Sulfonyl fluoro-compound and application thereof
阅读说明：本技术 一种磺酰氟基化合物及其应用 (Sulfonyl fluoro-compound and application thereof ) 是由 吴泽辉 吉训明 孙雨丽 陈华龙 程雪波 蒋增 郑伟 杨庭钰 于子越 于 2021-07-14 设计创作，主要内容包括：本发明公开了一种磺酰氟基化合物,本发明设计并合成的一系列磺酰氟基化合物,与已报道的靶向氨基酸转运载体、成纤维细胞活化蛋白、囊泡单胺转运蛋白2、五羟色胺受体、前列腺特异性膜抗原蛋白变化相关疾病的PET显像剂相比,具有标记条件简单,产率高,亲和性高的优势。本发明属于放射性药物化学与核医学技术领域。(Compared with the reported PET imaging agents for diseases related to changes of targeted amino acid transport carriers, fibroblast activation proteins, vesicular monoamine transporter 2, serotonin receptors and prostate specific membrane antigen proteins, the series of sulfonyl fluoride compounds designed and synthesized by the invention have the advantages of simple labeling conditions, high yield and high affinity. The invention belongs to the technical field of radiopharmaceutical chemistry and nuclear medicine.)
1. A sulfonyl fluoride-based compound having a structure represented by the following formula 1 to formula 5:
in the formula 1, R1Is H atom or methyl;
R2is methylene, alkyl having 1-10 carbon atoms, alkoxy having 1-10 carbon atoms, branched alkylamide having 1-10 carbon atoms, guanidine having 1-10 carbon atoms, or R2Has the advantages of The structure shown;
in the formula 4, R3Is CH2Or CF2；
R4Has the advantages ofThe structure shown in the specification, wherein R5、R6、R7Each independently selected from alkyl or alkoxy groups having 1 to 10 carbon atoms;
in the formula 5, R8Selected from alkyl, alkoxy, amido and ester groups with 0-10 carbon atoms.
2. The compound of claim 1, wherein the fluorine atom in the sulfonyl fluoride group in formulas 1 to 5 is selected from the group consisting of19F or18F, preferably18F。
3. The compound of claim 1, wherein the compound of formula 1 has a structure represented by formula a-formula d:
4. the compound of claim 1, wherein the compound of formula 4 has the structure of formula e, formula j:
5. the compound of claim 1, wherein the compound of formula 5 has the structure of formula k-formula m:
6. use of a compound according to claims 1-5 for the preparation of a medicament, medicament or kit for the diagnosis of tumours and neurological related diseases.
7. The use of claim 6, wherein the medicament, medicament or kit comprises a positron emission tomography imaging technique imaging agent.
8. The use of claim 7, wherein the imaging agent comprises18F-labeled compounds represented by formula 1 to formula 5.
9. The use according to claim 6, wherein the tumor and nervous system related diseases comprise one or more of Parkinson's disease, brain glioma, prostate cancer and breast cancer.
Technical Field
The invention belongs to the technical field of radiopharmaceutical chemistry and nuclear medicine, and particularly relates to a sulfonyl fluoride compound and application thereof.
Background
Positron Emission Tomography (PET) technology can provide molecular level and physiological information of different canceration parts from the outside of a body in a noninvasive, quantitative and dynamic mode, has high sensitivity, strong specificity and good safety, and can be used for whole-body imaging, and becomes a powerful means for clinical diagnosis of tumors, cardiovascular and cerebrovascular diseases and the like at present. Two important conditions for PET imaging are achieved: PET instruments and molecular probes. The study of specific imaging probes is currently a major and difficult point. The method is an important way for solving the early diagnosis of related diseases by searching effective targets related to diseases, designing molecules specifically bound with the targets, developing an ideal radioactive molecular imaging probe and developing a novel PET imaging medicine.
With the sustainable development of economy in China, PET imaging instruments are greatly increased, the domestic PET instruments are put on the market recently, a rapid development situation is formed, the demand on novel PET molecular probes is particularly remarkable, and the bottleneck effect is more prominent. The research and development of a novel PET molecular probe becomes a preoccupation, and the research, fusion and development of the modern radiochemical rapid synthesis technology, the radiopharmaceutical analysis technology, the disease animal model and the PET imaging technology matched with the PET molecular probe can ensure that the clinical transformation of the PET molecular probe is smoothly and effectively carried out. Among the radionuclides, those that do not have,18long half-life period of F, low electron energy, high resolution of PET image, and18f atom is small and not shadowIn response to the nature of the originally targeted molecule, and therefore,18f-labelled drugs are widely developed.18The F-labeled radioactive molecular probe is usually an activated ester nucleophilic heating reaction which needs a high energy barrier, and the labeling conditions are usually harsh, so that the F-labeled radioactive molecular probe is not favorable for clinical transformation. The urgent need to develop18The PET new drug with simple labeling condition F meets the requirements of people health and clinical transformation.
Disclosure of Invention
The invention solves the technical problem of providing a high-affinity sulfonyl fluoride-based compound and application thereof, and the compound is used for a PET developer and has the remarkable advantages of simple marking condition, high yield and high affinity.
In order to achieve the above object, the first aspect of the present invention provides the following solutions:
a sulfonyl fluoride-based compound having a structure represented by the following formula 1 to formula 5:
in the formula 1, R1Is H atom or methyl;
R2is methylene, alkyl having 1-10 carbon atoms, alkoxy having 1-10 carbon atoms, branched alkylamide having 1-10 carbon atoms, guanidine having 1-10 carbon atoms, or R2Has the advantages of The structure shown;
in the formula 4, R3Is CH2Or CF2；
R4Has the advantages ofThe structure shown in the specification, wherein R5、R6、R7Each independently selected from alkyl or alkoxy groups having 1 to 10 carbon atoms;
in the formula 5, R8Selected from alkyl, alkoxy, amido and ester groups with 0-10 carbon atoms.
As an example of the present invention, the fluorine atom in the sulfonyl fluoride group in the formula 1 to formula 5 is selected from19F or18F, preferably18F。
As an example of the present invention, the compound having formula 1 has a structure represented by the following formula a to formula d:
as an example of the present invention, the compound of formula 4 has a structure represented by formula e below, formula j below:
as an example of the present invention, the compound of formula 5 has a structure represented by the following formula k-formula m:
in a second aspect, the invention provides the use of a compound as described above in the manufacture of a medicament, medicament or kit for the diagnosis of tumours and neurological related disorders.
As an example of the invention, the medicament, medicament or kit comprises a positron emission tomography imaging technique imaging agent.
As an example of the present invention, the imaging agent comprises18F-labeled compounds represented by formula 1 to formula 5.
As an example of the present invention, the tumor and nervous system related diseases include one or more of Parkinson's disease, brain glioma, prostate cancer and breast cancer.
