Benzoheterocycle derivative containing diphosphonic acid / dicarboxylic acid group and application thereof

Abstract

The invention relates to the field of medicinal chemistry and radiopharmaceuticals, in particular to a benzoheterocycle derivative containing a diphosphonic acid / dicarboxylic acid group and application thereof, and the benzoheterocycle derivative has a structure shown in a general formula fI). After the compound and the derivative thereof are labeled by radionuclide, transthyroxine amyloidosis can be developed. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the fields of medicinal chemistry and radiopharmaceuticals, and in particular to benzoheterocyclic derivatives containing bisphosphonic acid / biscarboxylic acid groups and applications thereof in the diagnosis of transthyretin amyloidosis. Background Art

[0002] Transthyretin amyloidosis (ATTR) results from the dissociation and misfolding of the transthyretin protein. ATTR progresses slowly and has a long survival period, with a median survival of approximately three years after diagnosis. Immunoglobulin light chain amyloidosis (AL) arises from the expansion of clonal plasma cells that overproduce immunoglobulin light chains. It is a rapidly progressive disease with a high mortality rate, with a median survival of only six months. Although the clinical manifestations of the two conditions overlap significantly, their distinct pathophysiological substrates lead to distinct clinical courses, treatment approaches, and prognoses. In recent years, several drugs for the treatment of ATTR (patisiran, inotersen, tafamidis meglumine, tafamidis, vutrisiran, and eplontersen) have been approved by the Food and Drug Administration (FDA). These drugs have proven to be effective in improving disease progression, but misdiagnosis or delayed diagnosis can miss the opportunity for timely treatment. Therefore, developing diagnostic methods to differentiate ATTR from AL is of great clinical, therapeutic, and prognostic significance.

[0003] Radioactive bone tracers have been shown to be crucial in the diagnosis of ATTR. Studies have found that the number of microcalcifications in ATTR is significantly greater than in AL, and the bisphosphonate group in the bone imaging agent can bind to calcium, so radioactive bone tracers can specifically diagnose ATTR. In 2016, the results of a multicenter clinical study conducted by Gillmore et al. showed that myocardial uptake combined with blood, urine immunofixation electrophoresis and serum free light chain detection can accurately diagnose ATTR-type cardiac amyloidosis (CA) with high sensitivity and specificity. Confirmed 99m The value and status of Tc-PYP / DPD / HMDP cardiac imaging in the non-invasive diagnosis of ATTR-CA. However, the visual grading system and quantitative parameters of bone imaging still need to be further improved and standardized. In addition, the resolution of SPECT imaging is low and cannot be absolutely quantitative, which limits the application of bone imaging agents in the diagnosis of ATTR. 124 I] Evuzamitideu is the only PET diagnostic imaging agent to receive breakthrough therapy designation for cardiac amyloidosis. However, compared with the reported β-amyloid tracers [ 18 F] Florbetapir similar, [ 124I] Evuzamitideu does not have the ability to distinguish between ATTR and AL. Therefore, the development of PET probes that can distinguish AL and ATTR has important scientific significance and clinical value. Summary of the Invention

[0004] The purpose of the present invention is to provide a preparation method and application of a molecular probe that has high affinity and selectivity for ATTR.

[0005] In a first aspect, the present application provides a compound of formula (I),

[0006]

[0007] in

[0008] X1 is selected from S or O; X2 is selected from N or CH;

[0009] When R1 is fluorinated oligoethylene glycol, R2=R3, and is selected from the group consisting of: -C 1-4 Alkyl-carboxyl, -C 1-4 Alkyl-bisphosphonic acid group;

[0010] When R1 is When R2 and R3 are each independently selected from -CH3 or -H, wherein R4 is selected from the group consisting of: -C 1-10 Alkyl, -oligoethylene glycol; R5 is selected from the group consisting of: -C 1-10 Alkyl-chelate group, -oligoethylene glycol-chelate group; R6 is selected from the group consisting of: -C 1-10 Alkyl-bisphosphonic acid group, -oligoethylene glycol-bisphosphonic acid group; wherein the oligoethylene glycol is a oligoethylene glycol composed of 1-5 ethylene glycols; fluorinated oligoethylene glycol means that the terminal hydroxyl group of the oligoethylene glycol is substituted by fluorine;

[0011] wherein the bisphosphonic acid group is

[0012] The chelating group is a chelating radionuclide 68 Ga, 64 Cu, Al 18 F. 99 Chelating group for mTc or the corresponding stable isotope.

[0013] In some embodiments, when R1 is fluoro-oligoethylene glycol, R2=R3, and is selected from the group consisting of: -C 1-4 Alkyl-carboxyl, -C 1-4Alkyl-bisphosphonic acid group; preferably, R1 is -(CH2CH2O)2CH2CH2F, and R2=R3 is selected from the group consisting of: -CH2CH2COOH, -CH2COOH,

[0014] In some embodiments, when R1 is When R2 and R3 are each independently selected from -CH3 or -H, wherein R4 is selected from the group consisting of: -C 1-10 Alkyl, -oligoethylene glycol; R5 is selected from the group consisting of: -C 1-10 Alkyl-chelate group, -oligoethylene glycol-chelate group; R6 is selected from the group consisting of: -C 1-10 Alkyl-bisphosphonic acid group, -oligoethylene glycol-bisphosphonic acid group; preferably, R4 is -(CH2CH2O)2CH2CONH-; R5 is selected from And R6 is Where M is 68 Ga 3+ 、 67 Ga 3+ or its corresponding stable isotope.

[0015] In some embodiments, the compound of formula (I) is selected from the group consisting of:

[0016]

[0017]

[0018] The second embodiment of the present application provides a radioactive labeled compound of the compound as described in any embodiment of the first aspect, wherein the radioactive labeled compound is prepared by reacting the compound as described in any embodiment of the first aspect. 18 F, 68 Ga or 67 Ga labeled.

[0019] The third aspect of the present application provides a pharmaceutical composition comprising the compound as described in any embodiment of the first aspect or the radiolabeled compound as described in any embodiment of the second aspect; and a pharmaceutically acceptable carrier.

[0020] The fourth aspect of the present application provides a diagnostic or detection reagent, comprising a compound as described in any embodiment of the first aspect or a radiolabeled compound as described in any embodiment of the second aspect; and a pharmaceutically acceptable carrier.

[0021] In some embodiments, the diagnostic or detection reagent is used to diagnose or detect transthyretin amyloidosis.

[0022] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0023] The benzoheterocyclic derivatives containing bisphosphonic acid / biscarboxylic acid groups provided by the present invention have good affinity and selectivity for ATTR aggregates and microcalcifications thereof, and can be used for the diagnosis of ATTR patients after being labeled with radioactive isotopes. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0025] Figure 1 This is the synthetic route of compounds 15 and 16.

[0026] Figure 2 This is the synthetic route of compound 24.

[0027] Figure 3 This is the synthetic route for compounds 49, 50 and 51.

[0028] Figure 4 Ca 2+ The effects of compound 50 on Ca at different molar ratios with compound 50 2+ The calcium chelation rate was 1.0477 W / m and 1.571 W / m, respectively. EDTA was used as a positive control.

[0029] Figure 5 for[ 18 F]16, [ 68 Ga]Ga-49 and [ 68 Autoradiographic results of Ga]Ga-50 in brain sections of Aβ patients, myocardial sections of ATTR patients, AL patients and healthy volunteers.

[0030] Figure 6 for[ 68 Dynamic PET / CT imaging results of Ga]Ga-50 in SD rats, Figure 6 (A) PET / CT fusion images at different time points (from left to right: transverse, sagittal, and coronal planes) and the maximum signal intensity projection (MIP) of the whole-body transverse section at 60-70 minutes. Figure 6 (B) is the time-activity curve (TAC) in different organs. Figure 6 (C) Heart-to-lung and heart-to-liver uptake ratios at different times.

[0031] Figure 7 for[68 Dynamic PET / CT imaging results of Ga]Ga-49 in SD rats, Figure 7 (A) PET / CT fusion images at different time points (from left to right: transverse, sagittal, and coronal planes) and the maximum signal intensity projection (MIP) of the whole-body transverse section at 60-70 minutes. Figure 7 (B) is the time-activity curve (TAC) in different organs. Figure 7 (C) Heart-to-lung and heart-to-liver uptake ratios at different times. DETAILED DESCRIPTION

[0032] To facilitate understanding of the present invention, the following further describes the implementation process of the present invention in conjunction with specific implementation examples. These descriptions are only intended to further illustrate the features and advantages of the present invention and are not intended to limit the claims of the invention. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are intended to fall within the scope of protection of the present invention.

