Aldehyde hydrazone light cage compound as well as preparation method and application thereof

Abstract

The invention provides an aldehyde hydrazone light cage compound which can absorb near ultraviolet light to visible light, and can be subjected to light degradation in a specific wavelength range to release corresponding aldehyde and phenyl compounds. The structure of the aldehyde hydrazone compound can be used as a universal strategy for designing a light release prodrug with high space-time controllability; the material can be degraded by receiving light in vivo and in vitro; and the medicine can also be used as an auxiliary medicine for photodynamic therapy.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to aldehyde hydrazone photocage compounds, their preparation methods, and applications. Background Technology

[0002] Photocage compounds are a novel type of light-controlled release compound. By linking a target drug to a photoreleasable group, the drug's activity is "cageed," temporarily losing its biological, chemical, and optical activity. Upon light irradiation, the drug is released and activated in a specific spatiotemporal context, thus exerting its active compound effect. Photocages have become a current research hotspot due to their unique advantages, including non-invasive control, high spatiotemporal precision, and high biocompatibility.

[0003] Early research on photocage drugs focused primarily on the selection and synthesis of photosensitive groups to identify those capable of efficient and rapid light response. As research progressed, scientists gradually began to focus on the behavior of photocage drugs in vivo, including drug distribution, metabolism, and toxicity. Currently, limitations remain, including a limited range of photocage structural types, short degradation wavelengths, low tissue penetration, and the inability to release aldehyde compounds, which restrict the development of photocage prodrugs. Summary of the Invention

[0004] To address the problems in the prior art, this invention provides a novel aldehyde hydrazone photocage compound. This type of compound has strong photocage degradation ability, can release aldehyde compounds, and has strong tissue penetration ability, thus possessing significant development value.

[0005] The present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides an aldehyde hydrazone photocage compound.

[0007] Aldehyde hydrazone photocage compounds have the following structural formula (I):

[0008]

[0009] in

[0010] Z is selected from C or N;

[0011] W is selected from C or N;

[0012] R1 is 4-(1,2,2-triphenylvinyl)phenyl, anthracene, 3-coumarinyl, 4-diphenylaminophenyl (TPA), 1-pyrene, 3-(4-(dimethylamino)phenyl)propenyl, pyrene, cyaninyl, strophanthidinyl, or rhodamine 6G;

[0013] R2 is selected from H, halogen, nitro, C1-C6 alkyl, COOR3, OR4, SO2R5 or cyano; R3 is H or C1-C6 alkyl; R4 is H, benzyl, C1-C6 alkyl or halogen-substituted C1-C6 alkyl; R5 is H, C1-C6 alkyl or amino.

[0014] According to one embodiment of the present invention, R1 is selected from the following groups:

[0015]

[0016] According to one embodiment of the present invention, in the structural formula shown in formula (I), the number of R2 substitutions on the aromatic ring is 1-5, and the R2 substitution position on the aromatic ring is adjacent, meta or para, or adjacent, meta or para substitution individually or simultaneously. When one R2 group is substituted on the aromatic ring in the structural formula shown in formula (I), such as 2-R2, 3-R2, 4-R2; when two R2 groups are substituted on the aromatic ring, such as 2,3-(R2)2, 2,4-(R2)2, 2,5-(R2)2, 3,4-(R2)2, 3,5-(R2)2; when three R2 groups are substituted on the aromatic ring, such as 2,3,4-(R2)3, 2,3,5-(R2)3, 2,4,6-(R2)3; when four R2 groups are substituted on the aromatic ring, such as 2,3,4,5-(R2)4, 2,3,4,6-(R2)4; when five R2 groups are substituted on the aromatic ring, such as 2,3,4,5,6-(R2)5, Z, W, and R2 are the same as above.

[0017] According to one embodiment of the present invention, Z is C, and W is selected from C or N.

[0018] According to one embodiment of the present invention, Z is N, and W is selected from C or N.

[0019] According to one embodiment of the present invention, the above-mentioned aldehyde hydrazone photocage compound has the following structural formula:

[0020]

[0021] R1 and R2 are the same as above.

[0022] According to one embodiment of the present invention, the aldehyde hydrazone photocage compound of the present invention is selected from the following general formula compounds (YB, CYHR, BW, CY5Q, TP, FY, LG, SA, PC, CM, LZ):

[0023]

[0024]

[0025] Z, W, and R2 are the same as above, while A is an anion.

[0026] According to one embodiment of the present invention, the aforementioned anion is an inorganic anion or an organic anion. Inorganic anions include chloride ions, bromide ions, iodide ions, sulfate ions, sulfite ions, nitrate ions, nitrite ions, phosphate ions, hydrogen phosphate ions, etc. Organic anions include acetate ions, propionate ions, cinnamate ions, benzosulfonate ions, citrate ions, lactate ions, gluconate ions, etc.

[0027] As used in this article, the term "halogen" refers to fluorine, chlorine, bromine, or iodine. As used in this article, the term "C" refers to... 1-6 "Alkyl" refers to a saturated straight-chain or branched hydrocarbon group or cycloalkyl group having 1-6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, etc.

[0028] The term “halogen-substituted C1-6 alkyl” as used in this article refers to one or more halogen-substituted saturated straight-chain or branched hydrocarbon groups having 1-6 carbon atoms, such as difluoromethyl, trifluoromethyl, etc.

[0029] According to one embodiment of the present invention, the R2 is selected from H, fluorine, chlorine, bromine, iodine, nitro, cyano, methyl, ethyl, propyl, isopropyl, benzyloxy, -COOH, -COOCH3, -COOCH2CH3, -OH, -OCH3, -OCH2CH3, -OCF3, -SO2CH3 or -SO2NH2.

[0030] According to one embodiment of the present invention, the aldehyde hydrazone photocage compound of the present invention preferably contains the following compounds:

[0031]

[0032]

[0033]

[0034]

[0035] Secondly, the present invention provides a method for preparing the above-mentioned aldehyde hydrazone photocage compound.

[0036] The above-mentioned aldehyde hydrazone photocage compound is prepared by condensation reaction of formula (III) and formula (IV) to obtain compound (I), and the reaction formula is as follows:

[0037]

[0038] R1, R2, Z, and W are the same as above.

[0039] Thirdly, the present invention provides the application of the above-mentioned aldehyde hydrazone photocage compound in the preparation of antitumor drugs.

[0040] Fourthly, the present invention provides the application of the above-mentioned aldehyde hydrazone photocage compound in the preparation of photodynamic drugs.

[0041] Beneficial effects:

[0042] This invention provides a class of aldehyde hydrazone photocage compounds that absorb near-ultraviolet to visible light and can undergo photodegradation within a specific wavelength range, releasing the corresponding aldehyde compound. The aldehyde hydrazone compound structure described in this invention can serve as a universal strategy for the design of photorelease prodrugs with highly spatiotemporally controllable photorelease; it can be photodegraded both in vivo and in vitro; and it can also be used as an adjunct to photodynamic therapy. Attached Figure Description

[0043] Figure 1 The graph shows the changes in fluorescence intensity of the photodegradation cage in the YB31 solution and the experimental results of the photodegradation cage of the target analyte.

[0044] Figure 2 The graph shows the changes in fluorescence intensity of the photodegradation cage in solution and the experimental results of the photodegradation cages of target substances FY4 and BW3.

[0045] Figure 3 The graph shows the changes in fluorescence intensity of the photodegradation cage in solution and the experimental results of the photodegradation cages of the target compounds PC2 and TP2.

[0046] Figure 4 These are images showing enhanced fluorescence in YB7 and YB14 cells under light irradiation.

[0047] Figure 5 This is a photofluorescence enhancement image of LZ1 cells;

[0048] Figure 6 This is a photofluorescence enhancement image of CY5Q2 cells;

[0049] Figure 7 This is an imaging image of a subcutaneous xenograft tumor in a CY5Q2 mouse.