The technical scheme provided by the invention at least brings the following beneficial effects:
compared with the reported PET imaging agents for diseases related to changes of targeted amino acid transport carriers, fibroblast activation proteins, vesicular monoamine transporter 2, serotonin receptors and prostate specific membrane antigen proteins, the series of sulfonyl fluoride-based compounds designed and synthesized by the invention have the advantages of simple labeling conditions, high yield and high affinity.
Drawings
FIG. 1 is an image showing the result of 30min after injecting the sulfonyl fluoride-based compound prepared in example 1 into a tumor-bearing mouse in example 26 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in further detail below.
The "purification" in the present invention is intended to remove18Excess by-product of F-fluoride, and concentration and capture of the reaction product. Purification is carried out by any method known to the person skilled in the art suitable for radiotracers, such as chromatography, HPLC, solid phase extraction cartridge or solid phase extraction column.
The preparation method of the sulfonyl fluoride-based compound comprises the following steps:
example 1(S) -2-amino-3- (4- ((fluorosulfonyl) oxy) phenyl) propionic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) synthesis of (S) -2- (((tert-butyloxycarbonyl) amino) tert-butyl-3- (4- ((fluorosulfonyl) oxy) phenyl) propionate
The compound (S) -2-amino-3- (4- ((fluorosulfonyl) oxy) phenyl) propanoic acid (0.5g,1.48mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.59mL,5.93mmol) was added, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (0.36g, 58.4%). 1H NMR (300MHz, CDCl3) δ 7.30(s,4H),5.09(d, J ═ 4.9Hz,1H),4.47(d, J ═ 5.5Hz,1H),3.09(s,2H),1.42(d, J ═ 7.2Hz,18H), 13C NMR (75MHz, CDCl3) δ 170.46,155.03,148.97,137.66,131.43,120.67,82.52,79.72,77.44,77.02,76.59,54.67,38.13,28.25,27.89 HRMS calcd for C18H26FNO7S 419.1414; found,420.1421[ M + H [ ]]+.
(2) Synthesis of (S) -2-amino-3- (4- ((fluorosulfonyl) oxy) phenyl) propanoic acid
The compound ((S) -2- (((tert-butoxycarbonyl) amino) tert-butyl-3- (4- ((fluorosulfonyl) oxy) phenyl) propionate (0.5g,1.19mmol) was added to 2mL of TFA, reacted at room temperature for 3H, followed by removal of TFA and HPLC semipreparative isolation to give the product (0.28g, 89.1%). sup.1H NMR (300MHz, MeOD) delta 7.52(d, J ═ 8.3Hz,2H),7.42(d, J ═ 7.9Hz,2H),5.45(S,1H),4.29(S,1H),3.32(M,1H).13C NMR (75MHz, MeOD) delta 169.95,167.86,149.62,135.82,131.52,121.08,118.89,114.53,53.67,48.52,48.24,47.96,47.67,47.39,47.10,46.82,35.17,26.63.HRMS cad for, C9H 10O 5, 5S 263.0264 6 [ 264.1257 + 67M + 264.1257H ] F H]+.
Example 2(2S, 4S) -2, 5-diamino-4- (4- ((fluorosulfonyl) oxy) benzyl) -5-oxopentanoic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) synthesis of tert-butyl (2S, 4S) -2- (((tert-butoxycarbonyl) amino) -4- (4- ((fluorosulfonyl) oxy) benzyl) -5-oxo-5- ((2,4, 5-trimethoxybenzyl) amino) pentanoate
The compound tert-butyl (2S, 4S) -2- (((tert-butoxycarbonyl) amino) -4- (4-hydroxybenzyl) -5-oxo-5- ((2,4, 5-trimethoxybenzyl) amino) pentanoate (0.2g,0.34mmol) was dissolved in 20mL of dichloromethane, placed in ice water, followed by addition of triethylamine (0.13mL,1.3mmol), and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (0.13g, 56.2%).1H NMR(300MHz,CDCl3)δ7.09(d,J＝6.1Hz,2H),6.92(d,J＝6.6Hz,2H),5.94(s,2H),5.19(s,1H),4.44(s,1H),4.13(s,1H),3.97(m,2H),3.63(m,9H),2.77(s,2H),2.23(s,2H),1.89(d,J＝2.6Hz,2H),1.32(s,18H).13C NMR(75MHz,CDCl3)δ172.63,171.51,170.90,160.77,158.97,155.55,148.31,140.50,130.77,130.65,120.24,120.11,106.27,90.30,81.77,79.45,77.67,77.25,76.82,60.16,55.45,55.01,52.52,46.30,37.22,35.27,31.95,28.11,27.72,20.75,13.99.HRMS calcd for；C31H43FN2O11S670.2572found,671.2674[M+H]+.
(2) Synthesis of (2S, 4S) -2, 5-diamino-4- (4- ((fluorosulfonyl) oxy) benzyl) -5-oxopentanoic acid
The compound tert-butyl (2S, 4S) -2- (((tert-butoxycarbonyl) amino) -4- (4- ((fluorosulfonyl) oxy) benzyl) -5-oxo-5- ((2,4, 5-trimethoxybenzyl) amino) pentanoate (0.2g,0.3mmol) was added to 2mL TFA, reacted at room temperature for 3H, followed by TFA removal, HPLC semipreparative isolation (52mg, 51.4%). 1H NMR (300MHz, DMSO) δ 8.03(S,1H),7.47(d, J ═ 8.2Hz,2H),7.38(d, J ═ 8.4Hz,2H),6.87(S,1H),3.75(S,2H),2.90(d, J ═ 12.4Hz,3H),2.64(d, J ═ 8.0Hz,1H),1.68(S,2H), ms, 3515H, FN 15, un, 2H,335.1623[M+H]+.
example 32-amino-3- (4- ((fluorosulfonyl) oxy) phenyl) -2-methylpropanoic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) synthesis of tert-butyl 2- ((tert-butoxycarbonyl) amino) -3- (4- ((fluorosulfonyl) oxy) phenyl) -2-methylpropionate
The compound tert-butyl 2- ((tert-butoxycarbonyl) amino) -3- (4-hydroxyphenyl) -2-methylpropionate (0.2g, 0.56mmol) was dissolved in 20mL of dichloromethane, placed in ice water, and triethylamine (2.2mL, 2.27mmol) was added thereto, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. Then the reaction solution was added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain a product (0.15g, 63.4%). HRMS calcd for; C19H28FNO7S 433.1571found,434.1689[ M + H ]]+.