[0033] Example 1: Synthesis of Compound 1

[0034]

[0035] Under a nitrogen atmosphere at 0°C, 4-benzyloxyaniline (100 mg, 0.5 mmol) and ammonium thiocyanate (114 mg, 1.5 mmol) were dissolved in glacial acetic acid containing 20% formic acid. Br2 (20 μL, 1.5 mmol) was added dropwise in a dark atmosphere. The dark atmosphere was removed and the reaction was allowed to react at room temperature overnight. After completion of the reaction, 1 M NaOH was added at 0°C to adjust the pH to 11. The mixture was dissolved in ethyl acetate and filtered through Celite. The filtrate was washed sequentially with saturated NaHCO3 solution, saturated brine, and water, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and separated by column chromatography (PE / EA = 1 / 1, v / v) to afford a white solid (85 mg, 66%). 1 H NMR (400MHz, CDCl3) δ7.48-7.31 (m, 6H), 7.20 (d, J=2.5Hz, 1H), 6.99 (dt, J=8.8, 2.5Hz, 1H), 5.08 (s, 2H).MS: m / z calcd.for[C 14 H 12 N2OS+H] + 257.1, found 257.2.

[0036] Example 2: Synthesis of Compound 2

[0037]

[0038] Compound 1 (1000 mg, 3.9 mmol) was dissolved in 50% KOH solution (20 mL), and ethylene glycol (5 mL) was added. The mixture was refluxed overnight. After completion of the reaction, 30 mL of toluene was added, and the mixture was adjusted to neutral with glacial acetic acid. The mixture was washed with water, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and separated by column chromatography (PE / EA = 2 / 1, v / v) to obtain a green solid (626 mg, 70%). 1 H NMR (600MHz, CDCl3) δ7.34 (d, J=4.5Hz, 4H), 7.30 (dd, J=5.4, 3.1Hz, 1H), 6.88 (dd, J =8.7, 2.9Hz, 1H), 6.81 (d, J = 2.9Hz, 1H), 6.68 (d, J = 8.7Hz, 1H), 4.81 (s, 2H).MS: m / z calcd.for[C 14 H 17 NOS+H] + 231.1, found 231.2.

[0039] Example 3: Synthesis of Compound 3

[0040]

[0041] Aniline (465 mg, 5.0 mmol), methyl acrylate (1291 mg, 15 mmol), hydroquinone (55 mg, 0.5 mmol), and acetic acid (3 mL) were added to a 100 mL round-bottom flask and reacted at 100°C for 20 h. After completion of the reaction, the mixture was concentrated under reduced pressure and separated by column chromatography (PE / EA = 8 / 1, v / v) to afford a red oil (600 mg, 45%). 1 H NMR (400MHz, CDCl3) δ7.24 (td, J=7.3, 1.5Hz, 2H), 6.76-6.69 (m, 3H), 3.67 (m, 10H), 2.62-2.57 (m, 4H).MS: m / zcalcd.for[C 14 H 19 NO4+H] + 266.1, found 266.2.

[0042] Example 4: Synthesis of Compound 4

[0043]

[0044] Aniline (186 mg, 2.0 mmol) was dissolved in 10 mL of acetonitrile, and KCO (636 mg, 4.6 mmol) and KI (622 mg, 4.5 mmol) were added. The mixture was allowed to react at room temperature for 10 minutes, followed by the addition of ethyl bromoacetate (752 mg, 4.5 mmol). The reaction was allowed to proceed at 90°C overnight. After completion of the reaction, the mixture was concentrated under reduced pressure, dissolved in dichloromethane, washed with water, dried over anhydrous NaSO, filtered, concentrated under reduced pressure, and separated by column chromatography (PE / EA = 10 / 1, v / v) to afford a brown oil (209 mg, 39%). 1 H NMR (600MHz, CDCl3) δ7.22 (dd, J=8.7, 7.4Hz, 2H), 6.79 (t, J=7.3Hz, 1H), 6.65- 6.61 (m, 2H), 4.22 (q, J=7.1Hz, 4H), 4.14 (s, 4H), 1.28 (t, J=7.1Hz, 6H). MS: m / z calcd.for[C 14 H 19 NO4+H] + 266.1, found 266.2.

[0045] Example 5: Synthesis of Compound 5

[0046]

[0047] Compound 3 (600 mg, 2.3 mmol) was dissolved in 5 mL of DMF, and POCl (388 mg, 2.5 mmol) was slowly added dropwise. The reaction was allowed to proceed at room temperature for 30 min, and then at 90°C for 2 h. After completion of the reaction, the mixture was cooled to room temperature and ice water was added. Sodium acetate was added to adjust the pH to neutral. The mixture was extracted with EA and washed with water. The organic layer was dried over anhydrous NaSO, filtered, concentrated under reduced pressure, and separated by column chromatography (PE / EA = 2 / 1, v / v) to obtain a red oil (329 mg, 45%). 1 H NMR (600MHz, CDCl3) δ9.75 (s, 1H), 7.74 (d, J=8.9Hz, 2H), 6.72 (d, J=9.0Hz, 2H), 3.78-3.75 (m, 4H), 3.69 (s, 6H), 2.65-2.62 (m, 4H).MS: m / zcalcd.for[C 15 H 19 NO5+H] + 294.1, found 294.2.

[0048] Example 6: Synthesis of Compound 6

[0049]

[0050] According to the synthesis method of compound 5, a yellow solid (122 mg, 55%) was obtained by column chromatography (PE / EA=2 / 1, v / v). 1 H NMR (400MHz, CDCl3) δ9.77 (s, 1H), 7.73 (d, J=8.9Hz, 2H), 6.65 (d, J=8.9Hz, 2H), 4.26-4.17 (m, 8H), 1.27 (t, J=7.1Hz, 6H). MS: m / z calcd.for[C 15 H 19 NO5+H] + 294.1, found294.2.

[0051] Example 7: Synthesis of Compound 7

[0052]

[0053] Compound 5 (330 mg, 1.1 mmol) and compound 2 (260 mg, 1.1 mmol) were weighed and dissolved in DMSO (5 mL), reacted at 170°C for 20 min, added with water, extracted with EA, washed with water, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography (PE / EA = 3 / 1, v / v) to obtain a black solid (404 mg, 80%). 1 H NMR (600MHz, CDCl3) δ7.92-7.86 (m, 3H), 7.45 (d, J=6.7Hz, 2H), 7.41-7.36 (m, 3H), 7.33 (d, J=6.6Hz, 1H), 7.11 (d, J=8.8Hz, 1H), 6.72 (d, J=7.8Hz, 2H), 5.09 (s, 2H), 3.72 (s, 4H), 3.69 (s, 6H), 2.62 (s, 4H). MS: m / z calcd.for[C 28 H 28 N2O5S+H] + 505.2, found 505.3.

[0054] Example 8: Synthesis of Compound 8

[0055]

[0056] According to the synthesis method of compound 7, a yellow-green solid (122 mg, 55%) was obtained by column chromatography (PE / EA=2 / 1, v / v). 1H NMR (400MHz, CDCl3) δ7.90 (t, J=9.3Hz, 3H), 7.44 (d, J=7.1Hz, 2H), 7.41-7.35 (m, 3H), 7.33 (d, J=7.1Hz, 1H), 7. 12 (dd, J=8.9, 2.3Hz, 1H), 6.68 (d, J=8.8Hz, 2H), 5.07 (s, 2H), 4.26-4.18 (m, 8H), 1.28 (t, J=7.1Hz, 6H). MS: m / z calcd.for[C 28 H 28 N2O5S+H] + 505.2, found 505.3.

[0057] Example 9: Synthesis of Compound 9

[0058]

[0059] -78 ° C. Under N2 atmosphere, compound 7 (403 mg, 0.8 mmol) was dissolved in anhydrous DCM, and BBr3 (1 M) (2.4 mL, 2.4 mmol) was slowly added and reacted for 1 h. After the reaction was completed, water was added to quench the reaction, and 1 M NaOH solution was adjusted to neutral. The mixture was extracted with DCM, washed with water, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography (PE / EA = 1 / 1, v / v) to obtain a brown solid (287 mg, 87%). 1 H NMR (400MHz, CDCl3) δ7.86 (d, J=8.9Hz, 2H), 7.77 (d, J=8.8Hz, 1H), 7.28 (d, J=2.4Hz, 1H), 6.96 (dd, J=8 .8, 2.4Hz, 1H), 6.69 (d, J=9.0Hz, 2H), 3.72 (t, J=7.1Hz, 4H), 3.68 (s, 6H), 2.61 (d, J=7.0Hz, 4H). MS: m / z calcd.for[C 21 H 22 N2O5S+H] + 415.1, found 415.3.

[0060] Example 10: Synthesis of Compound 10

[0061]

[0062] According to the synthesis method of compound 9, a yellow solid (625 mg, 85%) was obtained by column chromatography (PE / EA=1 / 1, v / v). 1H NMR (600MHz, CDCl3) δ7.83 (d, J=8.7Hz, 1H), 7.79 (d, J=8.8Hz, 2H), 7.06 (d, J=2.3Hz, 1H), 6.93 (dd, J=8 .8, 2.5Hz, 1H), 6.56 (d, J=9.0Hz, 2H), 4.26 (q, J=7.1Hz, 4H), 4.19 (s, 4H), 1.31 (t, J=7.1Hz, 6H).MS: m / z calcd.for[C 21 H 22 N2O5S+H] + 415.1, found415.3.

