Pantothenic acid modified aza-BODIPY photosensitizer as well as synthesis method and application thereof
By introducing pantothenic acid groups into the aza-BODIPY photosensitizer, a photosensitizer with targeting capabilities was synthesized, solving the problem of aza-BODIPY photosensitizer accumulation in cancer cells and achieving highly efficient photodynamic therapy.
Patent Information
- Application Number
- CN202511206767.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-27
- Publication Date
- 2025-12-05
AI Technical Summary
The aza-BODIPY photosensitizer lacks targeted modification in photodynamic therapy, resulting in poor efficacy and safety issues, and it fails to effectively accumulate in cancer cells.
By introducing pantothenic acid groups into aza-BODIPY, pantothenic acid-modified aza-BODIPY photosensitizers are synthesized. A specific chemical synthesis route is used to link pantothenic acid derivatives with aza-BODIPY to form photosensitizers with targeting capabilities.
The synthesized pantothenic acid-modified aza-BODIPY photosensitizer exhibits good photosensitizing activity and cancer cell-specific uptake ability, with improved singlet oxygen production, making it suitable for photodynamic therapy of nonspecific and widespread tumors.
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Figure CN121064231A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of pharmaceutical synthesis technology, and relates to a pantothenic acid-modified aza-BODIPY photosensitizer, its synthesis method, and its uses. Background Technology
[0002] Photodynamic therapy (PDT) is a non-invasive technique that has emerged in the last 30 years, utilizing the photodynamic effect for the diagnosis and treatment of diseases. PDT offers advantages such as high precision, non-invasiveness, controllability, low toxicity, repeatability, and low likelihood of drug resistance, and has therefore been used clinically to treat some tumors and skin diseases. Photosensitizers are one of the key elements for exerting the photodynamic killing effect. Aza-BODIPY is a class of BODIPY dyes in which the carbon atom at the meso position is replaced by a nitrogen atom. These BODIPY dyes possess high molar absorptivity, desirable stability, and near-infrared absorption wavelengths. Therefore, aza-BODIPY is a more suitable molecular framework for modification into photosensitizers.
[0003] Studies show that introducing heavy atoms into aza-BODIPY effectively increases its singlet oxygen yield, thereby improving photodynamic therapy efficacy. Furthermore, introducing an iodine atom at position 6 of aza-BODIPY results in a high singlet oxygen yield (Φ... Δ =0.51, DMF) and a photosensitizer with good stability (compound 9).
[0004]
[0005] However, the application of aza-BODIPY photosensitizers in photodynamic therapy remains highly limited due to the lack of relatively effective targeting strategies. Untargeted photosensitizers may lead to poor photodynamic efficacy and safety issues; therefore, improving the effective accumulation of aza-BODIPY photosensitizers at the disease site through targeted modification is particularly important.
[0006] The myelocytomatosis oncogene (MYC) is a core transcription factor regulating cell proliferation. Elevated MYC protein levels in tumors are often associated with poor clinical outcomes in cancer patients. Cells with high MYC expression levels significantly increase their uptake of vitamin B5 (pantothenic acid), a finding that may provide a method for targeting cancer cells through metabolism. Therefore, introducing pantothenic acid into aza-BODIPY via chemical synthesis could potentially achieve targeted enrichment of photosensitizers in cancer cells. However, this targeted modification approach has not yet been used in the development of aza-BODIPY photosensitizers. Summary of the Invention
[0007] This invention addresses the technical problems of aza-BODIPY photosensitizers in photodynamic therapy by providing a pantothenic acid-modified aza-BODIPY photosensitizer, which has good photosensitizing activity and cancer cell-specific uptake ability. It can be further prepared into a targeted photodynamic drug for photodynamic therapy of non-specific, broad-spectrum tumors.
[0008] To achieve the above objectives, the present invention adopts the following technical solution:
[0009] In a first aspect, the present invention provides a pantothenic acid-modified aza-BODIPY photosensitizer, the general structural formula of which is shown in Formula IV:
[0010]
[0011] Where: R1 is selected from One of them, n = 1 to 6; the dashed line indicates whether the C-C bond exists or not.
[0012] Preferably, when n=1, R1 is At that time, the aza-BODIPY photosensitizer has the following structure:
[0013] The dashed line indicates whether the CC bond exists or not.
[0014] Preferably, when n=1, R1 is At that time, the aza-BODIPY photosensitizer has the following structure:
[0015] The dashed line indicates whether the CC bond exists or not.