[0050] Figure 8 This is an imaging image of a subcutaneous xenograft tumor in a CYHR1 mouse.

[0051] Figure 9 This is a fluorescence intensity curve of subcutaneous xenograft tumors in CY5Q2(A) and CYHR1(B) mice. Detailed Implementation

[0052] To further illustrate the present invention and its advantages, specific embodiments are provided below to further explain the technical solution of the present invention. It should be understood that these embodiments are only helpful in understanding the present invention and should not be considered as specific limitations of the present invention. All raw materials and reagents used in the present invention are commercially available products. Unless otherwise specified, all parts mentioned in the present invention are parts by weight, and all percentages are mass percentages.

[0053] (I) Synthesis Example:

[0054] 1. The condensation reaction of stellaldehyde and hydrazine compounds was used to prepare a series of aldehyde hydrazone photocage compounds, as follows:

[0055]

[0056]

[0057] Example 1 YB7

[0058] 100 mg of stellate and 115 mg of 3,5-difluorophenylhydrazine hydrochloride were added to a 50 ml reaction flask, followed by 20 ml of a 3:1 mixture of DCM and methanol. The reaction was stopped after TLC showed no stellate. The crude product was purified by wet column chromatography, but TLC showed impurity. The crude product was then purified by slurrying with 10 ml of anhydrous ethanol. After filtration, the filter cake was purified by slurrying with 5 ml of a 15:1 mixture of DCM and methanol. After filtration, the filter cake was vacuum dried to obtain 393.2 mg of a grayish-yellow solid.

[0059] HRMS(ESI):C 35 H 31 F2N6O3,620.2347,found:([M+H] + )621.2517

[0060] 1 H NMR (600MHz, DMSO-D6) δ10.63(s,0H),9.51(d,J=1.7Hz,1H),8.13(s,1H),7.97(dd,J=8.2,6.4Hz,2H ),7.90(dd,J=8.6,1.7Hz,1H),7.62(d,J=8.5Hz,1H),7.41(ddd,J=8.5,7.0,1.4Hz,1H),7.28(t,J=7 .4Hz,1H),6.75–6.69(m,3H),6.47(ddq,J=10.4,8.1,1.7Hz,1H),4.97(s,2H),4.04(d,J=3.6Hz,1H) ,3.25(d,J=3.9Hz,1H),2.54–2.50(m,1H),2.29(d,J=4.2Hz,3H),1.42(s,3H),1.06(t,J=7.0Hz,1H).

[0061] 13C NMR(151MHz,DMSO-D6)δ172.74,172.66,164.93,164.82,163.32,163.22,148.76,14 1.34,140.03,137.15,132.94,132.88,130.54,127.75,126.96,125.10,124.96,124. 32,123.31,123.17,121.38,120.33,119.33,115.82,114.63,114.26,109.42,95.10,94.92,93.14,91.60,83.31,80.52,57.73,50.42,46.01,45.89,33.69,30.27,29.88.

[0062] Example 2 YB17

[0063] 100 mg of stellate and 142 mg of 4-bromophenylhydrazine hydrochloride were added to a 50 ml reaction flask. 20 ml of a 3:1 mixture of DCM and methanol was added and the reaction was carried out for 3 h. The reaction was stopped after TLC showed no stellate. The mixture was filtered, and the filter cake was slurried with 10 ml of ethanol and then filtered again. The filter cake was then dried under vacuum to obtain 93 mg of a grayish-yellow solid.

[0064] HRMS(ESI):C 35 H 31 BrN6O3662.1641,found([M+H] + )665.1706

[0065] 1HNMR (600MHz, DMSO-D6) δ10.35(d,J=3.6Hz,1H),9.45(d,J=3.5Hz,1H),8.55(d,J=3.4Hz,1H),8.12(d,J=3.4Hz,1H),7.97(q,J=6 .6Hz,2H),7.90(dd,J=8.7,3.5Hz,1H),7.61(dd,J=8.5,3.4Hz,1H),7.42(dd,J=7.7,3.1Hz,1H),7.40(d,J=3.2Hz,1H),7.38(d,J =3.6Hz,1H),7.28(td,J=7.4,3.4Hz,1H),7.08(d,J=3.5Hz,1H),7.06(d,J=3.4Hz,1H),6.71(q,J=3.5Hz,1H),4.97(d,J=3.4Hz,2 H),4.04(d,J=3.7Hz,1H),3.35(s,3H),3.25(d,J=8.1Hz,1H),2.52(s,2H),2.29(d,J=3.5Hz,3H),1.43(s,3H),0.86–0.62(m,1H).

[0066] 13 C NMR(151MHz,DMSO-D6)δ172.72,145.57,140.02,140.00,136.94,132.88 ,132.26,130.54,127.68,127.44,124.93,124.34,123.14,122.86,121.3 8,120.31,119.34,115.82,114.64,114.19,114.16,109.44,109.37,91.5 8,83.31,80.51,57.73,57.71,50.47,46.01,40.00,33.79,30.28,29.92.

[0067] Example 3 YB30

[0068] 100 mg of stellate and 142 mg of 2-bromophenylhydrazine hydrochloride were added to a 50 ml reaction flask, and 20 ml of a 3:1 mixture of DCM and methanol was added. The reaction was carried out for 3 h. The reaction was stopped after TLC showed no stellate. The crude product was purified by wet column chromatography, and the silica gel and product were separated by column chromatography. The liquid was evaporated to dryness to obtain 80 mg of grayish-yellow solid.

[0069] HRMS(ESI):C 35 H 31 BrN6O3,([M+H] +)662.1642, found:665.1736

[0070] 1 HNMR (600MHz, DMSO-D6) δ9.53(d,J=7.7Hz,2H),8.66(s,1H),8.57(s,1H),8.06(dd,J=8.2,5.2Hz,2H),7.96(dd,J=8.6,1.7Hz,1H),7.65–7.61(m ,2H),7.54–7.47(m,2H),7.40–7.31(m,2H),6.94–6.90(m,1H),6.76–6. 70(m,1H),5.01(s,2H),4.48(s,1H),2.45(s,3H),1.19(t,J=7.3Hz,3H).

[0071] 13 C NMR(151MHz,DMSO-D6)δ172.48,172.40,143.30,143.23,142.91,136.98,133.17,133.10,130.39,129.11,127.84,127.24,125.84,1 25.46,124.38,123.32,122.13,121.09,120.35,120.06,115.26,114.78,110.09,106.38,106.33,81.26,46.00,45.88,28.86,8.96.

[0072] Referring to the above preparation method, the following stellaria aldehyde hydrazone photocage compounds were prepared according to one embodiment of the present invention. Preferably, the following compounds are preferred among the aldehyde hydrazone photocage compounds of the present invention:

[0073]

[0074]

[0075]

[0076] 2. CY Aldehyde Hydrazone Series

[0077] Indole

[0078] CY

[0079]

[0080] CY5Q

[0081]

[0082] Example 4 CY5Q

[0083] 500 mg of trialdehyde was dissolved in 50 ml of ethanol. After complete dissolution, 2456 mg of 3-ethyl-1,1,2-trimethyl-1H-indole-3-onium iodide was added to the reaction flask. Then, 8 drops of piperidine were added (catalytic reaction), and the reaction solution turned black. The reaction was monitored by TLC (DCM:MEOH = 15:1). After four hours, the reaction was completed, and the solution was purified by column chromatography (DCM:MEOH = 60:1) to obtain 300 mg of pure blue CY5Q.

[0084] 1 H NMR(600MHz,DMSO-D6)δ10.11(s,1H),8.36–8.20(m,2H),7.74(s,2H),7.62(s,2H) ),7.50(s,2H),7.39(s,2H),7.29(s,2H),4.34(s,4H),1.76(s,12H),1.38(s,6H).