[0063] Example 11: Synthesis of Compound 11

[0064]

[0065] Compound 9 (70 mg, 0.2 mmol), triethylene glycol di(p-toluenesulfonate) (115 mg, 0.3 mmol), K2CO3 (138 mg, 1.0 mmol) and 18-crown ether-6 (catalytic amount) were added to acetone (10 mL) and refluxed for 4 h. After the reaction was completed, the solvent was removed by rotary evaporation, dissolved in DCM, filtered, concentrated under reduced pressure, and purified by column chromatography (PE / EA=1 / 1, v / v) to obtain a yellow solid (60 mg, 50%). 1 H NMR (600MHz, CDCl3) δ7.90 (d, J=8.8Hz, 2H), 7.85 (d, J=8.9Hz, 1H), 7.78 (d, J= 8.3Hz, 2H), 7.36-7.27 (m, 3H), 7.04 (dd, J=8.8, 2.6Hz, 1H), 6.72 (d, J=9.0Hz, 2H), 4.19-4.12(m, 4H), 3.89-3.81(m, 2H), 3.73(t, J=7.2Hz, 4H), 3.69(m, 8H) , 3.67-3.66(m, 2H), 3.64-3.60(m, 2H), 2.67-2.60(m, 4H), 2.40(s, 3H).MS: m / z calcd.for[C 34 H 40 N2O 10 S2+H] + 701.2, found 701.3.

[0066] Example 12: Synthesis of Compound 12

[0067]

[0068] According to the synthesis method of compound 11, a yellow solid (53 mg, 63%) was obtained by column chromatography (PE / EA=1 / 1, v / v). 1 H NMR (400MHz, CDCl3) δ7.91 (d, J=8.9Hz, 2H), 7.87 (d, J=8.9Hz, 1H), 7.78 (d, J=8.3Hz, 2 H), 7.33-7.31 (m, 2H), 7.29 (s, 1H), 7.05 (dd, J=8.9, 2.5Hz, 1H), 6.69-6.65 (m, 2H), 4.2 3 (q, J=7.1Hz, 4H), 4.19 (s, 4H), 4.18-4.14 (m, 4H), 3.85 (dd, J=5.5, 4.0Hz, 2H), 3.72-3 .66 (m, 4H), 3.64-3.60 (m, 2H), 2.40 (s, 3H), 1.29 (t, J=7.1Hz, 6H).MS: m / zcalcd.for[C 34 H 40 N2O 10 S2+H] + 701.2, found 701.3.

[0069] Example 13: Synthesis of Compound 13

[0070]

[0071] According to the synthesis method of compound 11, a yellow solid (145 mg, 88%) was obtained by column chromatography (PE / EA=1 / 1, v / v). 1 H NMR (600MHz, CDCl3) δ7.87 (d, J=8.9Hz, 2H), 7.83 (d, J=8.9Hz, 1H), 7.30 (d, J=2.4Hz, 1H), 7.03 (dd, J=8.9, 2.5Hz, 1H), 6.70 (d, J=9.0Hz, 2H), 4.58-4. 55(m, 1H), 4.50-4.47(m, 1H), 4.17-4.14(m, 2H), 3.87-3.84(m, 2H), 3.75- 3.73 (m, 1H), 3.73-3.68 (m, 9H), 3.66 (s, 6H), 2.60 (t, J=7.2Hz, 4H).MS: m / z calcd.for[C 27 H 33 FN2O7S+H] + 549.2, found549.4.

[0072] Example 14: Synthesis of Compound 14

[0073]

[0074] According to the synthesis method of compound 11, a yellow solid (625 mg, 95%) was obtained by column chromatography (PE / EA=1 / 1, v / v). 1 H NMR (600MHz, CDCl3) δ7.92 (d, J=8.6Hz, 2H), 7.89 (d, J=8.8Hz, 1H), 7.33 (d, J= 2.5Hz, 1H), 7.07 (dd, J=8.9, 2.5Hz, 1H), 6.68 (d, J=8.9Hz, 2H), 4.61-4.60 (m, 1H), 4.54-4.52 (m, 1H), 4.24 (q, J=7.1Hz, 4H), 4.20 (d, J=6.6Hz, 6H), 3.91-3. 89 (m, 2H), 3.79-3.75 (m, 3H), 3.75-3.72 (m, 3H), 1.29 (t, J=7.1Hz, 6H).MS: m / z calcd.for[C 27 H 33 FN2O7S+H] + 549.2, found 549.4.

[0075] Example 15: Synthesis of Compound 15

[0076]

[0077] Compound 13 (110 mg, 0.2 mmol) was dissolved in ethanol (6 mL), and KOH (34 mg, 0.6 mmol) was added. The mixture was reacted at 60°C for 1 h. After the reaction was completed, the pH was adjusted to 3 with 1 M hydrochloric acid solution, and an off-white solid (70 mg, 75%) was obtained by filtration. 1 H NMR (600MHz, DMSO-d6) δ7.82 (dd, J=8.7, 6.2Hz, 3H), 7.63 (d, J=2.5Hz, 1H), 7.07 (dd, J=8.9, 2.5Hz, 1H), 6.81 (d, J=8.8Hz, 2H), 4.57-4.54 (m, 1H ), 4.49-4.45 (m, 1H), 4.17 (t, J=4.6Hz, 2H), 3.78 (dd, J=5.6, 3.5Hz, 2H), 3.70-3.61 (m, 8H), 3.59 (dd, J=6.0, 3.5Hz, 2H), 2.53 (t, J=7.2Hz, 4H). 13C NMR (101MHz, DMSO-d6) δ173.61, 165.79, 156.54, 149.41, 148.90, 135.75, 128.96, 122.99, 121.19, 116.1 1, 112.28, 106.23, 83.58 (d, J=165.6Hz), 70.46, 70.38, 70.24 (d, J=19.1Hz), 69.47, 68.29, 46.70, 32.39. 19 F NMR(565MHz, DMSO-d6)δ-221.18.HRMS: m / z calcd for[C 25 H 29 FN2O7S-H] - 519.1607, found 519.1597.

[0078] Example 16: Synthesis of Compound 16

[0079]

[0080] Following the synthesis method of compound 15, a pale yellow solid (32 mg, 65%) was obtained by filtration. H NMR (400 MHz, DMSO-d6) δ 7.83 (dd, J = 8.9, 2.0 Hz, 3H), 7.65 (d, J = 2.4 Hz, 1H), 7.08 (dd, J = 8.9, 2.4 Hz, 1H), 6.67 (d, J = 8.9 Hz, 2H), 4.60-4.55 (m, 1H), 4.47-4.43 (m, 1H), 4.19 (d, J = 9.1 Hz, 6H), 3.81-3.76 (m, 2H), 3.72-3.68 (m, 1H), 3.64-3.58 (m, 5H). 13 C NMR (151MHz, DMSO-d6) δ172.25, 165.67, 156.63, 150.60, 148.85, 135.86, 128.65, 123.11, 122.25, 11 6.20, 112.40, 106.25, 83.58 (d, J=165.8Hz), 70.47, 70.38, 70.24 (d, J=18.9Hz), 69.47, 68.31, 53.35. 19 F NMR(376MHz, DMSO-d6)δ-221.60.HRMS: m / z calcd for[C 23 H 25 FN2O7S-H] - 491.1294, found 491.1285.

[0081] Example 17: Synthesis of Compound 17

[0082]

[0083] Under a nitrogen atmosphere, dibenzylamine (11.8 g, 20 mmol), triethoxymethane (10.6 g, 70 mmol) and diethyl phosphite (25.6 g, 186 mmol) were stirred at 160° C. for 24 h. During the reaction, the resulting ethanol was removed several times. After cooling, 300 mL of CHCl 3 was added to the system, and the mixture was washed with 5% NaOH and H 2 O in sequence, dried over anhydrous Na 2 SO 4 , and concentrated under reduced pressure to give a yellow oil (6.1 g, 63%) by column chromatography (PE / EA=1 / 1, v / v). 1 H NMR (600MHz, CDCl3) δ7.41 (s, 4H), 7.29 (t, J=7.4Hz, 4H), 7.23 (t, J=7.0Hz, 2H), 4.15 ( m, 8H), 4.11-4.02 (m, 4H), 3.55 (t, J=25.1Hz, 1H), 1.32 (dt, J=7.1, 3.6Hz, 12H).MS: m / z calcd.for[C 23 H 35 NO6P2+H] + 484.2, found484.1.

[0084] Example 18: Synthesis of Compound 18

[0085]

[0086] 10% Pd / C (106 mg, 1.0 mmol) was added to a mixture of compound 17 (483 mg, 1.0 mmol) and 10 mL of MeOH, stirred at room temperature overnight under a hydrogen atmosphere, filtered, and the filtrate was concentrated under reduced pressure to obtain a colorless oil (279 mg, 92%). 1 HNMR (600MHz, CDCl3) δ4.24-4.19 (m, 8H), 3.41 (t, J=20.6Hz, 1H), 1.34 (t, J=7.1Hz, 12H).MS: m / zcalcd.for[C9H 23 NO6P2+H] + 304.1, found 304.1.