[0016] Preferably, when n=1, R1 is At that time, the aza-BODIPY photosensitizer has the following structure:
[0017] The dashed line indicates whether the CC bond exists or not.
[0018] Preferably, when n=1, R1 is At that time, the aza-BODIPY photosensitizer has the following structure:
[0019] The dashed line indicates whether the CC bond exists or not.
[0020] Secondly, the present invention provides a method for synthesizing a pantothenic acid-modified aza-BODIPY photosensitizer, comprising the following steps:
[0021] 1) Precursor 1, as shown in Formula I, was dissolved in N,N-dimethylformamide (DMF), and potassium carbonate and a Boc-protected brominated reagent (Br-R1-Boc) were slowly added. The reaction was carried out at 90 °C for 1 h. After the reaction was completed, the mixture was extracted with water and dichloromethane, the organic phase was collected and dried, and the solvent was removed by rotary evaporation to obtain precursor 2, as shown in Formula II.
[0022] 2) Precursor 2 was dissolved in a mixed solvent of dichloromethane (DCM) and trifluoroacetic acid (TFA) and reacted at room temperature for 3 h; then the pH of the solution was adjusted to 5, and the solid was recrystallized in petroleum ether and dichloromethane and collected; then the solid was dissolved in a mixed solution of dichloromethane and acetic acid, N-iodosuccinimide (NIS) was added, and the reaction was carried out at room temperature for 2 h; after the reaction was completed, the solvent was evaporated to obtain precursor 3 as shown in Formula III;
[0023] 3) Precursor 3 was dissolved in dichloromethane, and then pantothenic acid derivative, N,N-diisopropylethylamine (DIPEA) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) were slowly added. The reaction was carried out at room temperature for 2 h. After the reaction was completed, the mixture was extracted with water and dichloromethane, the organic phase was collected and dried, the solvent was removed by rotary evaporation, and the final product shown in Formula IV was obtained by silica gel column chromatography.
[0024] The reaction pathway described above is as follows:
[0025]
[0026] Where: R1 is selected from One of them, n = 1 to 6; the dashed line indicates whether the C-C bond exists or not.
[0027] Preferably, the molar ratio of precursor 1 to the Boc-protected bromine reagent in step 1) is 1:2.
[0028] Preferably, in step 2), the volume ratio of dichloromethane to trifluoroacetic acid in the mixed solvent of dichloromethane and trifluoroacetic acid is 1:1, and the volume ratio of dichloromethane to acetic acid in the mixed solution of dichloromethane and acetic acid is 3:1.
[0029] Preferably, the molar ratio of precursor 2 to N-iodosuccinimide in step 2) is 1:1.1.
[0030] Preferably, the molar ratio of precursor 3, pantothenic acid derivative, DIPEA and HATU in step 3) is 1:1.5:2:2.
[0031] Thirdly, the present invention also provides the use of the pantothenic acid-modified aza-BODIPY photosensitizer in the preparation of targeted photodynamic drugs for treating nonspecific, broad-spectrum tumors and bacterial and fungal infections.
[0032] Preferably, the nonspecific broad tumor is selected from liver cancer, lung cancer, stomach cancer, breast cancer, colon cancer, pancreatic cancer, cervical cancer, bladder cancer, ovarian cancer, skin cancer, prostate cancer, lymphoma, or glioma.
[0033] Compared with the prior art, the beneficial effects of the present invention are as follows:
[0034] The pantothenic acid-modified aza-BODIPY photosensitizers synthesized in this invention exhibit good photosensitizing activity and cancer cell-specific uptake ability, such as the singlet oxygen yield (Ф) of photosensitizer compounds 1-8. Δ The concentrations (0.61–0.69) are all higher than those of the reference compound, and can be further prepared into targeted photodynamic drugs for photodynamic therapy of nonspecific, broad-spectrum tumors. Attached Figure Description
[0035] Figure 1 This shows the uptake of photosensitizer compound 3 of the present invention in cancer cells. Detailed Implementation
[0036] The following examples are used to illustrate the present invention, but are not intended to limit the scope of protection of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art. The experimental methods in the following examples are conventional methods unless otherwise specified. The reagents in the following examples are commercially available unless otherwise specified.
[0037] Example 1: Synthesis of compounds 1-8
[0038]
[0039] Aza-BODIPY precursor 1 (1 mmol) was dissolved in 100 mL of N,N-dimethylformamide (DMF), and potassium carbonate (10 mmol) and Boc-protected brominated reagent (2 mmol) were slowly added. The reaction was carried out at 90 °C for 1 h. After the reaction was completed, the mixture was extracted with water and dichloromethane, the organic phase was collected and dried, and the solvent was removed by rotary evaporation to obtain precursor 2.