(2) Synthesis of 2-amino-3- (4- ((fluorosulfonyl) oxy) phenyl) -2-methylpropanoic acid
The compound tert-butyl 2- ((tert-butoxycarbonyl) amino) -3- (4- ((fluorosulfonyl) oxy) phenyl) -2-methylpropionate (0.1g,0.23mmol) was added to 2mL of TFA and reacted at room temperature for 3h before removing TFA and separation by HPLC semipreparative (39mg, 62.4%). HRMS calcd for; C10H12FNO5S277.0420found,278.1231[ M + H ]]+。
Example 4(2S, 4S) -2-amino-4- (4- ((fluorosulfonyl) oxy) benzyl) -5-guanidinopentanoic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) synthesis of tert-butyl (2S, 4S) -5- ((E) -1, 2-bis (tert-butoxycarbonyl) -3- (4-methoxybenzyl) guanidino) -2- ((tert-butoxycarbonyl) amino) -4- (4- ((fluorosulfonyl) oxy) benzyl) pentanoate
The compound tert-butyl (2S, 4S) -5- ((E) -1, 2-bis (tert-butoxycarbonyl) -3- (4-methoxybenzyl) guanidino) -2- ((tert-butoxycarbonyl) amino) -4- (4-hydroxybenzyl) pentanoate (0.2g, 0.26mmol) was dissolved in 20mL of dichloromethane, placed in ice water, and triethylamine (0.1mL, 1.05mmol) was added thereto, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (0.11g, 53.8%).1H NMR(300MHz,CDCl3)δ7.35–7.19(m,6H),6.89(d,J＝8.6Hz,2H),5.09(s,1H),4.34(s,2H),3.80(s,3H),3.67(s,2H),2.96(d,J＝11.6Hz,1H),2.53(m,1H),2.16(s,1H),1.43(dd,J＝18.0,14.6Hz,37H).13C NMR(75MHz,CDCl3)δ171.89,159.42,155.75,153.44,148.47,140.62,131.28,129.08,120.52,114.36,82.84,82.03,79.76,77.47,77.25,77.05,76.62,55.25,51.72,50.58,47.37,36.86,36.61,35.98,35.39,28.32,28.17,27.99,27.92,26.39.HRMS calcd for；C40H59FN4O12S 838.3834found,839.4521[M+H]+.
(2) Synthesis of (2S, 4S) -2-amino-4- (4- ((fluorosulfonyl) oxy) benzyl) -5-guanidinopentanoic acid
The compound tert-butyl (2S, 4S) -5- ((E) -1, 2-bis (tert-butoxycarbonyl) -3- (4-methoxybenzyl) guanidino) -2- ((tert-butoxycarbonyl) amino) -4- (4- ((fluorosulfonyl) oxy) benzyl) pentanoate (0.1g,0.12mmol) was added to 2mL TFA and reacted at room temperature for 3h before removing TFA and isolating by HPLC semi-preparative (24mg, 56.3%).1H NMR(300MHz,D2O)δ7.71(d,J＝8.1Hz,2H),6.99(d,J＝8.1Hz,2H),3.82(t,J＝7.2Hz,1H),3.12-2.90(m,2H),2.60(t,J＝6.9Hz,2H),2.27–2.05(m,1H),1.91–1.55(m,2H).HRMS calcd for；C13H19FN4O5S 362.1060found,363.1123[M+H]+.
Example 54- ((2-aminophenyl) thio) -3- ((dimethylamino) methyl) phenylthiofluoroacid ester
The structural formula is as follows:
the synthetic route is as follows:
the compound 4- ((2-aminophenyl) thio) -3- ((dimethylamino) methyl) phenol (0.1g,0.36mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.15mL,1.45mmol) was added, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain a product (84mg, 64.2%). 1H-NMR (300MHz, CDCl3): δ 7.35(dd, J ═ 7.6,1.5Hz,1H),7.14(dd, J ═ 7.6,1.5Hz,1H),6.86(d, J ═ 8.4Hz,1H), 6.74-6.67 (m,3H),6.56-6.51(m,1H),4.51(s,3H),3.54(s,2H),2.21(s,6H), HRMS calcd for; C15H17FN2O3S2356.0665found, 357.1589[ M + H]+.
EXAMPLE 6(2R, 3R, 11bR) -2-hydroxy-3-isobutyl-10-methoxy-11 b-methyl-1, 3,4,6,7,11 b-hexahydro-2H-pyridinyl [2,1-a ] isoquinolin-9-yl-thiofluoride
The structural formula is as follows:
the synthetic route is as follows:
(1) synthesis of (2R, 3R, 11bR) -3-isobutyl-10-methoxy-11 b-methyl-2- ((tetrahydro-2H-pyran-2-yl) oxy) -1,3,4,6,7,11 b-hexahydro-2H-pyrido [2,1-a ] isoquinolin-9-yl-thiofluo-ride
The compound (2R, 3R, 11bR) -3-isobutyl-10-methoxy-11 b-methyl-2- ((tetrahydro-2H-pyran-2-yl) oxy) -1,3,4,6,7,11 b-hexahydro-2H-pyrido [2, 1-a)]Isoquinolin-9-ol (0.2g,0.49mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.2mL,1.98mmol) was added and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (0.15g, 65.7%). HRMS calcd for; C24H36FNO6S 485.2247found,486.2347[ M + H ]]+.
(2) Synthesis of (2R, 3R, 11bR) -2-hydroxy-3-isobutyl-10-methoxy-11 b-methyl-1, 3,4,6,7,11 b-hexahydro-2H-pyridinyl [2,1-a ] isoquinolin-9-yl-thiofluoride
The compound (2R, 3R, 11bR) -3-isobutyl-10-methoxy-11 b-methyl-2- ((tetrahydro-2H-pyran-2-yl) oxy) -1,3,4,6,7,11 b-hexahydro-2H-pyrido [2, 1-a)]Isoquinolin-9-yl thiofluoroacid ester (0.1g,0.2mmol) was dissolved in 10mL of ethanol, and PPTS (10mg, 0.02mmol) was added at 50 ℃ for 3h, followed by column chromatography to give the product (63mg, 76.8%).1HNMR(400MHz,CDCl3)δ6.67(s,1H),6.65(s,1H),3.89(s,3H),3.44-3.35(m,1H),3.17-2.96(m,4H),2.65-2.56(m,2H),2.49-2.43(m,1H),2.01(t,J＝2.01Hz,1H),1.79-1.69(m,2H),1.56-1.45(m,3H),1.12-1.09(m,1H),0.98-0.92(m,6H).HRMS calcd for；C19H28FNO5S 401.1672found,402.2675[M+H]+.