[0087] Example 19: Synthesis of Compound 19

[0088]

[0089] According to the synthesis method of compound 15, an off-white solid (137 mg, 97%) was obtained by filtration. 1 H NMR (400MHz, DMSO-d6) δ7.83 (dd, J=8.9, 1.3Hz, 3H), 7.72 (d, J=2.6Hz, 1H), 7.51-7.47 (m, 2H), 7.43-7.38 (m, 2H), 7.35 (dd, J=6. 7, 2.0Hz, 1H), 7.14 (dd, J=8.9, 2.6Hz, 1H), 6.81 (d, J=9.1Hz, 2H), 5.18 (s, 2H), 3.65 (t, J=7.0Hz, 4H), 2.56-2.52 (m, 4H). MS: m / z calcd.for[C 26 H 24 N2O5S-H]-475.1, found475.0.

[0090] Example 20: Synthesis of Compound 20

[0091]

[0092] Compound 18 (400 mg, 0.8 mmol) was added to a mixed solution of compound 19 (612 mg, 2.0 mmol) and MeOH (10 mL), followed by the addition of DMT-MM (702 mg, 2.5 mmol). The mixture was allowed to react at room temperature for 6 h. The mixture was concentrated under reduced pressure, dissolved in ethyl acetate, washed sequentially with 1M HCl and water, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography. A brown solid (499 mg, 57%) was obtained by column chromatography (DCM / MeOH = 9 / 1, v / v). 1 H NMR (600MHz, CDCl3) δ7.88-7.84 (m, 3H), 7.43 (d, J=7.0Hz, 2H), 7.39-7.35 (m, 3H), 7.32 (d, J=5.0Hz, 1H), 7.10 (dd, J=8.9, 2.5Hz, 1H), 6.78 (d, J=9 .0Hz, 2H), 5.10 (s, 2H), 5.10-5.01 (m, 2H), 4.20-4.11 (m, 16H), 3.74 (t, J =7.1Hz, 4H), 2.61 (t, J = 7.1Hz, 4H), 1.29 (dd, J = 11.8, 7.0Hz, 24H). MS: m / z calcd.for[C 44 H 66 N4O 15 P4S+H] + 1047.3, found 1047.2.

[0093] Example 21: Synthesis of Compound 21

[0094]

[0095] According to the synthesis method of compound 18, a yellow oil (312 mg, 63%) was obtained. 1 H NMR (600MHz, CDCl3) δ8.68 (s, 1H), 7.76 (d, J=8.7Hz, 1H), 7.69 (d, J=8.5Hz, 2H), 7.12 (q, J=3.9Hz, 3H), 6.99 (dd, J=8.7, 2.5Hz, 1H), 6.70 (d, J =8.8Hz, 2H), 5.09 (td, J=21.9, 10.0Hz, 2H), 4.23-4.14 (m, 16H), 3.74 (t, J=7.0Hz, 4H), 2.60 (t, J=6.9Hz, 4H), 1.30 (dd, J=14.7, 7.3Hz, 24H).

[0096] Example 22: Synthesis of Compound 22

[0097]

[0098] According to the synthesis method of compound 11, a yellow solid (50 mg, 67%) was obtained by column chromatography (DCM / MeOH=9 / 1, v / v). 1 H NMR (400MHz, CDCl3) δ7.84 (dd, J=14.8, 8.9Hz, 3H), 7.76 (d, J=8.3Hz, 2H), 7.34-7.27 (m , 5H), 7.02 (dd, J=8.9, 2.5Hz, 1H), 6.79 (d, J=7.1Hz, 2H), 5.05 (td, J=22.0, 10.0Hz, 2H) , 4.19-4.09(m, 20H), 3.85-3.79(m, 2H), 3.76-3.71(m, 4H), 3.69-3.64(m, 4H), 3.61-3. 58(m, 2H), 2.60(t, J=7.0Hz, 4H), 2.38(s, 3H), 1.31-1.25(m, 24H).MS: m / zcalcd.for[C 50 H 78 N4O 20 P4S2+H] + 1243.4, found 1243.2.

[0099] Example 23: Synthesis of Compound 23

[0100]

[0101] According to the synthesis method of compound 11, a yellow solid (112 mg, 79%) was obtained by column chromatography (DCM / MeOH=9 / 1, v / v). 1 H NMR (400MHz, CDCl3) δ7.86-7.78 (m, 3H), 7.41 (d, J=10.1Hz, 2H), 7.29 (d, J=2.5 Hz, 1H), 7.01 (dd, J=8.9, 2.5Hz, 1H), 6.75 (d, J=9.1Hz, 2H), 5.03 (td, J=22.0, 1 0.0Hz, 2H), 4.60-4.55(m, 1H), 4.48-4.43(m, 1H), 4.16-4.07(m, 18H), 3.88-3. 82 (m, 2H), 3.71 (m, 10H), 2.59 (t, J=6.7Hz, 4H), 1.25 (q, J=7.2Hz, 24H).MS: m / z calcd.for[C 43 H 71 FN4O 17 P4S+H] + 1091.4, found 1091.2.

[0102] Example 24: Synthesis of Compound 24

[0103]

[0104] Compound 23 (142 mg, 0.1 mmol) was dissolved in 8 mL of dichloromethane, and trimethylsilyl bromide (153 mg, 1.0 mmol) was added dropwise at 0°C. The mixture was allowed to react at room temperature overnight. After concentration under reduced pressure, the mixture was dissolved in dichloromethane (8 mL), and trifluoroacetic acid (4 mL) was added dropwise. The reaction was allowed to react at room temperature for 3 h, and the mixture was concentrated under reduced pressure. The mixture was washed with ether to obtain a yellow solid (100 mg, 87%). 1 H NMR (400MHz, DMSO-d6) δ8.38 (d, J=10.2Hz, 2H), 7.82 (t, J=9.0Hz, 3H), 7.64 (s, 1H), 7.07 (d, J=9.8Hz, 1H), 6.82 (d, J=8.4Hz, 2H), 4. 68-4.52(m, 3H), 4.47-4.43(m, 1H), 4.20-4.14(m, 2H), 3.81-3.76(m, 2H), 3.72-3.67(m, 1H), 3.66-3.56(m, 9H), 2.58-2.52(m, 4H). 13C NMR (101MHz, DMSO-d6) δ170.58, 165.95, 156.51, 149.76, 148.80, 135.68, 128.92, 122.90, 120.80, 116.08, 1 12.12, 106.25, 83.58 (d, J=165.7Hz), 70.46, 70.37, 70.24 (d, J=19.4Hz), 69.47, 68.29, 65.46, 47.19, 33.69. 19 FNMR(376MHz, DMSO-d6)δ-221.18.HRMS: m / z calcd for[C 27 H 39 FN4P 17 P4S-H]-865.0893, found 865.0872.

[0105] Example 25: Synthesis of Compound 25

[0106]

[0107] According to the synthesis method of compound 20, a colorless oil (420 mg, 72%) was obtained by column chromatography (DCM / MeOH=9 / 1, v / v). 1 H NMR (400MHz, CDCl3) δ7.50 (d, J=9.8Hz, 1H), 7.14-7.23 (m, 5H), 6.70 (t, J=4.4Hz, 1H), 4.90-5.03 (m, 3H), 4.00- 4.11 (m, 8H), 3.95 (d, J=4.0Hz, 2H), 3.56 (s, 2H), 3.51 (d, J=4.0Hz, 4H), 3.26 (s, 2H), 1.10-1.22 (m, 12H).MS: m / z calcd.for[C 23 H 40 N2O 11 P2+H] + 583.4, found 583.1.

[0108] Example 26: Synthesis of Compound 26

[0109]

[0110] According to the synthesis method of compound 18, a yellow oil (435 mg, 97%) was obtained. 1H NMR (600MHz, CDCl3) δ8.37 (s, 2H), 7.81 (d, J=10.4Hz, 1H), 5.03 (td, J=21.5, 10.4Hz, 1H), 4.21-4.27 (m, 4H), 4.17-4.09 (m, 4H), 4.07 (s, 2H), 3.93 (s, 2H), 3.74-3.69 (m, 4H), 3.41 (s, 2H), 1.37 (t, J=6.9Hz, 6H), 1.31 (t, J=6.9Hz, 6H). MS: m / z calcd.for[C 15 H 34 N2O9P2+H] + 449.2, found449.3.

[0111] Example 27: Synthesis of Compound 27

[0112]

[0113] Tert-butyl bromoacetate (3900 mg, 20 mmol), 2-(2-chloroethoxy)ethanol (1246 mg, 10 mmol), TBAB (3223 mg, 10 mmol) and t-BuOK (1122 mg, 10 mmol) were added to 30 mL of THF and stirred at room temperature overnight. The mixture was concentrated under reduced pressure, dissolved in EA (50 mL), washed with H2O, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography (PE / EA=6 / 1, v / v) to obtain a brown oil (500 mg, 21%). 1 H NMR (600MHz, CDCl3) δ4.02 (s, 2H), 3.76 (t, J=5.9Hz, 2H), 3.71 (s, 4H), 3.63 (t, J=5.9Hz, 2H), 1.46 (s, 9H).MS: m / z calcd.for[C 10 H 19 ClO4+Na] + 261.1, found 261.1.