[0040] Precursor 2 was dissolved in 50 mL of a mixed solvent of dichloromethane and trifluoroacetic acid (v:v = 1:1) and reacted at room temperature for 3 h. The pH of the solution was then adjusted to 5, and the solution was recrystallized in petroleum ether and dichloromethane, and the solid was collected. This solid was then dissolved in 50 mL of a mixed solution of dichloromethane and acetic acid (v:v = 3:1), and N-iodosuccinimide (1.1 mmol) was added. The reaction was carried out at room temperature for 2 h. After the reaction was complete, the solvent was evaporated to obtain precursor 3.
[0041] The precursor was dissolved in 50 mL of dichloromethane, followed by the slow addition of pantothenic acid derivative (1.5 mmol), DIPEA (2 mmol), and HATU (2 mmol), and the reaction was carried out at room temperature for 2 h. The mixture was extracted with water and dichloromethane, and the organic phase was collected and dried. After removing the solvent by rotation, the final product was obtained by silica gel column chromatography.
[0042] Photosensitizer compounds 1–8 were synthesized according to the above method, with yields of 47%, 27%, 39%, 66%, 42%, 51%, 29%, and 53%, respectively.
[0043] The structural formula of compound 1 is as follows:
[0044]
[0045] The 1H NMR spectrum data of compound 1 are as follows: 1 H NMR(600MHz, CDCl3)δ9.18(s,1H),8.09-7.99(m,5H),7.86-7.80(m,2H),7.72(d, J=8.6Hz,2H),7.50-7.42(m,3H),7.38(dd,J=5.2,1.9Hz,3H),7.10(s,1H),7.04-6 .98(m,2H),6.98-6.92(m,2H),5.85(s,1H),5.48(s,1H),4.24(s,2H),3.88(s,3H) ,3.48-3.45(m,4H),2.66(m,2H),0.89(s,6H).HRMS(ESI)(m / z):[M+H]+Calcd.for C43H41BF2IN5O6:Expected 899.22; found 899.22.
[0046] The structural formula of compound 2 is as follows:
[0047]
[0048] The 1H NMR spectrum data of compound 2 are as follows: 1H NMR (600MHz, CDCl3) δ8.09-7.99(m,4H),7.86-7.80(m,2H),7.72(d,J=8.6Hz,2H),7.50-7.42(m,3H),7.38(dd,J=5.2,1.9Hz,3H),7.10(s,1H),7.0 4-6.98(m,2H),6.98-6.92(m,2H),3.88-3.84(m,5H),3.58-3.48(m,4H),2 .05(m,1H),1.72(m,2H),1.47(m,2H).HRMS(ESI)(m / z):[M+H]+Calcd.for C48H49BF2IN5O6:Expected 967.28; found 967.28.
[0049] The structural formula of compound 3 is as follows:
[0050]
[0051] The 1H NMR spectrum data of compound 3 are as follows: H NMR (600MHz, CDCl3) δ8.09-7.99(m,4H),7.86-7.80(m,2H),7.72(d,J=8.6Hz,2H),7 .50-7.42(m,3H),7.38(dd,J=5.2,1.9Hz,3H),7.10(s,1H),7.04-6.98(m,2H),6.98 -6.92(m,2H),4.29(m,1H),4.04(m,1H),3.88(s,3H),3.54-3.46(m,4H),3.26(m,1H ),2.76-2.68(m,2H).HRMS(ESI)(m / z):[M+H]+Calcd.forC48H47BF2IN5O7:Expected 981.26; found 981.26.
[0052] The structural formula of compound 4 is as follows:
[0053]
[0054] The 1H NMR spectrum data of compound 4 are as follows: 1H NMR (600MHz, CDCl3) δ8.09-7.99(m,4H),7.86-7.80(m,2H),7.72(d,J=8.6Hz,2H),7 .50-7.42(m,3H),7.38(dd,J=5.2,1.9Hz,3H),7.10(s,1H),7.04-6.98(m,2H),6.98 -6.92(m,2H),4.01(m,1H),3.88(s,3H),3.78(m,1H),3.52-3.48(m,2H),3.42-3.24 (m,2H),2.44(m,1H),2.04(m,1H),1.78(m,1H).HRMS(ESI)(m / z):[M+H]+Calcd.for C47H47BF2IN5O6:Expected 953.26; found 953.26.