[0085] 13 C NMR (151MHz, DMSO-D6) δ177.5,142.6,141.6,129.2,126.8,123.2,120.4,112.8,50.5,40.5,40.4,40.3,40.2,40.1,40.0,39.8,39.7,27.6,12.8.

[0086] Example 5CY5Q-1:

[0087] Weigh 100 mg of CY5Q (1 equivalent) and dissolve it in 6 ml of dichloromethane. After stirring evenly, add 540 ml (3 equivalents) of phenylhydrazine to the reaction flask. React at room temperature for one hour. When the solution changes from blackish-blue to indigo-blue, monitor the end of the reaction by TLC (DCM:MEOH = 15:1). Filter the solution and purify it by TLC (DCM:MEOH = 15:1) to obtain 24 mg of green solid.

[0088] 1 H NMR (600MHz, DMSO-D6) δ10.31(s,1H),8.39(s,1H),8.14(d,J=14.4Hz,2H),7.71–7.66(m,2H),7.31(td,J=7.3,1.1Hz,2H),7.27( dd,J=8.4,7.2Hz,2H),7.10–7.03(m,4H),6.79(tt,J=7.3,1.2Hz,1H),4.24(q,J=7.2Hz,4H),1.77(s,11H),1.36(t,J=7.2Hz,6H).

[0089] 13 C NMR (151MHz, DMSO-D6) δ174.4,145.7,142.1,141.9,129.7,129.0,125.7,123.0,119.3,11 2.1,111.8,101.0,49.8,40.4,40.3,40.2,40.0,39.9,39.8,39.6,39.5,29.5,27.4,12.7.

[0090] Example 6CY5Q-2:

[0091] Weigh 100 mg of CY5Q (1 equivalent) and dissolve it in 6 ml of dichloromethane. After stirring evenly, add 103.9 mg (3 equivalents) of 4-carboxylic acid phenylhydrazine to the reaction flask. React at room temperature for one hour. When the solution changes from blackish-blue to indigo-blue, the reaction is considered complete by TLC (DCM:MEOH = 15:1). Filter the solution and purify it by TLC (DCM:MEOH = 8:1) to obtain 43.9 mg of green solid.

[0092] 1 H NMR (400MHz, DMSO-D6) δ12.37(s,1H),10.72(s,1H),8.43(s,1H),8.17(d,J=14.5Hz,2H),7.86(d,J=8.5Hz,2H),7.69(d,J=7.4Hz,2H),7.53–7 .41(m,4H),7.36–7.28(m,2H),7.16–7.02(m,4H),7.03(s,1H),6.72–6 .59(m,1H),4.26(q,J=7.2Hz,4H),1.77(s,12H),1.37(t,J=7.2Hz,6H).

[0093] 13 C NMR (101MHz, DMSO-D6) δ174.7,167.7,160.5,149.0,145.4,142.1,141.8,137.6,131.7,129.0,128.0,125. 8,123.1,122.3,120.7,118.6,111.9,111.2,105.6,101.0,74.0,49.9,44.1,36.3,30.2,27.4,12.7,10.9.

[0094] Example 7CY5Q-3:

[0095] Weigh 100 mg of CY5Q (1 equivalent) and dissolve it in 6 ml of dichloromethane. After stirring evenly, add 64.74 mg (3 equivalents) of nitrophenylhydrazine to the reaction flask. React at room temperature for one hour. When the solution changes from blackish-blue to indigo-blue, the reaction is monitored by TLC (DCM:MEOH = 15:1) to indicate the end of the reaction. Filter the solution to obtain 30 mg of green filter cake.

[0096] 1 H NMR (400MHz, DMSO-D6) δ11.46(s,1H),8.64(s,1H),8.24–8.14(m,4H),7.70(d,J=7.4Hz,2H),7.48(dt,J=15.1,7.9Hz,4H),7.33(t ,J=7.3Hz,2H),7.15(d,J=8.8Hz,2H),7.05(d,J=14.5Hz,2H),4.27(q,J=7.2Hz,4H),1.77(d,J=1.5Hz,12H),1.37(t,J=7.1Hz,6H).

[0097] 13 C NMR (101MHz, DMSO-D6) δ175.0,152.9,150.7,142.2,141.8,140.3,138.6,129.1,126.8,126.0,12 3.1,120.8,112.0,111.2,101.2,50.0,40.7,40.5,40.3,40.0,39.8,39.7,39.6,39.4,27.4,12.8.

[0098] Benzindole

[0099] CY

[0100]

[0101] Example 8: Synthesis of Cy5A

[0102] 558 mg (5.58 mmol) of trialdehyde and 3-ethyl-1,1,2-trimethyl-1H-benzo[e]indole-3-iodide (2.04 g, 11.18 mmol) were placed in a 250 mL round-bottom flask with methanol (40 mL). The reaction mixture was stirred at room temperature (30 °C) for 10 hours. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 100:1) to give a blue-black solid Cy5A (518 mg, 0.96 mmol), in a yield of 17.3%.

[0103] 1H NMR (400MHz, CDCl3) δ7.41(h,J=3.5Hz,1H),6.67(d,J=8.6Hz,2H),6.38(d,J=2.6Hz,2H),6.27(dd ,J=8.7,2.6Hz,2H),5.35(s,1H),4.59–4.54(m,2H),3.32(q,J=7.1Hz,8H),1.15(t,J=7.0Hz,12H); 13 C NMR (101MHz, CDCl3) δ151.68,147.82,144.76,138.61,132.40,131.40,129.9 3,128.00,127.00(d,J=20.7Hz),107.50,98.87,63.01,44.39,40.00,12.61.

[0104] Example 9IIa: Synthesis of CYHR1

[0105] Cy5A (100 mg, 0.18 mmol) and 4-hydrazinoic acid (42.3 mg, 0.28 mmol) were reacted in a 50 mL round-bottom flask containing 15 mL of methanol at room temperature (30 °C) for 2 hours. After reaction, the mixture was filtered and dried to give a dark green solid CYHR1 (80 mg, 0.12 mmol), with a yield of 66.7%.

[0106] 1 H NMR(400MHz, CDCl3)δ7.28(d,J=7.1Hz,1H),7.18(d,J=7.3Hz,1H),7.14–7.04(d, J=7.1Hz,1H),7.03–6.87(d,J=7.3Hz,2H),6.84(d,J=8.7Hz,1H),6.75(dd,J=20. 9,8.4Hz,1H),6.47(d,J=8.3Hz,1H),5.64(s,1H),5.22(d,J=11.5Hz,2H),3.59(d ,J=6.6Hz,4H),3.54–3.43(m,4H),3.43(q,J=7.1Hz,8H),1.15(q,J=4.5Hz,12H), 13C NMR (101MHz, CDCl3) δ172.94,152.69,151.01,147.10,140.67,134.09,130.63,130.52,129.33,129.02,128.82,128. 77,128.57,128.27,123.20,123.03,113.34,108.17,100.84,66.00,56.15,53.71,44.50,40.34,39.71,38.06,12.35.

[0107] Example 10IIb: Synthesis of CYHR2

[0108] Cy5A (50 mg, 0.09 mmol) and 4-nitrophenylhydrazine (105.4 mg, 0.56 mmol) were reacted in a 50 mL round-bottom flask containing 7 mL of methanol at room temperature (30 °C) for 0.5 h. The reaction mixture was filtered and dried to give a dark green solid CYHR2 (41.6 mg, 0.06 mmol), in a yield of 66.7%.