[0114] Example 28: Synthesis of Compound 28

[0115]

[0116] Compound 27 (419 mg, 1.8 mmol), 2-(4-methylaminophenyl)-6-hydroxybenzothiazole (300 mg, 1.2 mmol), KCO (486 mg, 3.5 mmol), and KI (332 mg, 2.0 mmol) were added to 10 mL of DMF and stirred at 95° C. overnight. After cooling, 50 mL of H O was added to the system, and the mixture was extracted with DCM, dried over anhydrous NaSO, filtered, concentrated under reduced pressure, and purified by column chromatography (PE / EA=2 / 1, v / v) to obtain a yellow oil (305 mg, 57%). 1 H NMR (400MHz, CDCl3) δ7.79 (dd, J=8.8, 6.1Hz, 3H), 7.24 (d, J=2.5Hz, 1H), 7.00 (dd, J=8.9, 2.5Hz, 1H), 6.54 (d, J=8 .7Hz, 2H), 4.12-4.07(m, 2H), 4.01(s, 2H), 3.84-3.80(m, 2H), 3.73-3.69(m, 4H), 2.78(s, 3H), 1.43(s, 9H).MS: m / z calcd.for[C 24 H 30 N2O5S+H] + 459.2, found 459.3.

[0117] Example 29: Synthesis of Compound 29

[0118]

[0119] According to the synthesis method of compound 28, a yellow solid (259 mg, 55%) was obtained by column chromatography (PE / EA=2 / 1, v / v). 1 H NMR (400MHz, CDCl3) δ7.90 (m, 3H), 7.34 (s, 1H), 7.06 (dd, J=8.9, 2.5Hz, 1H), 6.75 (d, J=8.9Hz, 2H), 4. 23-4.18(m, 2H), 4.05(s, 2H), 3.94-3.88(m, 2H), 3.80-3.74(m, 4H), 3.05(s, 6H), 1.47(s, 9H).MS: m / z calcd.for[C 25 H 32 N2O5S+H] + 473.2, found 473.3.

[0120] Example 30: Synthesis of Compound 30

[0121]

[0122] According to the synthesis method of compound 28, a yellow solid (433 mg, 56%) was obtained by column chromatography (PE / EA=2 / 1, v / v). 1 H NMR (600MHz, CDCl3) δ8.08 (d, J=8.5Hz, 2H), 7.37 (d, J=8.8Hz, 1H), 7.20 (s, 1H), 6.89 (d, J=8.8Hz, 1H), 6.76 (d, J=8.5Hz , 2H), 4.18 (t, J=4.5Hz, 2H), 4.04 (s, 2H), 3.90 (t, J=4.5Hz, 2H), 3.77 (d, J=7.7Hz, 4H), 3.06 (s, 6H), 1.47 (s, 9H).MS: m / z calcd.for[C 25 H 32 N2O6+H] + 457.2, found 457.3.

[0123] Example 31: Synthesis of Compound 31

[0124]

[0125] Compound 28 (229 mg, 0.5 mmol) was dissolved in DCM (8 mL), TFA (4 mL) was added dropwise, and the reaction was stirred at room temperature for 3 h. The solvent was removed under reduced pressure and the mixture was washed with ether to obtain a red solid (169 mg, 84%). 1 H NMR (600MHz, CDCl3) δ7.88 (d, J=9.0Hz, 1H), 7.80 (d, J=8.8Hz, 2H), 7.29 (d, J=2.4Hz, 1H), 7.14 (dd, J=9.0, 2.5Hz, 1H) , 6.61 (d, J=8.8Hz, 2H), 4.21 (dd, J=8.1, 2.9Hz, 2H), 4.15 (s, 2H), 3.93-3.91 (m, 2H), 3.77 (s, 4H), 2.90 (s, 3H).MS: m / z calcd.for[C 20 H 22 N2O5S+H] + 403.1, found 403.3.

[0126] Example 32: Synthesis of Compound 32

[0127]

[0128] According to the synthesis method of compound 31, a red solid (201 mg, 88%) was obtained. 1H NMR (600MHz, CDCl3) δ7.88 (t, J=9.1Hz, 3H), 7.30 (s, 1H), 7.16 (d, J=8.9Hz, 1H), 6.75 (d, J=8. 4Hz, 2H), 4.22-4.19(m, 2H), 4.17(s, 2H), 3.93-3.90(m, 2H), 3.78(s, 4H), 3.12(s, 6H).MS: m / z calcd.for[C 21 H 24 N2O5S+H] + 417.1, found 417.3.

[0129] Example 33: Synthesis of Compound 33

[0130]

[0131] According to the synthesis method of compound 31, a yellow solid (343 mg, 90%) was obtained. 1 H NMR (400MHz, CDCl3) δ8.09 (d, J=8.5Hz, 2H), 7.38 (d, J=8.8Hz, 2H), 6.90 (d, J=10.1Hz, 1H), 6.77 (d, J=8.6Hz, 2 H), 4.26-4.22 (m, 2H), 4.20 (s, 2H), 3.92 (d, J=4.1Hz, 2H), 3.83-3.75 (m, 4H), 3.07 (s, 6H).MS: m / zcalcd.for[C 21 H 24 N2O6+H] + 401.2, found 401.3.

[0132] Example 34: Synthesis of Compound 34

[0133]

[0134] According to the synthesis method of compound 20, a yellow solid (311 mg, 53%) was obtained by column chromatography (PE / EA=1 / 4, v / v). 1H NMR (600MHz, CDCl3) δ7.83 (t, J=8.6Hz, 3H), 7.33-7.29 (m, 6H), 7.02 (dd, J=8.9, 2.5Hz, 1H) , 6.61 (d, J=8.7Hz, 2H), 5.06 (s, 2H), 4.50 (m, 5.1Hz, 1H), 4.25-4.20 (m, 1H), 4.16 (dd, J=14. 3, 4.9Hz, 1H), 4.01 (s, 2H), 3.91-3.86 (m, 2H), 3.75-3.72 (m, 2H), 3.71-3.63 (m, 2H), 3.11 ( m, 2H), 2.86 (s, 3H), 1.79-1.72 (m, 1H), 1.61 (m, 1H), 1.42 (s, 9H), 1.29-1.24 (m, 4H).MS: m / z calcd.for[C 38 H 48 N4O8S+H] + 721.3, found 721.3.

[0135] Example 35: Synthesis of Compound 35

[0136]

[0137] According to the synthesis method of compound 20, a yellow solid (251 mg, 68%) was obtained by column chromatography (PE / EA=1 / 4, v / v). 1H NMR (600MHz, CDCl3) δ7.91 (d, J=8.5Hz, 2H), 7.86 (t, J=7.6Hz, 1H), 7.34 (d, J=3.6Hz, 3H), 7.32 (d, J=2.5Hz, 1H), 7.31-7.28 (m, 2H), 7 .04 (dd, J=8.9, 2.5Hz, 1H), 6.73 (d, J=8.6Hz, 2H), 5.07 (s, 2H), 4.52 (m, 1H), 4.27-4.23 (m, 1H), 4.20-4.16 (m, 1H), 4.02 (d, J=2.7Hz, 2 H), 3.91 (m, 2H), 3.75 (d, J=3.5Hz, 2H), 3.67 (q, J=6.5Hz, 2H), 3.16 (dd, J=13.3, 6.7Hz, 1H), 3.09 (dd, J=13.3, 6.5Hz, 1H), 3.05 (s, 6H) , 1.80-1.73 (m, 1H), 1.66-1.59 (m, 1H), 1.45 (d, J=8.2Hz, 2H), 1.43 (s, 9H), 1.38-1.32 (m, 1H), 1.31-1.26 (m, 1H).MS: m / zcalcd.for[C 39 H 50 N4O8S+H] + 735.3, found 735.3.

[0138] Example 36: Synthesis of Compound 36

[0139]

[0140] According to the synthesis method of compound 20, a yellow solid (481 mg, 76%) was obtained by column chromatography (PE / EA=1 / 4, v / v). 1 H NMR (400MHz, CDCl3) δ8.13 (d, J=8.5Hz, 2H), 7.41 (q, J=10.3Hz, 7H), 6.94 (d, J=8.8Hz, 1H) , 6.79 (d, J=8.5Hz, 2H), 5.14 (s, 2H), 4.58 (q, J=7.6Hz, 1H), 4.32-4.19 (m, 2H), 4.09 (s, 2H ), 3.96 (s, 2H), 3.85-3.72 (m, 4H), 3.21 (d, J = 11.9Hz, 2H), 3.10 (s, 6H), 1.84 (d, J = 8.9Hz, 1H), 1.72 (d, J=7.7Hz, 1H), 1.60-1.52 (m, 2H), 1.51 (s, 9H), 1.42 (d, J=13.6Hz, 2H).MS: m / z calcd.for[C39 H 50 N4O9+H] + 719.4, found 719.3.