Example 7(S) -4- ((2- (2-cyano-4, 4-difluoropyrrolidin-1-yl) -2-oxoethyl) carbamoyl) quinolin-6-yl thiofluoroacid ester
The structural formula is as follows:
the synthetic route is as follows:
will be transformed intoThe compound (S) -N- (2- (2-cyano-4, 4-difluoropyrrolidin-1-yl) -2-oxoethyl) -6-hydroxyquinoline-4-carboxamide (0.2g, 0.55mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.2g, 2.22mmol) was added, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (0.16g, 69.5%). HRMS calcd for; C17H13F3N4O5S 442.0559found,443.1562[ M + H ]]+.
Example 8(S) -4- ((2- (2-Cyanopyrrolidin-1-yl) -2-oxoethyl) carbamoyl) quinolin-6-yl-thiofluoroacid ester
The structural formula is as follows:
the synthetic route is as follows:
the compound (S) -N- (2- (2-cyanopyrrolidin-1-yl) -2-oxoethyl) -6-hydroxyquinoline-4-carboxamide (0.1g,0.3 mmol) was dissolved in 20mL of dichloromethane, placed in ice water and triethylamine (0.1mL, 1.23mmol) was added and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (80mg, 65.8%). HRMS calcd for; C17H15FN4O5S 406.0747found,407.1249[ M + H ]]+.
Example 9(S) -4- (4- (3- ((4- ((2- (2-cyano-4, 4-difluoropyrrolidin-1-yl) -2-oxoethyl) carbamoyl) quinolin-6-yl) oxy) propyl) piperazine-1-carbonyl) phenylthiofluoroacid ester
The structural formula is as follows:
the synthetic route is as follows:
the compound (S) -N- (2- (2-cyano-4, 4-difluoropyrrolidin-1-yl) -2-oxoethyl) -6- (3- (4- (4-hydroxybenzoyl) piperazin-1-yl) propoxyquinoline-4-carboxamide (0.1g,0.16mmol) was dissolved in 20mL of dichloromethane, placed in ice-water, triethylamine (0.2mL,0.64mmol) was added, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain a product (51mg, 46.7%). HRMS calcd for; C31H31F3N6O7S 688.1927found,689.2043[ M + H ]]+.
Example 10(S) -4- (2- (4- (3- ((4- ((2- (2-cyanopyrrolidine) -1-yl) -2-oxyethyl) carbamoyl) quinolin-6-yl) oxy) propyl) piperazin-1-yl) -2-oxyethyl) phenylthiofluoride salt
Example 10 was obtained with reference to the synthetic routes of examples 8 and 9, with an overall yield of 28%. HRMS calcd for; C32H35FN6O7S 666.2272found,667.2353[ M + H ]]+.
Example 11(S) -4- (2- (4- (3- ((4- ((2- (2-cyano-4, 4-difluoropyrrolidin-1-yl) -2-oxyethyl) carbamoyl) quinolin-6-yl) oxy) propyl) piperazin-1-yl) -2-oxyethyl) phenylthiofluoride salt
Example 11 was obtained with reference to the synthetic routes of examples 8 and 9, with an overall yield of 23%. HRMS calcd for; C32H33F3N6O7S 702.2084found,703.3541[ M + H ]]+.
Example 12(S) -4- (4- (3- ((4- ((2- (2-cyanopyrrolidin-1-yl) -2-oxoethyl) carbamoyl) quinolin-6-yl) oxy) propyl) piperazine-1-carbonyl) phenylthiofluoride
The structural formula is as follows:
the synthetic route is as follows:
the compound (S) -N- (2- (2-cyanopyrrolidin-1-yl) -2-oxyethyl) -6- (3- (4- (4-hydroxybenzoyl) piperazin-1-yl) propoxy) quinoline-4-carboxamide (0.1g,0.17mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.7g,0.7mmol) was added, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (53mg, 49.1%). HRMS calcd for; C31H33FN6O652.2115found, 635.2241[ M + H]+.
Example 13(((S) -1-carboxy-5- (4- ((fluorosulfonyl) oxy) benzamido) pentyl) carbamoyl) -L-glutamic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) synthesis of di-tert-butyl (((S) -1- (tert-butoxy) -6- (4-hydroxybenzamido) -1-oxohexyl-2-yl) carbamoyl) -L-glutamic acid
Di-tert-butyl (((S) -6-amino-1- (tert-butoxy) -1-oxyhexyl-2-yl) carbamoyl) -L-glutamic acid (0.5g, 1mmol) was dissolved in 20mL of dichloromethane, and then 1mL of triethylamine and p-hydroxybenzoic acid (0.14g,1mmol), 10mg of HOBt and EDCI (0.38g,2mmol) were added, followed by reaction overnight.The reaction solution was poured into 10mL of water, and the resulting solution was separated, dried, and subjected to column chromatography to obtain the product (0.32g, 53.7%). 1H NMR (300MHz, CDCl3) δ 7.69(d, J ═ 7.8Hz,2H),7.12(s,1H),6.87(d, J ═ 7.8Hz,2H),5.55(s,2H),4.28(s,2H),3.37(s,2H),2.33(s,2H),1.64(m,18H), 13C NMR (75MHz, CDCl3) δ 172.51,168.09,159.96,157.41,129.16,115.45,82.32,81.89,80.78,77.42,77.20,77.00,76.57,53.46,53.16,39.85,32.60,31.62,28.89,28.07,27.99,27.96,22.73.HRMS calcd for; C31H49N3O9607.3469found, 608.4519[ M + H]+.
(2) Synthesis of di-tert-butyl (((S) -1- (tert-butoxy) -6- (4- ((fluorosulfonyl) oxy) benzamido) -1-oxyhexyl-2-yl) carbamoyl) -L-glutamic acid
The compound di-tert-butyl (((S) -1- (tert-butoxy) -6- (4-hydroxybenzamido) -1-oxyhexyl-2-yl) carbamoyl) -L-glutamic acid (0.2g, 0.33mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.12g, 1.3mmol) was added and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. Then the reaction solution was added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain a product (0.12g, 54.2%).1H NMR(300MHz,CDCl3)δ8.08(d,J＝8.8Hz,3H),7.35(d,J＝8.5Hz,2H),6.12(s,1H),5.64(s,1H),4.05(m,2H),3.30(s,2H),2.20(s,2H),1.50(m,18H).13C NMR(75MHz,CDCl3)δ174.24,172.27,171.86,165.79,157.70,151.57,135.01,130.09,120.60,82.65,81.35,80.62,77.56,77.13,76.71,60.31,54.01,52.99,39.99,32.16,31.36,29.07,27.91,27.85,27.74,27.63,23.87,20.92,14.08.HRMS calcd for；C31H48FN3O11S 689.2994found,690.1259[M+H]+.