[0109] 1 H NMR (400MHz, CDCl3) δ8.19(d,J=7.0Hz,2H),7.50(d,J=7.5,2.0Hz,2H),7.46–7.40(d,J=2.1H z,2H),7.36–7.24(m,1H),7.13(d,J=8.5,1.6Hz,2H),7.06–6.99(m,1H),6.90(d,J=1.8Hz,1H) ,6.85(d,J=8.1,2.2Hz,1H),6.66–6.60(d,J,2H),5.33(S,2H),3.83(S,1H),3.69(t,J=3.4Hz ,4H),3.60(t,J=3.4Hz,4H),3.45(q,J=7.0Hz,8H),3.16–3.03(m,2H),1.11(t,J=7.1Hz,12H), 13C NMR (101MHz, CDCl3) δ172.91,157.60,155.40,154.64,153.54,147.10,139.64,134.64,134.46,132.92,130.54,130.52,129.75,129.64,128 .47,128.27,128.00,115.71,115.70,113.38,113.34,113.31,95.85, 95.03,66.81,56.15,53.71,45.26,44.95,40.34,39.71,12.35,11.78.

[0110] Coumarin Series:

[0111]

[0112] Example 11

[0113] 50 mg of 7-(diethylamine)coumarin-3-carboxaldehyde and 65 mg of phenylhydrazine were dissolved in a 50 ml round-bottom flask equipped with a stirrer and 5 ml of dichloromethane. The reaction temperature was controlled at 25 °C and the rotation speed at 500 rpm. -1 The reaction was stirred for 3 hours, and the reaction was monitored by TLC thin-layer plate during the process. The mixture was filtered through an ethanol heater and the filter cake was washed to obtain the target product LG05 3.1 mg. ¹H NMR (500 MHz, DMSO) δ 10.50 (s, 1H), 8.25 (s, 1H), 7.90 (s, 1H), 7.59 (dd, J = 8.9, 1.6 Hz, 1H), 7.23 (td, J = 8.1, 1.7 Hz, 2H), 7.11–7.05 (m, 2H), 6.79–6.71 (m, 2H), 6.57 (d, J = 2.4 Hz, 1H), 3.46 (q, J = 7.1 Hz, 4H), 1.14 (t, J = 7.0 Hz, 6H). ¹³C NMR (126MHz, DMSO) δ 161.39, 156.03, 150.76, 145.53, 135.45, 131.18, 130.34, 129.57, 119.30, 115.09, 112.45, 112.42, 109.96, 108.94, 96.96, 44.60, 12.86. ESI-MS was LG0, theoretical value was 335.1634, result was ([M+H] + )336.1704.

[0114] Example 12

[0115] 50 mg of 7-(diethylamino)coumarin-3-carboxaldehyde and 114 mg of 2-nitrophenylhydrazine were dissolved in a 50 ml round-bottom flask equipped with a stirrer and 5 ml of dichloromethane, and 84 μL of triethylamine was added as a catalyst. The reaction temperature was controlled at 25 °C and the rotation speed was 500 rpm. -1 The reaction was stirred for 3 hours, and the reaction was monitored by TLC during the process. The filter cake was filtered and washed with an ethanol heater to obtain the target product LG1 65.4 mg. 1 H NMR (500MHz, DMSO) δ11.32 (s, 1H), 8.49 (s, 1H), 8.45 (s, 1H), 8.11 (dd, J=8.6, 1.6Hz, 1H), 8.04 (d, J=8.7Hz, 1H), 7.64 (dd, J=26.1, 8.3Hz, 2H), 6.92 (t, J=7.7Hz, 1H), 6.78 (dd, J=9.0, 2.4Hz, 1H), 6.59 (d, J=2.4Hz, 1H), 3.48 (q, J=7.0Hz, 4H), 1.15 (t, J=7.0Hz, 6H). 13 C NMR (126MHz, DMSO) δ 161.02, 156.79, 151.59, 141.62, 140.09, 138.75, 136.63, 131.16, 131.01, 126.13, 118.65, 116.75, 113.63, 110.18, 108.74, 96.92, 44.70, 12.86. ESI-MS was LG1, theoretical value was 380.1485, result was ([M+H] + )381.1564.

[0116] Example 13

[0117] 50 mg of (diethylamine)coumarin-3-carboxaldehyde and 114 mg of 4-nitrobenzene were dissolved in a 50 ml round-bottom flask equipped with a stirrer and 5 ml of dichloromethane, and 84 μL of triethylamine was added as a catalyst. The reaction temperature was controlled at 25 °C and the rotation speed at 500 rpm. -1 The reaction was stirred for 3 hours, and the reaction was monitored by TLC during the process. The filter cake was filtered and washed with an ethanol heater to obtain the target product LG357.9 mg. 1H NMR (500MHz, DMSO) δ8.39 (s, 1H), 8.14 (d, J = 8.9Hz, 1H), 8.09 (s, 1H), 7.61 (d, J = 8.8 Hz, 1H), 7.18 (d, J=8.4Hz, 1H), 6.78 (dd, J=9.0, 2.4Hz, 1H), 6.59 (d, J=2.4Hz, 1H), 3 0.48 (q, J = 7.0 Hz, 3H), 3.31 (s, 6H), 1.15 (t, J = 7.0 Hz, 5H). 13C NMR (126 MHz, DMSO) δ 126.65, 110.21, 108.78, 46.15, 44.69, 12.86. ESI-MS was LG3, theoretical value was 380.1485, result was ([M+H] + )381.1577.

[0118] TPA derivatives

[0119]

[0120] Example 14

[0121] 100 mg of 4-diphenylaminobenzaldehyde and 208 mg of 2-nitrophenylhydrazine were dissolved in a 50 mL round-bottom flask equipped with a stirrer and 10 mL of dichloromethane, and 152 mL of triethylamine was added as a catalyst. The reaction was controlled at 25 °C and stirred at 500 rpm for 3 hours, with the reaction monitored by TLC. The mixture was filtered through an ethanol heater and the filter cake was washed to obtain the target product SA0. 1 H NMR (500MHz, DMSO) δ11.16 (s, 1H), 8.43 (s, 1H), 8.11 (dd, J=8.6, 1.5Hz, 1H), 7.94 (dd, J=8.7, 1.3Hz, 1H), 7. 68–7.61 (m, 3H), 7.40–7.32 (m, 4H), 7.16–7.06 (m, 6H), 7.01–6.95 (m, 2H), 6.89 (ddd, J=8.4, 6.9, 1.3Hz, 1H). 13 CNMR (126MHz, DMSO) δ 149.08, 147.08, 145.58, 141.93, 136.83, 130.89, 130.20, 128.57, 128.44, 126.16, 125.26, 124.36, 122.30, 118.36, 116.41. ESI-MS is SA0, theoretical value is 408.1586, result is ([M+H] + )409.1654.

[0122] Example 15

[0123] 100 mg of 4-diphenylaminobenzaldehyde and 208 mg of 2-bromo-5-hydrazidepyridine were dissolved in a 50 mL round-bottom flask equipped with a stirrer and 10 mL of dichloromethane. The reaction temperature was controlled at 25 °C and the rotation speed at 500 rpm. -1 The reaction was stirred for 3 hours, and the reaction was monitored by TLC during the process. The filter cake was then filtered and washed with an ethanol heater to obtain the target product SA4.

[0124] 1 H NMR (600MHz, DMSO-D6) δ10.94 (s, 1H), 8.12 (dd, J=14.4, 2.4Hz, 1H), 7.98 (d, J=2.5Hz, 2H), 7.93 (s, 1H), 7.72 (ddd, J=17.4, 8.9, 2.6Hz, 1H), 7.65 ( s, 2H), 7.58–7.49 (m, 3H), 7.32–7.25 (m, 2H), 7.12 (d, J=8.9Hz, 1H), 7.07 –6.95(m, 4H), 6.91(d, J=8.5Hz, 1H), 6.65(d, J=9.0Hz, 2H), 4.16(s, 1H). 13 C NMR (151MHz, DMSO-D6) δ 161.26, 148.01, 139.61, 130.20, 127.85, 125.02, 124.13, 122.87, 108.65, 106.01. ESI-MS was SA4, theoretical value 442.0793, result ([M+H] + )443.0865.

