阅读说明：本技术 放射性氟标记的苯基砜类化合物及制法和应用 (Radioactive fluorine labeled phenylsulfone compound, preparation method and application thereof ) 是由 王力 刘楠 陈跃 胡梅 周柳 龙睿铃 冯悦 杨丽萍 王长江 于 2021-07-28 设计创作，主要内容包括：本发明公开了放射性氟标记的苯基砜类化合物及制法和应用,属于放射性药物及核医学技术领域,解决现有技术中PET显像剂标记核素半衰期短,能有效利用时间有限,放化产率低,比活度不高,分离纯化困难的问题。本发明的放射性氟标记的苯基砜类化合物结构如式I所示,本发明方法包括：氨基酸单体或氨基酸酯或三肽与式II化合物于第一溶剂中反应,制得式I化合物。本发明还提供了该放射性氟标记的化合物在制备肿瘤诊断试剂中的应用。本发明化合物分子量小,具有适当的LogP,不剧烈改变半胱氨酸及其类似物氨基酸特性,前体不易过血脑屏障,对脑部肿瘤病变探测能力高,放化标记产率高,产物比活度高,产品半衰期适中,显像清晰、分辨率高。(The invention discloses a radioactive fluorine labeled phenyl sulfone compound, a preparation method and application thereof, belongs to the technical field of radiopharmaceuticals and nuclear medicine, and solves the problems that in the prior art, a PET imaging agent labeled nuclide has short half-life period, limited effective utilization time, low radiochemical yield, low specific activity and difficult separation and purification. The structure of the radioactive fluorine labeled phenylsulfone compound is shown as a formula I, and the method comprises the following steps: reaction of amino acid monomers or amino acid esters or tripeptides with formula IIThe compound is reacted in a first solvent to produce the compound of formula I. The invention also provides application of the radioactive fluorine labeled compound in preparing a tumor diagnostic reagent. The compound has small molecular weight, proper LogP, no drastic change in the amino acid characteristics of cysteine and its analogs, less blood brain barrier crossing of precursor, high detection capacity on brain tumor, high radiochemical labeling yield, high specific activity of the product, moderate half life of the product, clear development and high resolution.)

1. A radiofluorine-labeled phenylsulfone compound represented by the formula I,

wherein R is₁Is an amino acid monomer or an amino acid ester or a tripeptide, R₂Is [ 2 ]¹⁸F]-fluoroalkyl group or [ alpha ], [ alpha ] an¹⁸F]-fluoroalkoxy group or [ alpha ], [¹⁸F]-fluorine and derivatives thereof.

2. A compound of claim 1, wherein R is₁Comprises the following steps:

wherein n is 0, 1, 2, 3 or 4,

R₃＝H，-CH₃，-CH₂OH，-CH(CH₃)₃，-CH₂CH(CH₃)₂，-CH(CH₃)(CH₂)_mCH₃，-(CH₂)_mS(CH₂)_mCH₃，-(CH₂)_mCOOH，-(CH₂)_mNH₂，-Benzyl，-CH₂Ph，-(CH₂)_mNHC(NH)NH₂；

R₄＝H，-CH₃，-CH₃(CH₃)，-CH₂CH(CH₃)₂，-(CH₂)_xNH₂，-CH(CH₃)(CH₂)_xCH₃，-(CH₂)_xS(CH₂)_xCH₃，-(CH₂)_xCOOH，-(CH₂)_xCONH₂，-Benzyl，-CH₃Phe，-(CH₂)_xCOOCH₃；

m＝1～3，x＝1～3。

3. the compound according to claim 1 or 2,a is 0, 1, 2, 3 or 4.

4. The method for producing a radioactive-fluorine-labeled phenylsulfone compound according to any one of claims 1 to 3, comprising the steps of: reacting an amino acid monomer or amino acid ester or tripeptide with a compound of a formula II in a first solvent to obtain a compound of a formula I;

a is 0, 1, 2, 3 or 4.

5. The method of claim 4, further comprising the preparation of a compound of formula II: combining a compound of formula III with [, ]¹⁸F]Heating fluoride in a second solventReacting to obtain a compound of formula II:

wherein b is 0, 1, 2, 3 or 4.

6. The method according to claim 5, wherein the reaction temperature of the amino acid monomer or the amino acid ester or the tripeptide and the compound of the formula II is 20-40 ℃; preferably 30-40 ℃; more preferably 35 ℃;

or/and a compound of the formula III and¹⁸F]-the reaction temperature of the fluoride is 70-90 ℃; preferably 75-85 ℃; more preferably 80 deg.c.