[0055] The structural formula of compound 5 is as follows:
[0056]
[0057] The 1H NMR spectrum data of compound 5 are as follows: 1 H NMR (600MHz, CDCl3) δ9.18 (s, 1H), 8.61 (d, J = 9.0Hz, 1H), 8.01 (s, 1H), 7.86-7.78 (m, 2H ),7.75-7.63(m,4H),7.53-7.34(m,6H),7.09-7.01(m,2H),6.91(dd,J=9.0,2.6Hz,1H), 6.82(d,J=2.6Hz,1H),5.85(s,2H),5.48(s,1H),4.24(s,2H),3.91(s,3H),3.48-3.45( m,4H),3.02-2.90(m,4H),2.66(m,2H),0.89(s,6H).HRMS(ESI)(m / z):[M+H]+Calcd.for C45H43BF2IN5O6:Expected 925.23; found925.23.
[0058] The structural formula of compound 6 is as follows:
[0059]
[0060] The 1H NMR spectrum data of compound 6 are as follows: 1H NMR (600MHz, CDCl3) δ8.61 (d, J = 9.0Hz, 1H), 8.01 (s, 1H), 7.86-7.78 (m, 2H), 7.75-7.63 (m, 4 H),7.53-7.34(m,6H),7.09-7.01(m,2H),6.91(dd,J=9.0,2.6Hz,1H),6.82(d,J=2.6Hz,1H), 5.48(s,1H),4.24(s,2H),3.91-3.88(m,5H),3.59-3.45(m,4H),3.02-2.90(m,8H),2.66(m, 2H),2.05(m,1H),1.72(m,2H),1.47(m,2H),0.89(s,6H).HRMS(ESI)(m / z):[M+H]+Calcd.for C50H51BF2IN5O6:Expected993.29; found 993.29.
[0061] The structural formula of compound 7 is as follows:
[0062]
[0063] The 1H NMR spectrum data of compound 7 are as follows: 1 H NMR (600MHz, CDCl3) δ8.61 (d, J = 9.0Hz, 1H), 7.86-7.78 (m, 2H), 7.75-7.63 (m, 4 H),7.53-7.34(m,6H),7.09-7.01(m,2H),6.91(dd,J=9.0,2.6Hz,1H),6.82(d,J =2.6Hz,1H),4.29(m,1H),4.04(m,1H),3.91(s,3H),3.54-3.46(m,4H),3.26(m ,1H),3.02-2.90(m,4H),2.76-2.66(m,2H).HRMS(ESI)(m / z):[M+H]+Calcd.for C50H49BF2IN5O7:Expected 1000.27; found 1000.27.
[0064] The structural formula of compound 8 is as follows:
[0065]
[0066] The 1H NMR spectrum data of compound 8 are as follows: 1H NMR (600MHz, CDCl3) δ8.61 (d, J = 9.0Hz, 1H), 7.86-7.78 (m, 2H), 7.75-7.63 (m, 4H), 7.5 3-7.34(m,6H),7.09-7.01(m,2H),6.91(dd,J=9.0,2.6Hz,1H),6.82(d,J=2.6Hz,1H), 3.98(m,1H),3.91(s,3H),3.73(m,1H),3.52-3.51(m,2H),3.41-3.27(m,2H),3.02-2. 90(m,4H),2.47(m,1H),2.04(m,1H),1.78(m,1H).HRMS(ESI)(m / z):[M+H]+Calcd.for C49H49BF2IN5O6:Expected 979.28; found 979.28.
[0067] Example 2 Singlet Oxygen Yield Test
[0068] The photosensitizers in this embodiment are compounds 1-8, compound 9 (see background art), and zinc phthalocyanine (ZnPc) prepared in Example 1.
[0069] 2 mL of a 100 μM DMF solution of 1,3-diphenylisobenzofuran (DPBF) was mixed thoroughly with 2 mL of a 2 μM DMF solution of the photosensitizer and placed in a quartz cuvette. The mixture was then exposed to light for different durations (0 s, 10 s, 20 s, 30 s, 40 s, 50 s, 60 s) using a 90 mW light source. The cutoff filter wavelength was 550 nm. The UV-Vis absorption spectra of the photosensitizer solution after different illumination durations (0 s, 10 s, 20 s, 30 s, 40 s, 50 s, 60 s) were measured in the wavelength range of 350–800 nm. The singlet oxygen yield of the photosensitizer was calculated based on the change in DPBF absorbance at 415 nm.