[0125] Example 16

[0126] 100 mg of 4-diphenylaminobenzaldehyde and 159 mg of 2-chloro-6-hydrazidepyridine were dissolved in a 50 mL round-bottom flask equipped with a stirrer and 10 mL of dichloromethane. The reaction temperature was controlled at 25 °C, the stirring speed at 500 r·s⁻¹, and the reaction was stirred for 3 hours, with the reaction monitored by TLC thin-layer plate during the process. ¹H NMR (500 MHz, DMSO) δ 11.15 (s, 1H), 7.96 (s, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.56 (d, J = 8.2 Hz, 2H), 7.34 (t, J = 7.6 Hz, 4H), 7.10 (dt, J = 22.9, 8.8 Hz, 7H), 6.96 (d, J = 8.2 Hz, 2H), 6.78 (d, J = 7.5 Hz, 1H). ¹³C NMR (126MHz, DMSO) δ 157.70, 148.64, 148.41, 147.22, 141.45, 140.77, 130.15, 129.13, 127.90, 125.04, 124.13, 122.66, 113.84, 105.16. ESI-MS was SA5, theoretical value was 398.1298, result was ([M+H] + )399.1385.

[0127] TP1

[0128]

[0129] Example 17

[0130] 4-(1,2,2-triphenylvinyl)benzaldehyde (2 mmol) was reacted with (4-bromophenyl)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 60.4%. 1 HNMR (400MHz, DMSO-d6) δ10.40(s,1H),7.73(s,1H),7.38–7.33(m,2H),7.31–7.26(m,2H),7.12–6.91(m,19H). 13 C NMR (101MHz, DMSO-D6) δ145.0,143.7,143.7,143.6,143.5,141.3,140.8,137.5,134.2,1 32.2,131.6,131.3,131.2,128.5,128.4,128.3,127.2,125.7,114.4,110.1.HRMS(ESI):

[0131] C33H25BrN2,([M+H]+)530.1181,found:529.1259.

[0132] Example 18TP2

[0133] 4-(1,2,2-triphenylvinyl)benzaldehyde (2 mmol) was reacted with (4-nitrobenzene)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 55.9%. 1 HNMR (600MHz, DMSO-d6) δ11.24(s,1H),8.07(d,J=8.9Hz,2H),7.91(s,1H),7.44(d,J=8.0Hz,2H),7.10(d,J=9.8Hz,10H),6.95(q,J=8.3,7.7Hz,9H). 13 C NMR (151MHz, DMSO-D6) δ151.1,144.7,143.7,143.6,143.4,142.0,141.7,140.7,138.9,133.4,131.7,131.3,131.2,131. 2,128.5,128.4,128.4,127.3,127.2,126.7,126.5,111.8.HRMS(ESI):C33H25N3O2,([M+H]+)496.1980,found:496.2018.

[0134]

[0135] Example 19FY1

[0136] Anthracene-9-carboxaldehyde (2 mmol) was reacted with (3-bromophenyl)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 60.5%. 1HNMR(600MHz,DMSO-d6)δ10.89(s,1H),9.07(s,1H),8.65(dd,J=8.9,1.2Hz,2H),8.58(s,1H),8.09(dd,J=8.5,1.3Hz,2H), 7.58(ddd,J=8.9,6.5,1.4Hz,2H),7.52(ddd,J=7.8,6.5,1.1Hz,2H),7.25(t,J=2.0Hz,1H),7.18(t,J=8.0Hz,1H),7.05(ddd ,J=8.2,2.2,1.0Hz,1H),6.91(ddd,J=7.8,2.0,1.0Hz,1H).13CNMR(151MHz,DMSO-D6)δ147.4,137.2,131.8,131.6,129.7,1 29.6,129.5,128.6,127.3,127.2126.9,126.2,126.0,125.9,125.6,125.4,125.3,123.1,121.8,114.7,111.6.HRMS(ESI):

[0137] C21H15BrN2,([M+H]+)376.0398,found:375.0495.

[0138] Example 20FY2

[0139] Anthracene-9-carboxaldehyde (2 mmol) was reacted with (4-fluorophenyl)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 73.4%. 1 HNMR(600MHz,DMSO-d6)δ10.78(s,1H),9.08(s,1H),8.70(d,J=8.9Hz,2H),8.54(s,1H),8 .07(d,J=8.4Hz,2H),7.57(t,J=7.6Hz,2H),7.51(t,J=7.4Hz,2H),7.09(d,J=6.8Hz,4H). 13C NMR (151MHz, DMSO-D6) δ157.3,155.8,142.6,135.8,131.7,129.6,129.44,128.2,127.2,127.1,126.1,125 .9,125.7,125.6,116.4,116.3,113.5,113.4.HRMS(ESI):C21H15FN2,([M+H]+)315.1253,found:315.1310.

[0140] Example 21FY3

[0141] Anthracene-9-carboxaldehyde (2 mmol) was reacted with phenylhydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 74.8%. 1 HNMR (600MHz, DMSO-d6) δ10.65(s,1H),9.06(s,1H),8.72(d,J=8.9Hz,2H),8.54(s,1H),8.07(d,J=8.4Hz,2H),7.58(d,J =6.5Hz,1H),7.58-7.55(m,1H),7.51(dd,J=8.3,6.5Hz,2H),7.30-7.22(m,2H),7.11(d,J=8.0Hz,2H),6.80-6.74(m,1H). 13 C NMR(151MHz,DMSO-D6)δ145.8,135.7,131.7,129.9,129.6,129.5,128.2,127.2,127.1, 125.9,125.6,119.6,112.5.HRMS(ESI):C21H16N2,([M+H]+)297.1347,found:297.1384.

[0142] Example 22FY4

[0143] Anthracene-9-carboxaldehyde (2 mmol) was reacted with 4-hydrazinobenzoic acid (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 66.1%. 1HNMR(600MHz,DMSO-d6)δ11.10(s,1H),9.14(s,1H),8.69(d,J=8.9Hz,2H),8.58(s,1H),8.09(d,J=8.4Hz,2H) ,7.86(d,J=8.3Hz,2H),7.60(dd,J=9.0,6.3Hz,2H),7.52(t,J=7.4Hz,2H),7.14(d,J=8.4Hz,2H),5.71(s,1H). 13 C NMR (151MHz, DMSO-D6) δ167.8,149.3,138.3,131.9,131.6,131.4,129.8,129.5,128.8 ,127.4,126.7,126.0,125.4,121.2,111.78,110.4.HRMS(ESI):C22H16N2O2,([M+H]+)

[0144] 341.1245, found: 341.1273.

[0145]

[0146] Example 23BW1

[0147] 1-Pyrenecarbaldehyde (2 mmol) was reacted with (3-bromophenyl)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 76.2%. 1 HNMR (400MHz, DMSO-D6) δ10.86(s,1H),8.91(s,1H),8.68(d,J=9.4Hz,1H),8.54(d,J=8.1Hz,1H),8.34–8.21(m,4H),8.21–8.09(m,2H),8.09–7. 98(m,1H),7.36(t,J=2.0Hz,1H),7.26–7.14(m,1H),7.12(ddd,J=8.2,2.1,1.1Hz,1H),6.92(ddd,J=7.7,2.0,1.0Hz,1H),2.46(p,J=1.8Hz,2H). 13C NMR (101MHz, DMSO-D6) δ147.3,137.0,131.7,131.6,131.2,130.9,128.9,128.7,128.1,12 8.0,127.8,127.0,126.2,125.8,125.8,124.9,124.6,123.1,123.0,121.9,114.8,111.8.