[0141] Example 37: Synthesis of Compound 37

[0142]

[0143] According to the synthesis method of compound 31, a red solid (263 mg, 92%) was obtained. 1 H NMR (400MHz, CDCl3) δ7.76 (dd, J=21.5, 8.4Hz, 3H), 7.62 (s, 1H), 7.48 (d, J=7.7Hz, 1H), 7.28 (s, 3H), 7.17 (s, 1H), 6.95 (d, J=10.9Hz, 1H), 6.53 (d, J=8.1Hz, 2H), 5.03 (s, 2H), 4.59 (t, J=7.2Hz, 1H), 4.01 (m, 4H), 3.78 (s, 2H), 3.66 (s, 4H), 3.06 (m, 2H) , 2.77 (s, 3H), 1.83 (t, J=12.4Hz, 1H), 1.73-1.61 (m, 1H), 1.45-1.30 (m, 4H).MS: m / z calcd.for[C 34 H 40 N4O8S+H] + 665.3, found665.3.

[0144] Example 38: Synthesis of Compound 38

[0145]

[0146] According to the synthesis method of compound 31, a red solid (206 mg, 89%) was obtained. 1H NMR (600MHz, CDCl3) δ7.94 (d, J=8.9Hz, 1H), 7.89 (d, J=8.7Hz, 2H), 7.39 (d, J=8.1Hz, 1H), 7 .31 (d, J=13.2Hz, 5H), 7.13 (d, J=9.0Hz, 1H), 6.72 (d, J=8.7Hz, 2H), 5.06 (s, 2H), 4.51 (d, J =7.5Hz, 1H), 4.24-4.17(m, 2H), 4.05(t, J=17.8Hz, 2H), 3.86(t, J=4.8Hz, 2H), 3.74-3.65( m, 4H), 3.09 (s, 8H), 1.81 (m, 1H), 1.65 (m, 1H), 1.48-1.41 (m, 2H), 1.35-1.27 (m, 2H).MS: m / z calcd.for[C 35 H 42 N4O8S+H] + 679.3.found 679.3.

[0147] Example 39: Synthesis of Compound 39

[0148]

[0149] According to the synthesis method of compound 31, a yellow solid (416 mg, 94%) was obtained. 1 H NMR (400MHz, CDCl3) δ8.19 (d, J=8.8Hz, 2H), 7.83 (d, J=7.5Hz, 1H), 7.53 (s, 1H), 7.47 (d, J=8.5H z, 1H), 7.34 (s, 5H), 7.01 (d, J=9.7Hz, 1H), 6.81 (d, J=8.9Hz, 2H), 5.07 (s, 2H), 4.64 (q, J=6.0Hz, 1H), 4.48 (t, J=8.8Hz, 1H), 4.25-4.16 (m, 1H), 4.10 (m, 1H), 4.02-3.85 (m, 3H), 3.68 (m, 4H), 3.1 7(s, 7H), 3.00-2.89(m, 1H), 1.84-1.66(m, 2H), 1.46-1.34(m, 3H), 1.14(d, J=7.3Hz, 1H).MS: m / z calcd.for[C 35 H 42 N4O9+H] + 663.3, found 663.4.

[0150] Example 40: Synthesis of Compound 40

[0151]

[0152] Compound 37 (126 mg, 0.2 mmol), compound 26 (130 mg, 0.3 mmol), HOBT (39 mg, 0.3 mmol), EDCI (56 mg, 0.3 mmol), DIPEA (37 mg, 0.3 mmol), and a catalytic amount of DMAP were added to 10 mL of DCM and stirred at room temperature overnight. The mixture was concentrated under reduced pressure, dissolved in EA (50 mL), washed with H2O, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by column chromatography (DCM / MeOH = 9 / 1, v / v) to obtain a yellow oil (146 mg, 70%). 1 H NMR (400MHz, CDCl3) δ8.33 (d, J=5.8Hz, 1H), 7.83 (t, J=8.2Hz, 3H), 7.71 (d, J=10.5Hz, 1H), 7.49 (d, J=8.3Hz, 1H), 7.35-7.28 (m, 6H ), 7.04 (dd, J=8.8, 2.4Hz, 1H), 6.62 (d, J=8.4Hz, 2H), 5.16-5.01 (m, 3H), 4.65 (q, J=6.9Hz, 1H), 4.33-4.26 (m, 1H), 4.23-4.09 (m, 9H ), 4.04 (d, J = 13.4Hz, 2H), 3.98 (s, 2H), 3.90 (d, J = 4.5Hz, 2H), 3.73 (d, J = 4.6Hz, 2H), 3.67 (d, J = 5.0Hz, 2H), 3.61 (d, J = 4.6Hz, 2H), 3.55 (d, J=4.8Hz, 2H), 3.50 (s, 4H), 3.09 (m, 2H), 2.88 (s, 3H), 1.81-1.61 (m, 2H), 1.31 (d, J=7.1Hz, 6H), 1.28-1.24 (m, 10H). MS: m / z calcd.for[C 49 H 72 N6O 16 P2S+H] + 1095.4, found1095.4.

[0153] Example 41: Synthesis of Compound 41

[0154]

[0155] According to the synthesis method of compound 40, a yellow oil (276 mg, 66%) was obtained by column chromatography (DCM / MeOH=9 / 1, v / v). 1H NMR (600MHz, CDCl3) δ8.36 (s, 1H), 7.97 (s, 3H), 7.71 (d, J = 10.4Hz, 1H), 7.48 (d, J = 8.3Hz, 1H), 7.34-7.27 (m, 6H), 7.08 (d, J = 8.1Hz, 1H), 6 .76 (d, J=7.7Hz, 2H), 5.13-5.02 (m, 3H), 4.68-4.62 (m, 1H), 4.30 (m, 1H), 4.21 (m, 5H), 4.14-4.09 (m, 4H), 4.04 (m, 2H), 3.98 (d, J=6.9Hz, 2H ), 3.91 (d, J = 4.7Hz, 2H), 3.74 (t, J = 4.5Hz, 2H), 3.67 (dt, J = 10.2, 4.5Hz, 2H), 3.61 (dd, J = 9.1, 4.6Hz, 2H), 3.56 (m, 2H), 3.50 (q, J = 6.1Hz, 4H), 3.15-3.09 (m, 1H), 3.06 (s, 7H), 1.75 (q, J=6.8Hz, 1H), 1.70-1.63 (m, 1H), 1.50-1.37 (m, 2H), 1.32 (m, 4H), 1.30-1.23 (m, 10H). MS: m / z calcd.for[C 50 H 74 N6O 16 P2S+H] + 1109.4, found1109.4.

[0156] Example 42: Synthesis of Compound 42

[0157]

[0158] According to the synthesis method of compound 40, a colorless oil (197 mg, 50%) was obtained by column chromatography (DCM / MeOH=9 / 1, v / v). 1H NMR (600MHz, CDCl3) δ8.31 (t, J=5.7Hz, 1H), 8.00 (d, J=8.9Hz, 2H), 7.69 (d, J=9.0Hz, 1H), 7.46 (d, J=8.1Hz, 1H), 7.32-7.22 (m, 6H), 7.15 (d, J=2.5Hz, 1H), 6.83 (dd, J=8.7, 2.5Hz, 1H), 6.68 (d, J=8.5Hz, 2H), 5.10-4.97 (m, 3H), 4.61 (q, J=7.0Hz, 1H), 4.23-4.05 (m, 10 H), 3.96 (m, 4H), 3.88-3.81 (m, 2H), 3.71-3.66 (m, 2H), 3.62 (dd, J=9.7, 4.6Hz, 2H), 3.59-3.55 (m, 2H), 3.53 (d, J=4.3Hz, 2H), 3.48-3 .43 (m, 4H), 3.08 (q, J=6.0Hz, 1H), 2.99 (s, 7H), 1.76-1.68 (m, 1H), 1.68-1.59 (m, 1H), 1.47-1.34 (m, 2H), 1.30-1.21 (m, 14H).MS: m / z calcd.for[C 50 H 74 N6O 17 P2+H] + 1093.5, found 1093.5.

[0159] Example 43: Synthesis of Compound 43

[0160]

[0161] According to the synthesis method of compound 18, a yellow solid (114 mg, 89%) was obtained and used directly in the next reaction without purification.

[0162] Example 44: Synthesis of Compound 44

[0163]

[0164] According to the synthesis method of compound 18, a yellow solid (219 mg, 90%) was obtained and used directly in the next reaction without purification.

[0165] Example 45: Synthesis of Compound 45

[0166]

[0167] According to the synthesis method of compound 18, a yellow solid (149 mg, 86%) was obtained and used directly in the next reaction without purification.

