammonium bicarbonate (TBABC); (iii) [ from step (ii) 18 drying the eluate containing [F] fluoride ions; (iv) [ from step (iii) 18 F] fluoride ion (compound (3)) 【Chemistry 6】 adding a solution of (v) heating the mixed solution of step (iv); A method for radiofluorinating a DCFPyL precursor, comprising:

51. 6. The cartridge of claim 5, wherein the cartridge is an anion exchange chromatography cartridge. The method according to claim 0.

52. The cartridge is placed in the cartridge. 18 [F] fluoride ions before capturing them.

52. The method of claim 50 or 51, wherein the preconditioning is performed by washing with water.

53. 53. The method of any one of claims 50 to 52, wherein the heating is between about 30°C and about 70°C. How to post.

54. 54. The method of claim 53, wherein the heating is for about 2 minutes to about 10 minutes.

55. 55. The method of claim 54, wherein the heating is at about 50°C for about 6 minutes.

56. The heating step comprises irradiating the mixed solution of step (iv) with microwaves at about 40 W to about 60 W for about 2 minutes.

53. The method of claim 50, wherein the irradiation is performed for 0 to about 200 seconds. The method described in paragraph .

57. The heating step comprises irradiating the mixed solution of step (iv) with microwaves at about 50 W for about 30 seconds to about 1 minute.

57. The method of claim 56, wherein the irradiation is for 50 seconds.

58. [ from step (iii) 18 [F] fluoride ions are dried.

10. The method according to claim 1.

59. The above [ 18 The eluate containing [F] fluoride ions was dried at a temperature between about 80 °C and about 150 °C.

59. The method of any one of claims 50 to 58,

60. 60. The method of claim 59, wherein the temperature is about 110°C.

61. [ from step (ii) 18 [F] The eluate containing fluoride ions is dried under a stream of nitrogen.

61. The method of any one of claims 50 to 60.

62. 62. The method of claim 50, wherein the drying is performed for about 50 seconds to about 300 seconds. method.

63. 63. The method of claim 62, wherein the drying is performed for about 150 seconds.

64. For further drying, 3 CN was dried as described above [ 18 F] fluoride ion, 64. The method of any one of claims 58 to 63.

65. Produced by the method of any one of claims 1 to 49 18 Contains [F]DCFPyL composition.

66. The above [ 18 F]DCFPyL at least about 50 Ci / μmole, or at least about 100 Ci / μmole 66. The composition of claim 65, having an average specific activity of

67. 67. The composition of claim 65 or 66, wherein the composition comprises acetonitrile at a concentration of about 400 ppm or less. The composition described.

68. 68. Any of claims 65 to 67, wherein the composition comprises methanol at a concentration of about 3000 ppm or less. The composition according to any one of claims 1 to 4.

69. 69. The composition of claim 68, wherein the composition comprises methanol at a concentration of about 50 ppm or less.

70. 70. Any one of claims 65 to 69, wherein the composition does not include one or more cryptands. The composition described in

71. 8. The method of claim 7, wherein the cryptand is Kryptofix®.

10. The composition according to claim 0.

72. 72. The composition of any one of claims 65 to 71, wherein the composition does not contain triethylamine. composition.

73. 73. The composition of any one of claims 65 to 72, wherein the composition does not include t-butanol. Finished product.

74. [ 18 65. The method of claim 65, wherein the radiochemical purity of the [F]DCFPyL ranges from about 95% to about 100%.

73. The composition according to any one of claims 1 to 73.

75. 75. A kit comprising the composition of any one of claims 65 to 74.

76. Kit containing DCFPyL precursor and phosphate.

77. Kit containing DCFPyL precursor and tetrabutylammonium bicarbonate.

78. 78. The kit of any one of claims 75 to 77, which does not include one or more cryptands. 。

79. 8. The method of claim 7, wherein the cryptand is Kryptofix®.

9. The kit according to claim 8.

80. [ 18 80. The kit of any one of claims 75 to 79, further comprising [F] fluoride ions.

81. Contacting a cell, organ or tissue with a composition according to any one of claims 65 to 74 and an imaging method comprising:

82. 74. A method of administering to a subject a composition described in any one of claims 65 to 73.

83. 83. The method of claim 82, wherein the method is for imaging.

84. 83. The method of claim 82, wherein the method is for treating cancer.

85. 50. A radiofluorination module (RFM) or automatic radiofluorination module according to any one of claims 4 to 49. Apparatus including a radiochemical synthesis apparatus.

86. A radiofluorination model capable of carrying out the method of any one of claims 1 to 64. Equipment including a radiochemical synthesis machine (RFM) or automated radiochemical synthesis machine.

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