[0148] Example 24BW2

[0149] 1-Pyrenecarbaldehyde (2 mmol) was reacted with (3-phenylethyl)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 65.8%. 1 HNMR (600MHz, DMSO-d6) δ10.55(s,1H),8.84(s,1H),8.72(d,J=9.4Hz,1H),8.52(d,J=8.1Hz,1H),8.25 (d,J=8.1Hz,4H),8.12(s,2H),8.04(d,J=7.7Hz,1H),7.10(s,4H),2.46(s,2H),1.13(t,J=7.7Hz,3H). 13 CNMR(151MHz,DMSO-D6)δ143.7,134.9,134.9,131.7,131.0,130.8,129.6,129.1,128.5,128 .0,127.7,127.4,126.9,126.0,125.8,125.6,125.0,124.7,124.4,123.2,112.8,28.0,16.5.

[0150] Example 25BW3

[0151] 1-Pyrenecarboxaldehyde (2 mmol) was reacted with 4-hydrazinobenzoic acid (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 71.2%. 1HNMR (400MHz, DMSO-D6) δ11.08(s,1H),8.97(s,1H),8.70(d,J=9.3Hz,1H),8.57(d,J=8.3Hz,1H),8. 28(d,J=7.9Hz,5H),8.15(s,2H),8.05(t,J=7.7Hz,1H),7.87(d,J=8.4Hz,2H),7.23(d,J=8.2Hz,2H). 13 CNMR(101MHz,DMSO-D6)δ167.9,149.2,138.1,131.9,131.6,131.4,130.8,128.8,128.7,1 28.2,128.0,127.0,126.3,125.9,125.8,124.9,124.7,124.6,123.0,121.2,112.0,110.3.

[0152]

[0153] Example 26CM1

[0154] (E)-3-(4-(dimethylamino)phenyl)propenal (2 mmol) was reacted with 4-hydrazinonitrile (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 67.5%. 1 H NMR (400MHz, DMSO-D6) δ10.71 (s, 1H), 7.74 (dd, J = 6.8, 1.4Hz, 1H), 7.53 (d, J = 8. 5Hz,2H),7.40–7.31(m,2H),7.05–6.95(m,2H),6.79–6.60(m,4H),2.89(s,6H). 13 C NMR (101MHz, DMSO-D6) δ150.8,149.0,144.5,137.0,134.1,133.9,128.5,124.7,121.4,120.8,113.3,112.7,112.3,99.1,40.4.

[0155] Example 27CM2

[0156] (E)-3-(4-(dimethylamino)phenyl)propenal (2 mmol) was reacted with (3-nitrobenzene)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 58.4%. 1 H NMR (400MHz, DMSO-D6) δ10.55(s,1H),7.76-7.67(m,2H),7.47(dd,J=8.0,2.4Hz,1H),7.44-7.32(m ,3H),7.25(d,J=7.1Hz,1H),6.82-6.71(m,2H),6.66(dd,J=12.6,9.0Hz,2H),2.89(d,J=3.4Hz,5H). 13 C NMR (101MHz, DMSO-D6) δ150.8,149.4,147.0,143.5,136.5,130.8,128.4,124.8,121.6,118.5,112.8,112.7,105.7,40.4.

[0157] Example 28CM3

[0158] (E)-3-(4-(dimethylamino)phenyl)propenal (2 mmol) was reacted with 3-chloro-6-hydrazidopyridazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 61.8%. 1 H NMR (400MHz, DMSO-D6) δ11.40(s,1H),7.88(d,J=8.8Hz,1H),7.57(d,J=9.2Hz,1H),7.37(t,J=8.0H z,3H),6.77(d,J=15.8Hz,1H),6.68(dd,J=18.8,8.4Hz,3H),3.33(s,3H),3.26(s,2H),2.90(s,5H). 13 C NMR (101MHz, DMSO-D6) δ159.2,137.7,130.1,128.6,121.1,116.6,112.6,40.4.

[0159]

[0160] Example 29PC1

[0161] 3-Perenal (2 mmol) was reacted with (3-bromophenyl)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 74.8%. 1 HNMR(600MHz,DMSO-d6)δ10.80(s,NH),8.52(s,Ar-H),8.49(d,J=8.5Hz,Ar-H),8.41-8.36(m,A r-H),8.36-8.32(m,2Ar-H),8.32-8.29(m,Ar-H),7.91(d,J=8.0Hz,Ar-H),7.76(d,J=8.0Hz,2Ar -H),7.62(dd,J=8.5,7.5Hz,Ar-H),7.51(td,J=7.8,1.7Hz,2Ar-H),7.29(t,J=2.0Hz,Ar-H),7. 18(t,J=7.9Hz,Ar-H), 7.07(ddd,J=8.3,2.1,1.0Hz,Ar-H), 6.91(ddd,J=7.7,2.0,0.9Hz,Ar-H). 13 C NMR(151MHz,DMSO-d6)δ147.2,137.5,134.8,131.8,131.7,131.5,131.1,130.9,130.8,129.0,128.7,1 28.6,128.1,128.0,127.6,126.6,124.3,123.1,121.8,121.6,121.5,121.5,121.2,114.8,111.7,46.0.

[0162] Example 30PC2

[0163] 3-Perenal (2 mmol) was reacted with (3-nitrobenzene)hydrazine (2 mmol) in DCM solution (6 mmol) and stirred in the dark for 36 hours to obtain a crude product. The crude product was washed successively with DCM and water, and purified by silica gel column chromatography. The eluent was eluted with (DCM / MeOH = 50:1), and the eluent was rotary evaporated to obtain the target product in a yield of 79.6%. 1HNMR (400MHz, DMSO-d6) δ11.10(s,1H),8.56(s,1H),8.48(d,J=8.5Hz,1H),8.40(d,J=7.5Hz,1H),8.37–8.32(m,3H),7.93(d,J=8.1Hz,1H), 7.77(d,J=8.1Hz,2H),7.64(t,J=8.0Hz,1H),7.55–7.49(m,2H),6.73–6.70(m,1H),6.63(dt,J=7.7,2.1Hz,2H),6.49(tt,J=9.4,2.4Hz,1H). 13 C NMR (101MHz, DMSO-D6) δ149.4,148.3,138.5,134.7,131.7,131.5,131.4,131.0,130.8,130.5,128.9 ,128.8,128.6,128.1,127.6,127.0,124.2,121.7,121.6,121.2,98.0,97.7,96.2,95.6,95.3,94.0.

[0164] The synthetic routes for the Rhodamine series are as follows:

[0165]

[0166]

[0167] Example 31LZ1

[0168] Synthesis of Rh6G-OH. 6 g (2.23 g) of rhodamine was added to 50 mL of dry THF and stirred in an ice bath at 0 °C for 10 min. Then, 1.9 g of lithium aluminum hydride was slowly added to the mixture, and the mixture was stirred at 0 °C for another 30 min. The reaction was monitored using thin-layer chromatography (HP-TLC silica gel 60 g 254 plate, DCM:EtOH, v / v = 10:1). After the reaction was complete, the mixture was poured into water (200 mL), and the red solid was collected by filtration. The crude product was subjected to column chromatography (silica gel, DCM:EtOH, v / v = 15:1) to obtain Rh6G-OH. The yield of the red powder was 59% (1.19 g).

[0169] Synthesis of Rh6G-CHO. Rh6G-OH (40.4 mg) and pyridine chlorochromate (40.6 mg) were dissolved in 25.0 mL of DCM. The mixture was stirred under N2 at room temperature for 2 hours, and filtered to obtain a red solid. The crude product was purified by solid silica gel column chromatography (DCM:EtOH, v / v = 15:1) to obtain a red solid Rh6G-CHO (11 mg, yield 27%).

[0170] Synthesis of LZ1. Compound Rh6G-CHO (0.13 mmol, 52.0 mg) was dissolved in 5 mL of DCM. Phenylated hydrazine (0.39 mmol) was added sequentially. After the reaction was complete, the mixture was cooled and filtered to obtain solid LZ1 (50.0 mg, yield 78%).