(3) Synthesis Di-tert-butyl (((S) -1- (tert-butoxy) -6- (4- ((fluorosulfonyl) oxy) benzamido) -1-oxyhexyl-2-yl) carbamoyl) -L-glutamic acid (0.1g,0.14mmol) was added to 2mL of TFA and reacted at room temperature for 3h, after which the TFA was removed and isolated by HPLC semipreparative (51mg, 68.2%).1H NMR(300MHz,MeOD)δ8.00(d,J＝8.8Hz,2H),7.56(d,J＝8.5Hz,2H),4.31(d,J＝5.1Hz,2H),3.41(s,2H),2.42(t,J＝7.0Hz,2H),2.19–2.12(m,1H),1.89-1.85(m,2H),1.70-1.65(m,4H),1.54-1.50(m,2H),1.32-1.26(m,1H).13C NMR(75MHz,MeOD)δ175.16,167.01,158.75,151.84,135.20,129.53,120.85,52.60,52.15,48.46,48.18,47.89,47.61,47.33,47.04,46.76,39.50,31.79,29.70,28.47,27.48,22.61.HRMS calcd for；C19H24FN3O11S 521.1116found,522.1124[M+H]+.
Example 14(((S) -1-carboxy-5- (2- (4- ((fluorosulfonyl) oxy) phenyl) acetamido) pentyl) carbamoyl) -L-glutamic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) di-tert-butyl (((S) -1- (tert-butoxy) -6- (2- (4-hydroxyphenyl) acetamido) -1-oxohexyl-2-yl) carbamoyl) -L-glutamic acid
Di-tert-butyl (((S) -6-amino-1- (tert-butoxy) -1-oxyhexyl-2-yl) carbamoyl) -L-glutamic acid (0.5g, 1mmol) was dissolved in 20mL of dichloromethane, and then 1mL of triethylamine and p-hydroxyphenylacetic acid (0.15g,1mmol), 10mg of HOBt and EDCI (0.38g,2mmol) were added, followed by overnight reaction. The reaction solution was poured into 10mL of water, and the resulting solution was separated, dried, and subjected to column chromatography to obtain the product (0.31g, 49.2%).1H NMR(300MHz,CDCl3)δ7.25-7.15(m,1H),7.05–6.95(m,2H),6.85–6.77(m,2H),6.27(s,1H),5.64(d,J＝40.9Hz,2H),4.34(s,1H),4.22(s,1H),3.52-3.42(m,2H),3.16(d,J＝6.4Hz,2H),2.31(s,2H),1.90–1.78(m,2H),1.67–1.58(m,2H),1.52-1.44(m,30H).13C NMR(75MHz,CDCl3)δ172.79,172.68,172.46,157.48,156.10,130.40,130.28,125.85,121.74,116.01,115.82,82.30,81.68,80.68,77.49,77.06,76.64,53.53,53.04,42.62,32.28,31.62,28.75,28.20,28.04,27.97,22.45.
(2) Di-tert-butyl (((S) -1- (tert-butoxy) -6- (2- (4- ((fluorosulfonyl) oxy) phenyl) acetamide) -1-oxohexyl-2-yl) carbamoyl) -L-glutamic acid
Reacting a compound di-tert-butyl (((S)-1- (tert-butoxy) -6- (2- (4-hydroxyphenyl) acetamido) -1-oxohexyl-2-yl) carbamoyl) -L-glutamic acid (0.2g, 0.32mmol) was dissolved in 20mL of dichloromethane, placed in ice water, followed by addition of triethylamine (0.12g, 1.3mmol) and SO2A balloon of ClF gas was inserted under the liquid and reacted overnight. Then the reaction solution was added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain a product (0.1g, 48.3%).1H NMR(300MHz,CDCl3)δ7.46(d,J＝8.6Hz,2H),7.27(d,J＝7.1Hz,2H),5.97(s,1H),5.57(s,1H),4.35-4.29(m,1H),4.16-4.05(m,1H),3.66–3.54(m,2H),3.30–3.13(m,2H),2.37–2.31(m,2H),2.10(m,1H),1.88–1.68(m,2H),1.47–1.25(m,30H).13C NMR(75MHz,CDCl3)δ173.88,172.25,172.02,170.37,157.60,148.92,136.76,131.23,120.82,82.79,81.61,80.73,77.47,77.05,76.62,53.79,53.17,42.42,39.28,32.36,31.54,28.84,27.99,27.94.
(3) ((S) -1-carboxy-5- (2- (4- ((fluorosulfonyl) oxy) phenyl) acetamido) pentyl) carbamoyl) -L-glutamic acid
Di-tert-butyl (((S) -1- (tert-butoxy) -6- (2- (4- ((fluorosulfonyl) oxy) phenyl) acetamide) -1-oxohexyl-2-yl) carbamoyl) -L-glutamic acid (0.1g,0.14mmol) was added to 2mL of TFA and reacted at room temperature for 3h before removing TFA and isolating by HPLC semipreparative (42mg, 55.3%).1H NMR(300MHz,MeOD)δ7.48(d,J＝8.8Hz,2H),7.40(d,J＝8.7Hz,2H),4.36–4.27(m,2H),3.58(s,2H),3.21(t,J＝6.7Hz,2H),2.46–2.39(m,2H),2.16–2.05(m,1H),1.89–1.82(m,1H),1.55(m,2H).13C NMR(75MHz,MeOD)δ175.07,174.50,171.67,158.72,149.12,136.98,130.91,120.58,99.99,52.56,52.15,48.44,48.16,48.09,48.06,48.03,48.00,47.97,47.94,47.87,47.79,47.75,47.59,47.52,47.49,47.46,47.43,47.40,47.37,47.31,47.22,47.19,47.16,47.02,46.92,46.89,46.86,46.84,46.80,46.74,41.53,38.97,31.77,29.72,28.41,27.50,22.53.
Example 15(15S,19S) -1- (4- ((fluorosulfonyl) oxy) phenyl) -2,9, 17-trioxa-3, 10,16, 18-tetraazaeicosane-15, 19, 21-tricarboxylic acid
The structural formula is as follows:
the synthetic route is as follows:
(1) tri-tert-butyl (15S,19S) -1- (4-hydroxyphenyl) -2,9, 17-trioxa-3, 10,16, 18-tetraazadocosane-15, 19, 21-tricarboxylate
Di-tert-butyl (((S) -6- (6-aminocaproamide) -1- (tert-butoxy) -1-oxohexyl-2-yl) carbamoyl) -L-glutamic acid (0.4g, 0.67mmol) was dissolved in 20mL of dichloromethane, and then 1mL of triethylamine and p-hydroxyphenylacetic acid (0.1g,0.67mmol), 10mg of HOBt and EDCI (0.38g,2mmol) were added, followed by overnight reaction. The reaction solution was poured into 10mL of water, and the product was obtained by separation, drying and column chromatography (0.25g, 52.2%).1H NMR(300MHz,CDCl3)δ7.05(d,J＝8.2Hz,2H),6.82(d,J＝8.2Hz,2H),6.69(s,1H),6.13(s,1H),5.91–5.86(m,2H),4.32–4.28(m,2H),3.46(s,2H),3.21–3.11(m,4H),2.32(s,2H),2.10–1.98(m,4H),1.82–1.76(m,2H),1.57-1.34(m,39H).13C NMR(75MHz,CDCl3)δ173.83,172.80,172.53,172.48,172.39,157.67,156.36,130.39,125.75,116.03,82.08,81.62,80.65,77.46,77.24,77.04,76.61,53.39,53.04,42.83,39.28,39.03,36.14,32.34,31.67,28.92,28.70,28.05,27.99,26.07,25.14,22.50.