7. The process of claim 5, wherein the molar ratio of amino acid monomer or amino acid ester or tripeptide to compound of formula II is: the molar weight of the amino acid monomer, the amino acid ester or the tripeptide is 0.003-0.01 mmol, and the compound of the formula II is 5-20 mCi;

or/and a compound of the formula III and¹⁸F]-molar ratio of fluoride: the compound of the formula III is 0.001 to 0.01mmol [, ], [ 2 ]¹⁸F]The fluoride is 5 to 20 mCi.

8. The method of claim 4, wherein the [ sic ], [ solution ]¹⁸F]-the fluoride comprises tetrabutylammonium fluoride, aluminium trifluoride or potassium fluoride;

the first solvent comprises methanol, dimethyl sulfoxide, tetrahydrofuran, ethanol, N-dimethylformamide, acetone, butanone or acetonitrile;

the second solvent comprises acetonitrile, dimethyl sulfoxide, tetrahydrofuran, ethanol, methanol, N-dimethylformamide, acetone or butanone.

9. The method according to claim 8, wherein the reaction system of the compound of formula II and the compound of formula III further comprises an alkaline buffer sodium carbonate buffer, a sodium borate buffer, a potassium bicarbonate buffer, a phosphate buffer, a HEPES buffer, a MES buffer, preferably a sodium borate buffer with a pH of 8.5.

10. Use of the radioactive fluorine-labeled phenylsulfone compound according to any one of claims 1 to 3 for the preparation of a tumor diagnostic reagent; preferably, the tumor is prostate cancer, lung adenocarcinoma, colorectal cancer, gastric cancer, colon cancer, liver cancer, breast cancer, glioma, pheochromocytoma, nasopharyngeal carcinoma, thyroid cancer or lymphoma.

Technical Field

The invention belongs to the technical field of radiopharmaceuticals and nuclear medicine, and particularly relates to a phenyl sulfone compound labeled by radioactive fluorine, and a preparation method and application thereof.

Background

Positron Emission Tomography (PET) imaging is one of the most advanced modern molecular imaging technologies, and has obvious advantages over other molecular imaging technologies in the aspects of early differential diagnosis of tumors, systemic imaging of late metastatic tumors, intervention of individualized treatment of tumors and dynamic imaging. By utilizing the characteristic of abnormal tumor metabolism, the PET can carry out sugar metabolism, lipid metabolism, nucleic acid metabolism and amino acid metabolism imaging on the tumor. Wherein, the [ alpha ], [ beta ] -a¹⁸F]Fluorodeoxyglucose (f)¹⁸F-FDG) is currently the most commonly used sugar-metabolizing PET imaging agent. But do not¹⁸F-FDG has the problems of poor specificity, non-uptake of certain tumor cells, uptake of inflammatory cells and the like, so that false positive or false negative results can appear in the differential diagnosis of tumors, and the clinical applicability of 18F-FDG PET is limited to a certain extent.

Amino acid metabolism imaging in metabolismPlays an important role in molecular imaging, and can make up for the differential diagnosis of neuropsychiatric diseases, tumors and cardiovascular and cerebrovascular diseases¹⁸Some deficiencies of F-FDG. In the aspect of PET drug research and development, an amino acid transporter is an important target for researching novel amino acid PET drugs, a plurality of amino acid imaging agents show good clinical application prospects, and an amino acid PET drug library is established to provide individual PET drugs for accurate tumor diagnosis and treatment. The amino acid metabolism imaging agent commonly used in clinic is L1-¹¹C]Methionine (A), (B), (C)¹¹C-Met)，[β-¹¹C]-L-dopa (¹¹C-DOPA)，(O-2-¹⁸F-fluoroethyl) -L-tyrosine (¹⁸F-FET)，[α-¹¹C-methyl]-L-tryptophan (C)¹¹C-AMT), but the imaging agents have the problems of short half-life of labeled nuclide, limited effective utilization time, low radiochemical yield, low specific activity, difficult separation and purification and the like due to the limitation of a labeling method.

Therefore, the provision of a PET imaging agent, which has high radiochemical label yield, high product specific activity, moderate product half-life, clear imaging and high resolution, can significantly improve the sensitivity and accuracy of tumor diagnosis, and is a problem to be solved by those skilled in the art.

Disclosure of Invention

One of the purposes of the invention is to provide a radioactive fluorine labeled phenylsulfone compound, which solves the problems of short half-life of labeled nuclide of a PET imaging agent, limited effective utilization time, low radiochemical yield, low specific activity and difficult separation and purification in the prior art.

Another object of the present invention is to provide a method for producing the radiofluorine-labeled phenylsulfone compound.