[0171] 1 H NMR (400MHz, DMSO-D6) δ10.29(s,1H),7.93(d,J=8.0Hz,1H),7.79(t,J=6.1Hz, 2H),7.63(t,J=7.7Hz,1H),7.54(s,1H),7.49(t,J=7.7Hz,1H),7.25(d,J=7.6Hz ,1H),6.98(t,J=7.7Hz,2H),6.90(s,2H),6.81(s,2H),6.60(t,J=7.5Hz,1H),6 .43(d,J=8.0Hz,2H),3.44(p,J=6.9Hz,4H),2.04(s,6H),1.18(d,J=7.4Hz,6H). 13 C NMR (101MHz, DMSO-D6) δ157.50,157.26,156.38,145.09,134.60,134.07,130.53,130.18,129.75,129.4 1,129.22,128.28,127.44,126.07,119.57,113.65,112.06,94.05,38.44,18.15,14.13.HRMS(ESI),m / z 489.2649,found489.2658.

[0172] Example 32

[0173] Synthesis of LZ2. LZ2 was prepared by replacing phenylhydrazine with 0.39 mmol of 2-nitrobenzylhydrazine, using a method similar to that of LZ1. The product was a red solid (20.9 mg, 30% yield).

[0174] Compound LZ2 1 H NMR (400MHz, DMSO-D6) δ10.91 (s, 1H), 8.20 (s, 1H), 8.09 (d, J = 7.9Hz,

[0175] 1H),7.95(dd,J=8.5,1.5Hz,1H),7.81(q,J=8.3,6.6Hz,2H),7.66(dtd,J=36.6,7.5,1.3Hz,2H),7.51–7.26(m,2H),7.07(dd,J=8.7, 1.3Hz,1H),6.92(s,2H),6.87–6.82(m,1H),6.80(s,2H),3.46(p,J=7.5,6.9Hz,3H),3.34(s,1H),2.05(s,6H),1.20(t,J=7.1Hz,6H). 13 C NMR(101MHz,DMSO-D6)δ157.51,156.45,156.17,142.55,141.20,136.28,133.64,131.49,131.27,130.71, 130.03,129.18,128.29,126.16,123.86,119.19,115.93,113.83,94.11,38.47,18.13,14.14.HRMS(ESI):C 32 H 32 N5O3 + ,m / z534.2500,found534.2505.

[0176] Example 33

[0177] Synthesis of LZ3. Compound Rh6G-CHO (0.13 mmol, 52.0 mg) was dissolved in 5 mL of DCM. Compound phenylhydrazine (0.39 mmol) was added sequentially. After the reaction was complete, the mixture was cooled and filtered to obtain solid LZ1 (50.0 mg, yield 78%). HRMS (ESI), m / z 553.2547, found 553.2576.

[0178] (II) Photolysis Cage Experiment

[0179] The compounds prepared in Examples 1-33 above were subjected to photodegradation experiments. The results showed that the aldehyde hydrazone photocage compounds of the present invention absorb near-ultraviolet to visible light and can undergo photodegradation within a specific wavelength range, releasing the corresponding aldehyde compounds, as exemplified below.

[0180] Example 34

[0181] 4.0 mL of a DMSO solution containing the photocage compound YB31 (100 μM) synthesized in this invention was added to a flat-bottomed quartz tube (bottom area 2.270 cm²), and the tube was irradiated with 365 nm light under water-cooled conditions at room temperature. A current stabilizer was used as the light source. The incident light intensity was measured using a power meter (WATTCAS WP-TEC-1020SL) to maintain 120.3 mW / cm². 2 ——121.6mW / cm 2 After photodegradation, samples were taken for fluorescence intensity and LC-MS analysis. Figure 1 As can be seen, the characteristic absorption of YB31 decreases with increasing irradiation time, while the wavelength and intensity of emitted light increase with increasing irradiation time. The LC-MS curve shows that YB31 decreases with increasing illumination time, while STSQ increases simultaneously, proving the photocage uncoiling process.

[0182] Example 35

[0183] Using flat-bottomed quartz tubes, 4.0 mL of a DMSO solution containing the photocage compounds FY4 and BW3 (100 μM) synthesized in this invention was added to each tube, and the reaction was carried out at 405 nm (incident light intensity 122.0 mW / cm²) under water-cooled conditions at room temperature. 2 —123.5mW / cm 2 Irradiation with light from PC2 and TP2 (100μM). Separately, PC2 and TP2 were used with light at 510nm (incident light intensity 121.7mW / cm²). 2 —123.2mW / cm 2 Irradiation by a light source. After photodegradation, samples were taken for fluorescence intensity and LC-MS analysis. Figures 2-3 The emission wavelengths of FY4, BW3, PC2, and TP2 changed significantly, and the emission intensity increased with the increase of irradiation time. The LC-MS curves showed that the concentrations of FY4, BW3, PC2, and TP2 all decreased with irradiation, while the corresponding aldehydes FYA, BWA, PCA, and TPEA increased simultaneously, further proving the photocage decongestion process.

[0184] Cell imaging experiments

[0185] Main reagents

[0186]

[0187] All cancer cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.

[0188] 1. Cell Culture

[0189] Cells were cultured in DMEM medium containing 10% FBS and 2% penicillin / streptomycin antibiotics. Cell growth was maintained in a 37°C, 5% CO2 saturated humidity incubator. After cell resuscitation, passage, seeding, and trypsin digestion, the cell density was adjusted to 5 × 10⁶ cells / year. 4 Cells / mL were seeded into 12-well plates with laser confocal slides and incubated overnight at 37°C in a 5% CO2 saturated humidity incubator for cell imaging.

[0190] 2. Cell imaging

[0191] The cells to be used were washed twice with PBS, treated with 0.1 mM oleic acid for 24 h, washed twice with PBS, stained with photocage compound (1 μM) for 30 min, washed twice with PBS, and fixed with 4% paraformaldehyde for 10 min. After washing twice with PBS, anti-fluorescence quenching mounting medium was dropped onto a glass slide, the cell slide was gently inverted on it, and placed in a humidified chamber. The cells were irradiated at 405 nm for different times using a live cell workstation and observed using a confocal microscope (Leica, STELLARIS 5, Germany).

[0192] Cell imaging experiments were performed on the compounds prepared in Examples 1-33 above. The cell imaging showed a significant increase in fluorescence intensity and range, indicating that aldehydes were released from the cells after light exposure and gradually increased and diffused, as shown in the following examples.

[0193] Example 36

[0194] After culturing MCF-7 cells for 24 hours, the cells were irradiated with DMSO solution containing 10 μM MYB7 and YB14 according to the cell imaging procedure described above. Parameters were acquired and images were obtained using a microscope after 365 nm ultraviolet light irradiation. Figure 4 The image shows a significant increase in fluorescence intensity in the blue-green channels, indicating that stellate aldehyde was released from the cells after illumination and its levels gradually increased.

[0195] Example 37

[0196] After culturing MCF-7 cells for 24 hours, the cells were irradiated with 10 μM LZ1 DMSO solution according to the cell imaging procedure described above, and parameters and images were acquired using a microscope under 530 nm visible light. Figure 5 The images show a significant increase in the intensity and range of cell fluorescence, indicating that aldehydes are released from the cells after illumination and gradually increase and diffuse.

[0197] Example 38

[0198] After culturing MCF-7 cells for 24 hours, they were irradiated with 10 μM CY5Q2 DMSO solution according to the cell imaging procedure described above, and parameters and images were acquired using a microscope. Figure 6 The fluorescence intensity of the cells and the fluorescence fold curves in the image show a significant increase, indicating that aldehydes are released from the cells after light exposure and gradually increase in amount.