(2) Tri-tert-butyl (15S,19S) -1- (4- ((fluorosulfonyl) oxy) phenyl) -2,9, 17-trioxa-3, 10,16, 18-tetraazaeicosane-15, 19, 21-tricarboxylate
The compound tri-tert-butyl (15S,19S) -1- (4-hydroxyphenyl) -2,9, 17-trioxa-3, 10,16, 18-tetraazadocosane-15, 19, 21-tricarboxylate (0.2g, 0.27mmol) was dissolved in 20mL of dichloromethane, placed in ice water, triethylamine (0.1g, 1mmol) was added, and SO was added2A balloon of ClF gas was inserted under the liquid and reacted overnight. The reaction solution was then added to water, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography to obtain the product (0.11g, 53.1%).1H NMR(300MHz,CDCl3)δ7.38(d,J＝8.6Hz,2H),7.21(d,J＝8.4Hz,2H),6.93(d,J＝7.5Hz,1H),6.85(s,1H),6.00(d,J＝8.0Hz,1H),5.87(d,J＝7.6Hz,1H),4.27–4.17(m,2H),3.51(s,2H),3.16–3.07(m,4H),2.25-2.01(m,4H),2.02(m,1H),1.89–1.11(m,2H),1.67–1.51(m,2H),1.39–1.19(m,39H).13C NMR(75MHz,CDCl3)δ173.57,172.97,172.43,172.21,170.36,157.64,148.87,136.63,131.39,131.15,121.53,120.82,82.00,81.35,80.53,77.59,77.16,76.74,53.40,52.92,42.47,39.42,38.90,35.96,32.28,31.56,29.57,28.86,28.74,28.13,27.97,27.92,26.24,25.08,22.61.
(3) (15S,19S) -1- (4- ((fluorosulfonyl) oxy) phenyl) -2,9, 17-trioxa-3, 10,16, 18-tetraazaeicosane-15, 19, 21-tricarboxylic acid
Tri-tert-butyl (15S,19S) -1- (4- ((fluorosulfonyl) oxy) phenyl) -2,9, 17-trioxa-3, 10,16, 18-tetraazaeicosane-15, 19, 21-tricarboxylate (0.1g,0.14mmol) was added to 2mL of TFA and reacted at room temperature for 3h before removing the TFA and isolating by HPLC semipreparative (36mg, 45.1%). HRMS calcd for; C26H37FN4O12S 648.2113found,649.2132[ M + H ]]+.
Example 16(S) -2-amino-3- (4- (((((fluoro-18F) sulfonyl) oxy) phenyl) propanoic acid
The structural formula is as follows:
the labeling route is as follows:
1. suction contains18F]H of (A) to (B)2 18The O (20mCi) solution is passed through a QMA column, and the resulting solution is subjected to column chromatography18F]F-Adsorbed on QMA column.
2. Aspirate 1mL K222(0.5mg)/K2CO3(1mg)MeCN/H20-5/2 solution through 0.5M NaHCO3An activated QMA column of [2 ], [18F-]Eluting from the column to the reaction tube. The eluate of appropriate activity was taken to dryness at 100 ℃ under a nitrogen purge.
3.1 mL of anhydrous acetonitrile was added to the reaction tube to evaporate to dryness, and the evaporation process was repeated 3 times, whereby dried [ alpha ], [ alpha ] can be obtained18F-]F-/K222/K2CO3The complex was capped and sealed and then cooled.
4. 0.1mg of the labeled precursor was dissolved in 1mL of acetonitrile, vortexed and added to the above dried and cooled complex. The reaction was carried out at room temperature for 1min, and the reaction mixture was cooled after the reaction was completed.
5. And adding 9mL of water into the cooled reaction solution for dilution, purifying the intermediate of the diluted solution by using an Oasis solid-phase extraction column, washing the Oasis solid-phase extraction column by using 10mL of water, and finally rinsing the small column by using 1mL of ethanol to obtain a reaction intermediate.
6. The ethanol solution was removed, 1mL TFA was added, 50 degrees, reacted for 5min, and the product was purified by HPLC (Phenomenex Gemini-Nx C18110A (250 x 4.6mm), 0.1% formic acid in 10% -90% acetonitrile 6/4, peak time 12.4min, purity > 95%.
Example 174- ((2-aminophenyl) thio) -3- ((dimethylamino) methyl) phenylthiofluoro ester-18F
The structural formula is as follows:
the labeling route is as follows:
1. suction contains18F]H of (A) to (B)2 18The O (20mCi) solution is passed through a QMA column, and the resulting solution is subjected to column chromatography18F]F-Adsorbed on QMA column.
2. Aspirate 1mL K222(0.5mg)/K2CO3(1mg)MeCN/H20-5/2 solution through 0.5M NaHCO3An activated QMA column of [2 ], [18F-]Eluting from the column to the reaction tube. Taking eluent with proper activity, and purging with nitrogen at 100 deg.CThe mixture was evaporated to dryness.
3.1 mL of anhydrous acetonitrile was added to the reaction tube to evaporate to dryness, and the evaporation process was repeated 3 times, whereby dried [ alpha ], [ alpha ] can be obtained18F-]F-/K222/K2CO3The complex was capped and sealed and then cooled.
4. 0.05mg of the labeled precursor was dissolved in 1mL of acetonitrile, vortexed and added to the above dried and cooled complex. The reaction was carried out at room temperature for 1min, and the reaction mixture was cooled after the reaction was completed.
5. The cooled reaction solution was diluted with 9mL of water, the diluted solution was purified by Oasis solid phase extraction column for intermediate purification and by washing with 10mL of water, and finally by rinsing the column with 1mL of ethanol and by HPLC purification (Phenomenex Gemini-Nx C18110A (250 × 4.6mm), 0.1% formic acid/acetonitrile 10% -90%: 65/35, with a peak time of 10.5min and a purity of more than 95%.