[0168] Example 46: Synthesis of Compound 46

[0169]

[0170] According to the synthesis method of compound 40, a yellow solid (80 mg, 66%) was obtained by column chromatography (DCM / MeOH=6 / 1, v / v). 1 H NMR (600MHz, CDCl3) δ8.50 (t, J=5.6Hz, 1H), 8.14 (d, J=5.8Hz, 2H), 7.82 (m, 3H), 7.67 (d, J=8.2Hz, 1H), 7.45 (d, J=10.3Hz, 1H), 7.35 (d, J=2.6Hz, 1 H), 7.04 (dd, J=8.9, 2.6Hz, 1H), 6.63 (d, J=8.7Hz, 2H), 5.05 (td, J=21.9, 10.4Hz, 1H), 4.59-4.54(m, 1H), 4.33(m, 1H), 4.24-4.16(m, 11H), 4.09-4 .00 (m, 4H), 3.91 (d, J = 5.3Hz, 2H), 3.80-3.77 (m, 2H), 3.73 (dd, J = 11.0, 3 .8Hz, 2H), 3.68 (t, J=4.4Hz, 2H), 3.63-3.58 (m, 4H), 3.48 (q, J=5.9Hz, 2H ), 3.15(s, 8H), 2.89(s, 3H), 2.83-2.00(m, 16H), 1.87(m, 1H), 1.68(m, 1H ), 1.56-1.46 (m, 4H), 1.42 (d, J=10.8Hz, 27H), 1.34-1.30 (m, 12H).MS: m / z calcd.for[C 69 H 116 N 10 O 21 P2S+Na] + 1537.8, found 1537.6.

[0171] Example 47: Synthesis of Compound 47

[0172]

[0173] According to the synthesis method of compound 40, a yellow solid (103 mg, 48%) was obtained by column chromatography (DCM / MeOH=6 / 1, v / v). 1H NMR (400MHz, CDCl3) δ8.82 (t, J=5.8Hz, 1H), 8.19 (s, 1H), 7.86 (d, J=8.9Hz, 2H), 7.80 (d, J=8.9Hz, 1H), 7.70 (d, J=8.1Hz, 1H), 7.41 (d, J=10.3Hz, 1H), 7. 34 (d, J=2.6Hz, 1H), 7.03 (dd, J=8.9, 2.5Hz, 1H), 6.71 (d, J=9.0Hz, 2H), 5.0 3(td, J=21.9, 10.4Hz, 1H), 4.56 (td, J=8.3, 4.8Hz, 1H), 4.23-4.13 (m, 10H), 4.05 (m, 4H), 3.89 (dd, J=5.7, 3.9Hz, 2H), 3.79-3.75 (m, 2H), 3.73 (t, J=4.1 Hz, 2H), 3.68-3.65 (m, 2H), 3.60 (q, J=5.6Hz, 4H), 3.48-3.43 (m, 4H), 3.38-3 .08(m, 8H), 3.02(s, 6H), 2.27(m, 16H), 1.90(d, J=8.7Hz, 1H), 1.75-1.65(m , 1H), 1.52 (dt, J=13.1, 6.4Hz, 4H), 1.45-1.38 (m, 39H).MS: m / zcalcd.for[C 70 H 118 N 10 O 21 P2S+H] + 1551.8, found 1551.5.

[0174] Example 48: Synthesis of Compound 48

[0175]

[0176] According to the synthesis method of compound 40, a yellow solid (152 mg, 67%) was obtained by column chromatography (DCM / MeOH=6 / 1, v / v). 1H NMR (600MHz, CDC13) δ 8.57 (s, 1H), 8.15 (s, 1H), 7.95 (d, J = 7.9Hz, 2H), 7.58 (d, J = 7.9Hz, 1H), 7.37 (d, J = 10.0Hz, 1H), 7.28 (d, J = 9. 0Hz, 1H), 7.09 (s, 1H), 6.79 (d, J=8.7Hz, 1H), 6.64 (d, J=8.1Hz, 2H), 4.96 (td, J=21.8, 10.3Hz, 1H), 4.50 (d, J=12.3Hz, 1H), 4.16-4 .04 (m, 10H), 3.96 (m, 4H), 3.81 (s, 2H), 3.66 (m, 4H), 3.59 (d, J=7.1Hz, 2H), 3.55-3.49 (m, 4H), 3.44-3.01 (m, 10H), 2.96 (s, 6H), 2. 80-1.99 (m, 18H), 1.80 (s, 1H), 1.61 (s, 1H), 1.47-1.41 (m, 2H), 1.34 (d, J=9.3Hz, 35H), 1.22 (d, J=7.1Hz, 6H).MS: m / zcalcd.for[C 70 H 118 N 10 O 20 P2+H] + 1535.8, found 1535.6.

[0177] Example 49: Synthesis of Compound 49

[0178]

[0179] According to the synthesis method of compound 24, a yellow solid (54 mg, 83%) was obtained. 1H NMR (400MHz, DMSO-d6) δ8.53 (s, 1H), 8.37 (s, 1H), 7.77 (t, J=9.8Hz, 3H), 7.73-7.48 (m, 3H), 7.2 6 (d, J=10.3Hz, 1H), 7.06 (d, J=8.9Hz, 1H), 6.64 (d, J=8.4Hz, 2H), 4.47 (m, 1H), 4.33 (m, 1H), 4.19 (m, 2H), 4.08 (m, 2H), 3.92 (m, 8H), 3.80 (m, 2H), 3.63 (m, 10H), 3.51 (m, 2H), 3.41 (m, 4H), 3.29-3. 23(m, 4H), 3.11(s, 12H), 2.75(s, 3H), 1.64(m, 1H), 1.53(m, 1H), 1.40(m, 2H), 1.24-1.21(m, 2H). 13 C NMR (101MHz, DMSO-d6) δ172.33, 171.87, 169.38, 168.86, 166.26, 156.40, 152.70, 148.88, 135.59, 128.83, 122.80, 120.74, 116.01, 112.04, 107 .50, 106.20, 70.78, 70.24, 69.77, 69.47, 68.24, 55.23, 54.43, 53.11, 5 2.22, 51.10, 48.87, 48.51, 39.10, 32.86, 29.86, 28.86, 23.09.HRMS: m / z calcd for[C 49 H 76 N 10 O 21 P2S-H] - 1233.4304.found1233.4273.

[0180] Example 50: Synthesis of Compound 50

[0181]

[0182] According to the synthesis method of compound 24, a yellow solid (72 mg, 85%) was obtained. 1H NMR (400MHz, DMSO-d6) δ8.54 (s, 1H), 8.42 (s, 1H), 7.82 (m, 3H), 7.73-7.58 (m, 2H), 7.25 (d, J=10 .1Hz, 1H), 7.08 (dd, J=8.9, 2.7Hz, 1H), 6.81 (d, J=9.1Hz, 2H), 4.53-4.39 (m, 1H), 4.39-4.29 (m, 1 H), 4.19 (d, J=5.9Hz, 2H), 3.94 (m, 8H), 3.81 (m, 2H), 3.68-3.56 (m, 10H), 3.51 (m, 2H), 3.43 (m, 2 H), 3.34-3.19(m, 10H), 3.09(m, 10H), 3.01(s, 6H), 1.59(m, 2H), 1.40(m, 2H), 1.27-1.20(m, 2H). 13 C NMR (101MHz, DMSO-d6) δ171.63, 171.35, 168.80, 168.32, 165.43, 155.96, 151.92, 148.35, 135.17, 128.09, 122.39, 120.43, 118.63, 115.57, 111.85, 105.67, 70.23, 69.70, 69.21, 68.93, 67.72, 54.73, 53.87, 52. 86, 51.74, 50.47, 48.54, 48.30, 38.57, 32.36, 28.31, 22.53.HRMS: m / z calcd for[C 50 H 78 N 10 O 21 P2S-H]-1247.4466.found 1247.4425.

[0183] Example 51: Synthesis of Compound 51

[0184]

[0185] According to the synthesis method of compound 24, a yellow solid (65 mg, 80%) was obtained. 1H NMR (400MHz, DMSO-d6) δ8.54 (s, 1H), 8.35 (s, 1H), 7.96 (d, J = 8.6Hz, 2H), 7.66 (d, J = 8.4Hz, 1H), 7.57 (d, J = 8.8Hz, 1H), 7 .30-7.23 (m, 2H), 6.91 (dd, J=8.8, 2.5Hz, 1H), 6.84 (d, J=8.7Hz, 2H), 4.46 (td, J=20.6, 10.3Hz, 1H), 4.33 (q, J=7.4Hz, 1 H), 4.19-4.14 (m, 2H), 4.10 (d, J=12.4Hz, 2H), 3.93 (d, J=15.6Hz, 6H), 3.82-3.77 (m, 2H), 3.70-3.47 (m, 14H), 3.43 (t, J =5.7Hz, 8H), 3.31-3.21(m, 4H), 3.11(s, 8H), 3.03(s, 6H), 1.71-1.48(m, 2H), 1.40(d, J=7.6Hz, 2H), 1.28-1.19(m, 2H). 13 C NMR (101MHz, DMSO-d6) δ172.32, 171.88, 169.40, 169.29, 168.86, 164.70, 156.54, 152.90, 145.08, 143.50, 129.05, 113.47, 112.88, 112.29, 110.98, 103.83, 70.78, 70.25, 69.79, 69.53, 68.35, 55.24, 54.49, 53. 12, 52.24, 51.12, 48.91, 46.40, 39.10, 32.85, 28.86, 23.10.HRMS: m / z calcd for[C 50 H 78 N 10 O 22 P2-H] - 1231.4695.found 1231.4664.