It is a further object of the present invention to provide use of the radiofluorine-labeled phenylsulfone compound.

In order to achieve the purpose, the invention adopts the technical scheme that:

the invention provides a radioactive fluorine labeled phenylsulfone compound shown as formula I,

wherein R is₁Is an amino acid monomer or an amino acid ester or a tripeptide, R₂Is [ 2 ]¹⁸F]-fluoroalkyl group or [ alpha ], [ alpha ] an¹⁸F]-fluoroalkoxy group or [ alpha ], [¹⁸F]-fluorine and derivatives thereof.

Preferably, the amino acid monomer, the amino acid ester and the tripeptide contain sulfydryl or/and amino.

In some embodiments of the invention, R is₁Comprises the following steps:

wherein n is 0, 1, 2, 3 or 4,

R₃＝H，-CH₃，-CH₂OH，-CH(CH₃)₃，-CH₂CH(CH₃)₂，-CH(CH₃)(CH₂)_mCH₃，-(CH₂)_mS(CH₂)_mCH₃，-(CH₂)_mCOOH，-(CH₂)_mNH₂，-Benzyl，-CH₂Ph，-(CH₂)_mNHC(NH)NH₂；

R₄＝H，-CH₃，-CH₃(CH₃)，-CH₂CH(CH₃)₂，-(CH₂)_xNH₂，-CH(CH₃)(CH₂)_xCH₃，-(CH₂)_xS(CH₂)_xCH₃，-(CH₂)_xCOOH，-(CH₂)_xCONH₂，-Benzyl，-CH₃Phe，-(CH₂)_xCOOCH₃；

m＝1～3，x＝1～3。

in some embodiments of the present invention, the first and second substrates,a is 0, 1, 2, 3 or 4.

The preparation method of the radioactive fluorine labeled phenylsulfone compound provided by the invention comprises the following steps: reacting an amino acid monomer or amino acid ester or tripeptide with a compound of a formula II in a first solvent to obtain a compound of a formula I;

a is 0, 1, 2, 3 or 4.

In some embodiments of the invention, the preparation method further comprises the preparation of a compound of formula II: combining a compound of formula III with [, ]¹⁸F]-heating fluoride in a second solvent to obtain a compound of formula II:

wherein b is 0, 1, 2, 3 or 4.

In some embodiments of the invention, the reaction temperature of the amino acid monomer or the amino acid ester or the tripeptide and the compound of the formula II is 20-40 ℃; preferably 30-40 ℃; more preferably 35 ℃;

or/and a compound of the formula III and¹⁸F]-the reaction temperature of the fluoride is 70-90 ℃; preferably 75-85 ℃; more preferably 80 deg.c.

In some embodiments of the invention, the molar ratio of amino acid monomer or amino acid ester or tripeptide to compound of formula II is: the molar weight of the amino acid monomer, the amino acid ester or the tripeptide is 0.003-0.01 mmol, and the compound of the formula II is 5-20 mCi.

Or/and a compound of the formula III and¹⁸F]-molar ratio of fluoride: the compound of the formula III is 0.001 to 0.01mmol [, ], [ 2 ]¹⁸F]Fluorination ofThe substance is 5 to 20 mCi.

In some embodiments of the invention, the [ alpha ], [ beta ] or a¹⁸F]-the fluoride comprises tetrabutylammonium fluoride, aluminium trifluoride or potassium fluoride;

the first solvent comprises methanol, dimethyl sulfoxide, tetrahydrofuran, ethanol, N-dimethylformamide, acetone, butanone or acetonitrile;

the second solvent comprises acetonitrile, dimethyl sulfoxide, tetrahydrofuran, ethanol, methanol, N-dimethylformamide, acetone or butanone;

in some embodiments of the present invention, the reaction system of the compound of formula ii and the compound of formula iii further comprises an alkaline buffer sodium carbonate buffer, a sodium borate buffer, a potassium bicarbonate buffer, a phosphate buffer, a HEPES buffer, and a MES buffer, preferably a sodium borate buffer with a pH of 8.5.

The application of the radioactive fluorine labeled phenylsulfone compound in preparing a tumor diagnosis reagent; preferably, the tumor is prostate cancer, lung adenocarcinoma, colorectal cancer, gastric cancer, colon cancer, liver cancer, breast cancer, glioma, pheochromocytoma, nasopharyngeal carcinoma, thyroid cancer or lymphoma.