Example 18(S) -4- ((2- (2-Cyanopyrrolidin-1-yl) -2-oxoethyl) carbamoyl) quinolin-6-yl thiofluoride-18F
The structural formula is as follows:
the labeling route is as follows:
1. suction contains18F]H of (A) to (B)2 18The O (20mCi) solution is passed through a QMA column, and the resulting solution is subjected to column chromatography18F]F-Adsorbed on QMA column.
2. Aspirate 1mL K222(0.5mg)/K2CO3(1mg)MeCN/H20-5/2 solution through 0.5M NaHCO3An activated QMA column of [2 ], [18F-]Eluting from the column to the reaction tube. The eluate of appropriate activity was taken to dryness at 100 ℃ under a nitrogen purge.
3. Adding into a reaction tubeAdding 1mL of anhydrous acetonitrile, evaporating to dryness, and repeating the evaporation process 3 times to obtain dried [ alpha ], [ alpha ] form18F-]F-/K222/K2CO3The complex was capped and sealed and then cooled.
4. 0.05mg of the labeled precursor was dissolved in 1mL of acetonitrile, vortexed and added to the above dried and cooled complex. The reaction was carried out at room temperature for 1min, and the reaction mixture was cooled after the reaction was completed.
5. The cooled reaction solution was diluted with 9mL of water, the diluted solution was purified by Oasis solid phase extraction column for intermediate purification and by washing with 10mL of water, and finally by rinsing the column with 1mL of ethanol and by HPLC purification (Phenomenex Gemini-Nx C18110A (250 × 4.6mm), 0.1% formic acid/acetonitrile 10% -90%: 35/75, with a peak time of 9.4min and a purity of greater than 95%.
EXAMPLE 19(((S) -1-carboxy-5- (4- ((((fluoro-18F) sulfonyl) oxy) benzamido) pentyl) carbamoyl) -L-glutamic acid
The structural formula is as follows:
the reaction formula is as follows:
1. suction contains18F]H of (A) to (B)2 18The O (20mCi) solution is passed through a QMA column, and the resulting solution is subjected to column chromatography18F]F-Adsorbed on QMA column.
2. Aspirate 1mL K222(0.5mg)/K2CO3(1mg)MeCN/H2o-5/2 solution through 0.5M NaHCO3An activated QMA column of [2 ], [18F-]Eluting from the column to the reaction tube. The eluate of appropriate activity was taken to dryness at 100 ℃ under a nitrogen purge.
3. Adding 1mL of anhydrous acetonitrile into the reaction tube, evaporating to dryness, and repeating the evaporation process for 3 times to obtainDried [ alpha ], [ beta ]18F-]F-/K222/K2CO3The complex was capped and sealed and then cooled.
4. 0.05mg of the labeled precursor was dissolved in 1mL of acetonitrile, vortexed and added to the above dried and cooled complex. The reaction was carried out at room temperature for 1min, and the reaction mixture was cooled after the reaction was completed.
5. The cooled reaction solution was diluted with 9mL of water, the diluted solution was purified by Oasis solid phase extraction column for intermediate purification and by washing with 10mL of water, and finally by rinsing the column with 1mL of ethanol and by HPLC purification (Phenomenex Gemini-Nx C18110A (250 × 4.6mm), 0.1% formic acid/acetonitrile 20% -80% ═ 35/75, the peak time was 5.3min, and the purity was greater than 95%.
EXAMPLE 20 binding experiments for (S) -4- ((2- (2-cyano-4, 4-difluoropyrrolidin-1-yl) -2-oxoethyl) carbamoyl) quinolin-6-yl thiofluoroacid ester
Three sets of 12 test tubes (parallel experiments), numbered 1-12 in sequence, to which an equivalent amount of known radioligand [2 ]125I]IDAM(0.1-0.2nM，Kd0.2 nM); in test tubes No. 2 to 11, 50. mu.L of a solution of the active ingredient was added to the test tubes from low to high (10)-10to 10-5M) Tris-HCl solution of the sample to be tested (diaryl sulfide derivative); adding 50 mu L of Tris-HCl solution without a sample to be detected into a test tube No. 1 as a blank control, and adding 0.1mg/mL citalopram Tris-HCl solution into a test tube No. 12 to determine the nonspecific binding with SERT; respectively adding 100 mu l of LLC-serotonin transporter (SERT) homogenate solution diluted by Tris-HCl buffer solution into test tubes from No. 1 to No. 12 for competitive reaction for 1 hour at room temperature; collecting cell membrane protein on a glass filter membrane (the filter membrane is soaked by 1% polyethyleneimine, the smooth surface is upward, and the hair surface is downward), washing the test tube and the filter membrane by using cold Tris-HCl buffer solution to remove non-specific binding, taking down the filter membrane, and placing the filter membrane into a gamma counter for measurement; the measured data is processed with Excel to obtain the concentration and [2 ] of the compound to be measured125I]Obtaining the compound IC to be tested by IDAM radioactive counting curve diagram50The value is calculated by a formula to obtain (S) -4- ((2- (2-cyano-4, 4-difluoropyrrolidine-1-)Base) -2-oxoethyl) carbamoyl) quinolin-6-yl thiofluoroacid ester had a Ki value of 0.4nM with SERT. The result shows that the compound has high affinity SERT and is expected to be applied to SERT related diseases such as: diagnosis of depression.
Example 21(2R, 3R, 11bR) -2-hydroxy-3-isobutyl-10-methoxy-11 b-methyl-1, 3,4,6,7,11 b-hexahydro-2H-pyridinyl [2,1-a ]]Isoquinoline-fluoro 9-18F
The rat brain striated tissue homogenate was stored at-80 ℃ in 50mM Hepes and 0.32M Sucrose solution (pH 7.5); three groups of 12 tubes (parallel experiments) are taken, numbered 1-12 in sequence, and an equal amount of known radioligand (2R, 3R, 11bR) -2-hydroxy-3-isobutyl-10-methoxy-11 b-methyl-1, 3,4,6,7,11 b-hexahydro-2H-pyridyl [2,1-a ] is added into the tubes]Isoquinoline-fluoro 9-18F (0.15-0.2 nM); in test tubes No. 2 to 11, 50. mu.L of a solution of the active ingredient was added to the test tubes from low to high (10)-12to 10-7M) (TBZ derivative) 50mM Hepes/0.32M sucrose (pH 7.5) solution; to test tube No. 1, 50 μ L of 50mM Hepes/0.32M sucrose (pH 7.5) solution containing no sample to be tested was added as a blank, and to test tube No. 12, 50 μ L of 50mM Hepes/0.32M sucrose (pH 7.5) solution containing an inhibitor was added for determination of nonspecific binding to VMAT 2; in test tubes No. 1 to No. 12, 100. mu.l of a homogenate of rat brain striatum tissue from 50mM Hepes/0.32M sucrose (pH 7.5) was added to each test tube and subjected to a competitive reaction at room temperature for 90 min; collecting cell membrane proteins on a glass filter membrane by using a cell collector, washing the test tube and the filter membrane by using a cold PBS buffer solution (pH is 4) to remove non-specific binding, taking down the filter membrane, and putting the filter membrane into a gamma counter for measurement; processing the measured data by Excel to obtain a compound IC to be measured50The value of (2R, 3R, 11bR) -2-hydroxy-3-isobutyl-10-methoxy-11 b-methyl-1, 3,4,6,7,11 b-hexahydro-2H-pyridinyl [2,1-a ] is calculated by the Cheng-Prusoff equation]Isoquinoline-fluoro 9-18The Ki value of F to receptor was 1.52 nM. From the results, the compound has higher affinity to VMAT2, and is expected to be applied to VMAT2 related diseases such as: and (4) diagnosis of Parkinson's disease.