[0186] Example 52: Radiolabeling

[0187] 1. Experimental steps:

[0188]

[0189] The germanium gallium generator was eluted with 3 mL of HCl (0.1 M) to obtain [ 68Ga]GaCl3 solution was added to a mixed solution of precursor 49 or 50 (20 μg) and NaOAc (390 μL, 1.0 M), the reaction bottle was sealed and placed at 90°C for reaction for 10 minutes.

[0190] 2. Experimental Results

[0191] [ 68 Ga]Ga-49 and [ 68 After HPLC separation and purification, the radiochemical purity of Ga]Ga-50 was greater than 95%, allowing direct use in subsequent experiments. The chromatographic column was a Venusil MP C18 reverse phase column (10 μm, 10 mm × 250 mm), with a mobile phase flow rate of 4 mL / min. The labeling rates are shown in Table 1.

[0192] Table 1 Labeling rate of labeled compounds (mobile phase: acetonitrile / H2O (0.2% H3PO4) = 28 / 72, v / v)

[0193]

[0194] Example 53: Determination of affinity of compounds to ATTR

[0195] 1. Experimental steps:

[0196] ATTR aggregates (final concentration 288 nM), fluorescent tracer (final concentration 500 nM), gradient concentrations of test compounds (final concentration 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -8.5 , 10 -9 , 10 -10 M) and PBS (1×), and the fluorescence signal was detected using an Infinite M200 pro microplate reader (excitation / emission wavelengths were 520 / 610 nm). Nonspecific fluorescence was determined by setting up a parallel experimental group without adding ATTR aggregates. GraphPad Prism 9.0 was used for analysis, and K was calculated according to the Cheng-Prusoff equation. i Value (K i =IC 50 / (1+[L] / Kd)).

[0197] 2. Experimental results:

[0198] As shown in Table 2, compounds 15, 16, 49 and 51 of the present invention have moderate affinity for ATTR, while compound 50 has a higher affinity for ATTR (K i =4.9±2.5nM).

[0199] Table 2 Affinity data of compounds with ATTR

[0200]

[0201] Example 54: Ca 2+ Binding capacity assay

[0202] 1. Experimental steps:

[0203] (1) Prepare calcium standard solutions with gradient concentrations (0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0 mM);

[0204] (2) Calcium standard solutions of different concentrations and the detection working solution were mixed in a 96-well plate, incubated in the dark at room temperature for 10 min, and the absorbance at 575 nm was measured by a microplate reader to prepare a standard curve;

[0205] (3) The calcium standard solution, the test solution, and the detection working solution were mixed in a 96-well plate and incubated in the dark at room temperature for 10 min. The absorbance at 575 nm was measured using a microplate reader. The calcium content was calculated based on the standard curve, and the calcium binding rate was also calculated.

[0206] 2. Experimental results:

[0207] like Figure 4 As shown, compound 50 and Ca 2+ The compounds exhibited strong chelation effects, with chelation rates of 51.4%, 86.1%, and 98.6%, respectively, comparable to the positive control EDTA (67.6%, 95.8%, and 97.2%). The excellent calcium chelation ability further demonstrates that compound 50 has great potential in ATTR imaging.

[0208] Example 55: Autoradiography Experiment

[0209] A certain concentration of labeled product (10% ethanol solution) was incubated with sections of ATTR patients, AL patients, Aβ patients and healthy volunteers at room temperature for a certain period of time. After incubation, the sections were exposed through a phosphor screen and the images were analyzed using a storage phosphor screen system.

[0210] 1. Experimental steps:

[0211] (1) Pretreatment of sections (dewaxing and rinsing);

[0212] (2) Cover the slices with 20 μCi / mL of 68 1 mL of Ga-labeled compound solution was incubated at room temperature for 1 h;

[0213] (3) Rinse with 20% ethanol for 1 minute;

[0214] (4) After drying, place it under a phosphor screen and expose it for 1 hour. Then use a storage phosphor screen system to analyze the image.

[0215] 2. Experimental results:

[0216] The experimental results are as follows Figure 5 As shown, [ 18 A strong specific signal of F]16 was observed in ATTR slices, a relatively weak radioactive signal concentration was found in AL and Aβ slices, and no obvious signal was detected in healthy myocardial slices. 68 Ga]Ga-49 and [ 68 Ga]Ga-50 showed significant radioactive signal accumulation in ATTR slices, while no significant signal was detected in AL, Aβ, and healthy myocardial slices. This fully demonstrates that the compound of the present invention, after being labeled with radionuclides, can be used as an ATTR imaging agent and has potential application prospects in clinical diagnosis.

[0217] Example 56: Micro-PET / CT imaging experiment in SD rats

[0218] 1. Experimental steps:

[0219] Micro-PET / CT imaging experiments in normal SD rats will 68 Ga]Ga-49 or [ 68 After the Ga]Ga-50 injection was injected into rats (n=2-3, 6 weeks old, male) through the tail vein, a 0-60 min PET scan was immediately started. During the scan, anesthesia was performed by continuous inhalation of a mixture of isoflurane and air at a gas flow rate of 1-2 L / min. After the PET data acquisition was completed, the rat CT data acquisition was completed to obtain anatomical information. The PET data was reconstructed in 3D-OSEM mode, and the dynamic data was framed in the form of 60s×15, 300s×5, and 600s×2. The reconstructed data was imported into PMOD 4.1 software to complete the quantitative analysis of the data and export the imaging result images. The animal experiments complied with the relevant regulations of the Animal Ethics Committee of Beijing Normal University.

[0220] 2. Experimental results:

[0221] PET / CT results such as Figure 6 and Figure 7 , [ 68 Ga]Ga-50 has a rapid and efficient uptake in the myocardium, reaching a peak uptake in the myocardium (SUV = 4.8) 1 minute after administration, and then rapidly cleared from the healthy myocardium. In addition, non-target organs such as the liver and lungs maintain low uptake throughout the imaging period. 2+ Chelation, the radioactivity in the bones increased over time, and the spine and major joints could be clearly observed at 60-70 minutes.68 The PET data of Ga]Ga-49 and [ 68 This indicates that the molecular skeleton generally has good biological properties and has potential application prospects in the clinical diagnosis of ATTR.

Claims

1. A compound of formula (I), characterized in that in X1 is selected from S or O; X2 is selected from N or CH; When R1 is fluorinated oligoethylene glycol, R2=R3, and is selected from the group consisting of: -C 1-4 Alkyl-carboxyl, -C 1-4 Alkyl-bisphosphonic acid group; When R1 is When R2 and R3 are each independently selected from -CH3 or -H, wherein R4 is selected from the group consisting of: -C 1-10 Alkyl, -oligoethylene glycol; R5 is selected from the group consisting of: -C 1-10 Alkyl-chelate group, -oligoethylene glycol-chelate group; R6 is selected from the group consisting of: -C 1-10 Alkyl-bisphosphonic acid group, -oligoethylene glycol-bisphosphonic acid group; The oligoethylene glycol is a oligoethylene glycol composed of 1-5 ethylene glycols; the fluorinated oligoethylene glycol means that the terminal hydroxyl group of the oligoethylene glycol is substituted by fluorine; wherein the bisphosphonic acid group is The chelating group is a chelating radionuclide 68 Ga, 64 Cu, Al 18 F. 99m A chelating group for Tc or the corresponding stable isotope.

2. The compound according to claim 1, characterized in that When R1 is fluorinated oligoethylene glycol, R2=R3, and is selected from the group consisting of: -C 1-4 Alkyl-carboxyl, -C 1-4 Alkyl-bisphosphonic acid group; preferably, R1 is -(CH2CH2O)2CH2CH2F, and R2=R3, selected from the group consisting of: -CH2CH2COOH, -CH2COOH, 3. The compound according to claim 1, characterized in that When R1 is When R2 and R3 are each independently selected from -CH3 or -H, wherein R4 is selected from the group consisting of: -C 1-10 Alkyl, -oligoethylene glycol; R5 is selected from the group consisting of: -C 1-10 Alkyl-chelate group, -oligoethylene glycol-chelate group; R6 is selected from the group consisting of: -C1- 10 Alkyl-bisphosphonic acid group, -oligoethylene glycol-bisphosphonic acid group; preferably, R4 is -(CH2CH2O)2CH2CONH-; R5 is selected from And R d for Where M is 68 Ga 3+ 、 67 Ga 3+ or its corresponding stable isotope.

4. The compound according to any one of claims 1 to 3, characterized in that The compound is selected from the group consisting of:

5. The radiolabeled compound of any one of claims 1 to 4, wherein The radiolabeled compound is prepared by the method of any one of claims 1 to 4. 18 F, 68 Ga or 67 Ga labeled.

6. A pharmaceutical composition, characterized in that Comprising the compound according to any one of claims 1 to 4, or the radiolabeled compound according to claim 5; and a pharmaceutically acceptable carrier.

7. A diagnostic or detection reagent, characterized in that Comprising the compound according to any one of claims 1 to 4, or the radiolabeled compound according to claim 5; and a pharmaceutically acceptable carrier.

8. The diagnostic or detection reagent according to claim 7, characterized in that For the diagnosis or detection of transthyretin amyloidosis.

Images