Compared with the prior art, the invention has the following beneficial effects:

the invention creatively adopts amino acid monomer or amino acid ester or tripeptide as a mark framework and adopts¹⁸F]-fluoroalkyl group or [ alpha ], [ alpha ] an¹⁸F]-fluoroalkoxy as a marker group to give the compounds of the formula I according to the invention. The radioactive fluorine labeled compound of the invention has small molecular weight, proper LogP, small molecular weight, does not change the characteristics of cysteine and analogues amino acid violently, the precursor does not cross blood brain barrier easily, the detection capability to brain tumor lesion is high, the radioactive fluorine labeled compound has the characteristics of high radiochemical labeling yield, high product specific activity, moderate product half-life period, clear development, high resolution and simple and easy labeling method, and can be used for carrying out labeling on amino acid containing naked sulfydryl and amino and analogues thereof¹⁸F is marked and used as a diagnostic reagent of positron emission computed tomography of malignant tumors.

Drawings

FIG. 1 is a drawing of¹⁸A radioactive detector HPLC chart of F-FVSB;

FIG. 2 is¹⁸HPLC chart of the F-FVSB ultraviolet detector;

FIG. 3 is¹⁸A radioactive detector HPLC plot of F-FVSB- (L) cysteine;

FIG. 4 is a drawing of¹⁸HPLC chart of ultraviolet detector for F-FVSB- (L) cysteine;

FIG. 5 is a drawing of¹⁸A radioactive detector HPLC chart of F-FVSB-GSH;

FIG. 6 is a drawing of¹⁸An HPLC chart of an ultraviolet detector of F-FVSB-GSH;

FIG. 7 is¹⁸The result of the development experiment of F-FVSB- (L) cysteine;

FIG. 8 is a drawing of¹⁸Imaging experiment result graph of F-FVSB-GSH.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

Example 1

This example discloses compounds of formula II-1¹⁸The preparation method of the F-FVSB comprises the following steps: the reaction formula is as follows:

¹⁸F-FVSB marking step: taking 2mg of the compound of formula III-1, placing in a dry sealed bottle, adding 100. mu.L of acetonitrile, and adding dry fluorine [ 2 ]¹⁸F]TBAF (15mCi, 50. mu.L), heated at 80 ℃ for 20 min.

A purification step: after completion of the reaction, 200. mu.L of 5% acetic acid was added to the reaction system to dilute the mixture, and the mixture was filtered through a filter and then subjected to liquid phase separation.

HPLC conditions: mobile phase 80% ACN (0.1% TFA) flow rate: 1mL/min, and the chromatographic column comprises: eclipse Plus C18, 5 μm, 4.6 × 250 mm. The column temperature was 35 ℃.

¹⁸The HPLC chart of F-FVSB is shown in FIG. 1 and FIG. 2, in which the detector of FIG. 1 is a radioactivity detector, the detector of FIG. 2 is an ultraviolet detector, and the detection wavelength is 254 nm.

Collecting eluate with peak time of 8.6min, diluting the collected eluate with water until ACN content is below 5%, hanging on HLB solid phase extraction column, washing HLB column with 10mL water, eluting with 200 μ L methanol, and collecting eluate to obtain¹⁸F-FVSB。

Example 2

This example discloses compounds of formula I-1¹⁸Preparation of F-FVSB- (L) cysteine, its reaction formula is:

taking 1mg of (L) cysteine, putting the cysteine into a 2ml centrifuge tube, and adding the purified cysteine¹⁸F-FVSB5mCi (98% methanol) 100. mu.L and sodium borate buffer (PH8.5) 100. mu.L, after mixing, react for 30min at 35 ℃. After completion of the reaction, 200. mu.L of 5% acetic acid was added to the crude reaction system for dilution, followed by liquid phase separation. The liquid chromatography conditions were the same as in example 1.

¹⁸HPLC charts of F-FVSB- (L) cysteine are shown in FIG. 3 and FIG. 4, in which the detector of FIG. 3 is a radioactivity detector, the detector of FIG. 4 is an ultraviolet detector, and the detection wavelength is 254 nm.

Collecting eluate with peak time of 7.8min, diluting the collected eluate with water until ACN content is below 5%, hanging on HLB solid phase extraction column, washing with 10mL water, eluting with 200 μ L ethanol, and collecting eluate to obtain final product¹⁸F-FVSB-L-cysteine.