EXAMPLE 22 affinity of examples 7-12 for fibroblast activation protein
The enzymatic activities of DPPIV, DPP8, DPP9, FAP and PREP were measured at 25 ℃ and fluorescence was monitored at an excitation wavelength of 380nm and an emission wavelength of 460 nm. The substrate for FAP analysis was Z-Gly-Pro-AMC. The reaction mixture contained 25. mu.M enzyme buffer (FAP) and appropriate amount of inhibitor (between 10)-4And 10-11M) in total, the total volume was 210 μ L. The final enzyme concentrations for FAP were 0.1, 0.8, 0.4, 1.2 and 0.6nM, respectively. The IC50 value is defined as the concentration of inhibitor required to reduce the enzyme activity by 50% after a 10 minute pre-incubation with the enzyme at 25 ℃ before addition of substrate. Examples 7-10 had IC50 values of 3.2,2.5,3.9,5.9,4.3, 4.6 nM. As can be seen from the results, examples 7 to 12 high affinity fibroblast activation protein allow diagnosis of fibroblast activation protein-associated tumors.
EXAMPLE 23 affinity of examples 13-15 to prostate specific membrane antigen protein
LNCaP cells (approximately 2X 10 per well)5) Inoculated into 6-well cell cultures and the dishes were loaded 48 hours prior to the experiment. After washing, cells were incubated with 37kBq example 15 at 37 ℃ and 4 ℃ for 5, 15, 30, 60, 90 and 120 minutes, respectively. Specific cellular uptake was determined by competitive blocking with 20 μ M2-phosphonomethylglutaric acid. Cellular uptake was terminated by washing twice with 4mL ice-cold PBS. Cells were then incubated twice with 0.5mL glycine-HCl in PBS (50mM, pH 2.8) for 5 minutes to remove surface binding. Cells were washed with 1mL ice-cold PBS and lysed with 0.5mL 1N NaOH. The radioactivity of the probe was measured in a gamma counter. Cellular uptake was calculated as binding to 106Percentage of radioactivity initially added to individual cells [% ID/10 [% ]6Cells]. Examples 13-15 had IC50 values of 8.5, 4.3,3.8 nM. From the results, it can be seen that example 15, a high affinity prostate specific membrane antigen protein, can diagnose PSMA-related diseases such as prostate cancer.
Example 2418F-labeling tumor cell uptake assays of examples 1-4
Cells were seeded in well plates 24h before the experiment: the culture was decanted, washed twice with PBS (2mL x 2), digested with 0.25% pancreatin (1mL), and blown down evenly. The pipette gun removed 50. mu.L of the single cell suspension, added 950. mu.L of the culture medium, and diluted 20-fold. Taking 10 muL was counted using a hemacytometer and the four large lattices total N. And (3) calculating: n/4 x 20 x 10 ═ N4one/mL. Diluting to 4 x 10 according to the calculated result5one/mL. The well plates were washed with PBS and rinsed with culture medium. After washing, the breast cancer MCF-7 cell line is washed at 2X 10 per well5The cells/well/4 groups (4 wells per time point in each group) were placed in 24-well plates, and 0.5mL of high-glucose DMEM low/1640 medium containing 10% Fetal Bovine Serum (FBS) was incubated in the incubator overnight at 37 ℃ with 5% CO2And performing wet culture for 24 h. At 30min and 120min, the uptake (% uptake/100ug protein) of the following compounds is shown in Table 1. As can be seen from Table 1, tumor pairs18F marked examples 1-4 all have higher uptake, and the compounds are expected to be applied to the diagnosis of tumors.
TABLE 1
Name of Compound
Uptake value in 30min
60 minute uptake value
18Labeling example 1 with F
15.3％
34.2％
18Labeling example 2 with F
16.4％
44.1％
18F Label example 3
13.2％
28.1％
18F Label example 4
14.9％
30.4％
Example 2518F-labelling of tumor biodistribution of examples 1-4,7-11
In order to study the biodistribution and pharmacokinetics of the compounds, examples 1-4(MCF-7 bearing mice), 7-10(HT-1080-FAP), 13(22rv1 bearing mice) in the patent were evaluated by in vivo distribution experiments in nude mice with different tumors. The radioactive arginine derivative was diluted to-300. mu. Ci/mL (. about.11 MBq/mL) with physiological saline. 0.1mL of the diluted solution was taken out, 9.9mL of physiological saline was added, and after mixing, 0.1mL of the diluted solution was taken out again as a standard solution (1%) whose radioactive count was measured simultaneously with the measurement of the tissue radioactive count.
In the case of MCF-7 tumor nude mice, the mice weighed 16 + -2 g, and 5 mice were randomized. 0.1mL (-30. mu. Ci,. -1 MBq) of arginine solution was injected from the tail vein, sacrificed at decapitation at 5, 30, 60, 120min after injection, blood and tumor were collected, wiped and weighed, radioactive counts were measured, and the uptake (% ID/g) of each organ and tissue and the ratio of tumor/muscle and tumor/blood were calculated. The results are shown in Table 2 below.
TABLE 2
As can be seen from the table, all compounds have a high tumor to muscle ratio and a long retention time of the imaging agent in the tumor. And the blood in the body is cleared quickly, the imaging is clear, and the method is suitable for tumor imaging.
Example 2618PET imaging of F-labeled example 1
Under isoflurane anesthesia (2-3%, 1L/min oxygen) and heating plate warming condition, the composition contains18Physiological saline of the F arginine derivative was injected into MCF-7 tumor nude mice, and then image acquisition was started. All the operation completion time is not less than 2h (dynamic, 5 min/frame), and the image is analyzed and reconstructed by AMIDE' S medical imaging software. As can be seen from the view of figure 1,18the F-tag example 1 can be used for tumor imaging, where the tumor is within the white circle.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.