[0199] In vitro cancer cell inhibition experiment

[0200] The stellaria aldehyde derivatives prepared in the above examples were subjected to in vitro cancer cell inhibition experiments to evaluate the inhibitory effect of the drug on cancer cell growth before and after photodegradation. The results showed that the photocage compound of the present invention exhibited significantly improved inhibitory performance against the cytotoxicity of the original drug, as illustrated below. Experimental methods:

[0201] Cultured cancer cells were seeded into 96-well plates, divided into two groups: one group received treatment with the drug before photodegradation (control group), and the other group received treatment with the drug after photodegradation (experimental group). After drug addition, a photoperiod was initiated, followed by continued culture in the dark for 24 hours. Cell proliferation was assessed using a CCK8 assay. Cells were collected at set time points (10 minutes, 20 minutes, and 30 minutes) during treatment and culture. The MTT assay was used to evaluate cell proliferation and viability. Experimental data were collected, statistically analyzed, and differences between the control and experimental groups were compared. The data from the control and experimental groups were compared to evaluate the inhibitory effect of the drug on cancer cell growth before and after photodegradation.

[0202] Example 39

[0203] The photocage compounds YB1, YB7, YB10, YB17, YB25, and YB30 were prepared into 10 μM DMSO solutions and cultured in MCF-7 and NIC-N87 cell cultures for 48 hours under light, no light, and equal concentrations of astrodin as controls. Cell proliferation and viability were assessed by cell counting and other methods. Experimental data are shown in Table 1: Inhibition rate of MCF-7 tumor cells under no light / light for 150 mins and under no light / astrodin; Table 2: Inhibition rate of NIC-N87 tumor cells under no light / light for 150 mins and under no light / astrodin. This experiment demonstrates that the photocage compounds of this structure have inhibitory properties against the cytotoxicity of the original drug, with an inhibition factor of 15-193 times. The tumor cell inhibition rate under no light / light conditions clearly shows that light exposure increases the cytotoxicity of this photocage drug.

[0204] Table 1: Inhibition rate of MCF-7 tumor cells by non-light / light 150 mins and non-light / stellate aldehyde.

[0205] MCF-7(48H) 150 mins without light / with light Non-light / Astralaldehyde 150 mins of light exposure / stellaldehyde YB1 2.44 41.52 17.00 YB7 5.11 60.94 11.92 YB10 6.69 147.74 22.09 YB17 4.64 86.31 18.59 YB25 3.31 95.87 28.93 YB30 7.33 193.36 26.38

[0206] Table 2: Inhibition rate of NIC-N87 tumor cells by non-light / light 150 mins and non-light / stellate aldehyde.

[0207] NIC-N87(72H) 150 mins without light / with light Non-light / Astralaldehyde 150 mins of light exposure / stellaldehyde YB1 2.77 15.71 5.68 YB7 8.30 21.98 2.65 YB10 5.16 28.21 5.46 YB17 9.51 24.41 2.57 YB25 3.59 28.79 8.01 YB30 7.25 31.94 4.40

[0208] Mouse subcutaneous xenograft imaging

[0209] The compounds prepared in Examples 1-33 above were examined for imaging of subcutaneous xenografts in mice, and the fluorescence intensity of the de-cageing drugs in the tumors was monitored. The results showed that the compounds of the present invention can penetrate deep tissues in vivo through the photo-de-cageing process, as illustrated below.

[0210] Experimental methods:

[0211] Female BALB / c mice (20-25g) were inoculated with 4T1 cells (1×10⁻⁶) in their mammary fat pads. 6 (cells / mouse). When the tumor grows to approximately 200 mm... 3 Mice were randomly divided into a dark room group (unlit room) and a 660nm laser irradiation group (light irradiation group). Mice were anesthetized with isoflurane, and 10 μL of 0.1mM and 10mM CY5Q2 solutions were injected intratumorally. Subsequently, mice were either kept in darkness or irradiated with a 660nm laser (150mW / cm²). 2 The duration of the imaging varied from 0 to 40 minutes. Mice were imaged at 640 nm using a multimodal animal in vivo imaging system (BLT, AniView 600) with an excitation bandpass filter and an emission wavelength of 670 nm to monitor the fluorescence intensity of the anti-cage drug in the tumor.

[0212] Example 40

[0213] Continuous activation of CY5Q2 injected in situ enabled observation of the photocage-breaking process in live mice. After injecting CY5Q2 (0.1, 10 μM) into the tissue of female BALB / c mice (20-25g) at a depth of approximately 4 mm, specific fluorescence accumulation in the mice was detected after 660 nm laser irradiation, compared to the initial dark conditions, using a multimodal in vivo animal imaging system. The fluorescence intensity at the injection site showed a time-dependent increase, reaching a stable level after 25 minutes, consistent with the in vitro results. Notably, these results indicate that CY5Q2 can penetrate deep tissues through the photocage-breaking process to obtain in vivo feedback information.

[0214] Example 41

[0215] 4T1 cells (1×10⁻⁶) were inoculated onto the mammary fat pads of female BALB / c mice (20-25 g).6 (Cells / mouse). When the tumor grew to approximately 200 mm³, mice were randomly assigned to a dark group and a 660 nm laser irradiation group (light group). 10 μL of a 1.0 mM CYHR1 solution was injected intratumorally into each mouse. Subsequently, the mice were placed under either darkness or 660 nm laser irradiation (150 mW / cm²) for durations ranging from 0 to 9 minutes. CYHR1 levels in the tumor were monitored using a multimodal in vivo animal imaging system (BLT, AniView 600, China). After anesthesia with isoflurane, imaging was performed using an excitation bandpass filter at 640 nm and an emission bandpass filter at 670 nm.

[0216] Figure 7 This is an imaging image of a subcutaneous xenograft tumor in a CY5Q2 mouse. Figure 8 This is an imaging image of a subcutaneous xenograft tumor in a CYHR1 mouse. Figure 9 The images show fluorescence intensity curves of subcutaneous xenografts in CY5Q2(A) and CYHR1(B) mice.

Claims

1. An aldehyde hydrazone photocage compound having the following structural formula (I): ###0001### wherein Z is selected from C or N; W is selected from C or N; R2 is selected from H, halogen, nitro, C1-C6 alkyl, COOR3, OR4, SO2R5 or cyano; R3 is H or C1-C6 alkyl; R4 is H, benzyl, C1-C6 alkyl or halogen substituted C1-C6 alkyl; R5 is H, C1-C6 alkyl or amino; R1 is selected from the following groups: ###0002### A is an anion.

2. The compound of claim 1, wherein Z is C and W is selected from C or N.

3. The compound of claim 1, wherein Z is N and W is selected from C or N.

4. The compound of claim 1, having the following structural formula: ###0003### 5. The compound of claim 1, wherein said R2 is selected from H, fluorine, chlorine, bromine, iodine, nitro, cyano, methyl, ethyl, propyl, isopropyl, benzyloxy, -COOH, -COOCH3, -COOCH2CH3, -OH, -OCH3, -OCH2CH3, -OCF3, -SO2CH3 or -SO2NH2.

7. The compound of claim 1 is selected from the following specific compounds: ###0004### 2. The compound of claim 1, wherein 8. A method for preparing the aldehyde hydrazone photocage compound of any one of claims 1-7, by condensation reaction of formula (III) and formula (IV) to obtain the compound of formula (I), the reaction formula is as follows: ###0005### 3. The compound of claim 1, wherein 9. Use of the aldehyde hydrazone photocage compound of any one of claims 1-7 in the preparation of an antitumor drug.

4. The compound of claim 1, wherein 10. Use of the aldehyde hydrazone photocage compound of any one of claims 1-7 in the preparation of a photodynamic drug.

5. The compound of any one of claims 1-4, wherein the anion is chloride, bromide, iodide, sulfate, sulfite, nitrate, nitrite, phosphate, hydrogen phosphate, acetate, propionate, cinnamate, benzenesulfonate, citrate, lactate, or gluconate; the halogen is fluorine, chlorine, bromine, or iodine; the C 1-6 alkyl is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, or isohexyl; the halogen-substituted C 1-6 alkyl is selected from the group consisting of difluoromethyl or trifluoromethyl.

6. The compound of any one of claims 1-4, wherein ​ ​ ​ ​ ​

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