Obtained in this example¹⁸F-FVSB-L-cysteine M/z [ M + H ]]+Calcd for C₁₁H₁₅O₄FNS₂308.04，find 308.05。

Example 3

This example discloses compounds of formula I-2¹⁸The preparation of F-FVSB-GSH has the reaction formula:

placing GSH 1mg into 2mL centrifuge tube, adding purified GSH¹⁸F-FVSB5mCi (98% methanol) 100. mu.L and sodium borate buffer (pH8.5) 100. mu.L, after mixing, react at 35 ℃ for 30 min. After the reaction, 200. mu.L of 5% acetic acid was added to the crude reaction system to dilute the mixture, followed by liquid phase separation to prepare a preparation. The liquid chromatography conditions were the same as in example 1.

¹⁸The HPLC chart of F-FVSB-GSH is shown in FIG. 5 and FIG. 6, wherein the detector of FIG. 5 is a radioactivity detector, the detector of FIG. 6 is an ultraviolet detector, and the detection wavelength is 254 nm.

Collecting eluate with peak time of 7.7min, diluting the collected eluate with water until ACN content is below 5%, hanging on HLB solid phase extraction column, washing HLB column with 10mL water, eluting with 200 μ L ethanol, and collecting eluate to obtain¹⁸F-FVSB-GSH。

Obtained in this example¹⁸F-FVSB-GSH m/z:[M+H]+Calcd for C₁₈H₂₅O₈FN₃S₂494.10，find 494.07。

Example 4

This example discloses the Compound of formula I-1 prepared in example 2¹⁸F]The imaging experiment of-FVSB- (L) cysteine specifically includes: human non-small cell lung adenocarcinoma cell line H1975 5% CO at 37 ℃₂Incubations were performed in an incubator and grown in PRMI 1640 medium supplemented with 10% fetal bovine serum and 1% double antibody. The right side of the female athymic nude mouse is inoculated with 2X 10 mixed with PRMI 1640 culture medium with the total volume of 100 mu L⁶H1975 cells, allowing tumor to develop for 1-2 weeks until the size reaches 200-³This is approximately 2-3 weeks after inoculation.

The dose of the tail vein injection of the female athymic nude mouse is about 80-150uCi, and after 0.5h of vein injection, the female athymic nude mouse is anesthetized by 2 percent isoflurane gas and is subjected to PET-CT static scanning imaging for 20 min.

The results are shown in FIG. 7, from which it can be seen that:¹⁸after F-FVSB- (L) cysteine is injected into the tail vein of a nude mouse for 0.5h, through the static scanning of a small animal PET-CT, the medicine is mainly metabolized from the kidney, the metabolic speed is high, the maximum tumor uptake value is 1 +/-0.0048% ID/g, the maximum muscle uptake value is 0.02 +/-0.0018% ID/g, the liver uptake value is low, the muscle/tumor uptake ratio is 50, the image background is low, the resolution is high, and the target tissue is clearly developed.

Example 5

This example discloses the use of the compound of formula I-2 prepared in example 3¹⁸The imaging experiment of the F-FVSB-GSH specifically comprises the following steps:

human non-small cell lung adenocarcinoma cell line H1975 5% CO at 37 ℃₂Incubations were performed in an incubator and grown in PRMI 1640 medium supplemented with 10% fetal bovine serum and 1% double antibody. The right side of the female athymic nude mouse is inoculated with 100 mu L of 2X 10 mixed with PRMI 1640 medium (1:1 volume ratio)⁶H1975 cells, allowing tumor to develop for 1-2 weeks until the size reaches 200-³This is approximately 2-3 weeks after inoculation.

The dose of the tail vein injection of the female athymic nude mouse is about 80-150uCi, after half an hour of vein injection, the female athymic nude mouse is anesthetized by 2 percent isoflurane gas and is subjected to PET-CT static scanning imaging for 20 min.

The results are shown in FIG. 8, from which it can be seen that:¹⁸after the F-FVSB-GSH is injected into the tail vein of a nude mouse for 0.5h, the static scanning of a small animal PET-CT shows that the medicine is mainly metabolized from the kidney, the metabolic speed is high, the maximum uptake value of tumor is 1.5 +/-0.0077% ID/g, the maximum uptake value of muscle is 0.4 +/-0.02% ID/g, the liver uptake value is low, the muscle/tumor uptake ratio is 3.75, the image background is low, the resolution is high, and the target tissue is clearly developed.

Finally, it should be noted that: the above embodiments are only preferred embodiments of the present invention to illustrate the technical solutions of the present invention, but not to limit the technical solutions, and certainly not to limit the patent scope of the present invention; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention; that is, the technical problems to be solved by the present invention, which are not substantially changed or supplemented by the spirit and the concept of the main body of the present invention, are still consistent with the present invention and shall be included in the scope of the present invention; in addition, the technical scheme of the invention is directly or indirectly applied to other related technical fields, and the technical scheme is included in the patent protection scope of the invention.