[0070] The results are shown in Table 1. The results indicate that the singlet oxygen yield (Φ) of the pantothenic acid-modified aza-BODIPY photosensitizer compounds 1-8 prepared in this invention is... Δ =0.61~0.69) are all higher than those of the unmodified aza-BODIPY photosensitizer compound 9 (Φ Δ =0.52) and the singlet oxygen yield of ZnPc (Φ Δ =0.56).
[0071] Table 1 Results of photosensitizer singlet oxygen yield test
[0072] photosensitizer 1 2 3 4 5 6 7 8 9 ZnPc <![CDATA[Φ Δ ]]> 0.64 0.67 0.68 0.69 0.64 0.61 0.64 0.68 0.52 0.56
[0073] Example 3: In vitro antitumor activity test
[0074] The in vitro antitumor activity of photosensitizers (compounds 1-8, compound 9, and zinc phthalocyanine (ZnPc)) was determined using the CCK-8 assay. 5000 HeLa cells were seeded per well in 96-well plates and cultured overnight at 37°C. After discarding the original culture medium, 100 μL of culture medium containing different concentrations (10 μM, 2.5 μM, 0.625 μM, 156.25 nM, 39.06 nM, and 9.77 nM) of photosensitizer were added to each well. The cells were incubated at 37°C for 3 h, followed by light treatment and then cultured for another 24 h. Each experiment was repeated three times. The light treatment time was 5 min, the light source power was 90 mW, and the cutoff filter wavelength was 550 nm. After removing the culture medium, DMEM medium containing 10% CCK-8 was added, and the cells were incubated at 37°C for another 2 h. The OD value at 450 nm was measured using a microplate reader. The IC50 value was calculated based on the microplate reader reading.
[0075] The results are shown in Table 2. The results indicate that the photosensitizer compounds 1–8 synthesized in this invention exhibit better phototoxicity than compound 9 and ZnPc. Among them, photosensitizer 3 exhibits the strongest phototoxicity, at 0.037 μM.
[0076] Table 2. Phototoxicity test results of photosensitizer on HeLa cells (IC50) 50 Value, unit: μM).
[0077] photosensitizer 1 2 3 4 5 6 7 8 9 ZnPc <![CDATA[IC 50 ]]> 0.061 0.054 0.037 0.052 0.048 0.049 0.074 0.055 0.285 3.761
[0078] Example 4 Cell Uptake Test
[0079] HeLa and HUVEC cells were cultured in DMEM medium (purchased from Beyotime Biotechnology) for 24 h. After washing three times with PBS, DMEM medium containing photosensitizer compound 3 (10 μM) was added, and the cells were incubated at 37°C for 0 h, 0.5 h, 1 h, 2 h, and 3 h, respectively. The original medium was discarded, and the cells were washed three times with PBS. The uptake of the photosensitizer in the cells was then observed using laser confocal microscopy. Results are as follows: Figure 1 As shown, photosensitizer compound 3 exhibits good uptake ability by cancer cells.
[0080] Therefore, the pantothenic acid-modified aza-BODIPY photosensitizer synthesized in this invention can be prepared as a novel photodynamic therapy drug targeting cancer cells.
[0081] The embodiments described above are merely preferred embodiments of the present invention and are only used to explain the present invention. They are not intended to limit the scope of the present invention. For those skilled in the art, other implementation methods can be easily made by substitution or modification based on the technical content disclosed in this specification. Therefore, all changes and improvements made on the principle of the present invention should be included within the scope of the patent application of the present invention.
Claims
1. A pantothenic acid-modified aza-BODIPY photosensitizer, characterized in that, The structural general formula is shown as formula IV: wherein: R1is selected from one of the following, n = 1-6; the dotted line indicates the presence or absence of a C-C bond.
2. The pantothenic acid-modified aza-BODIPY photosensitizer according to claim 1, characterized in that, When n = 1, R1is When n = 1, R1is When n = 1, R1is When n = 1, R1is When n Dashed lines indicate the presence or absence of a C-C bond.
3. The pantothenic acid-modified aza-BODIPY photosensitizer according to claim 1, characterized in that, When n = 1, R1is When n = 1, R1is When n = 1, R1is When n = 1, R1is When n Dashed lines indicate the presence or absence of a C-C bond.
4. The pantothenic acid-modified aza-BODIPY photosensitizer of claim 1, wherein When n = 1, R1is When n = 1, R1is When n = 1, R1is When n = 1, R1is When n Dashed lines indicate the presence or absence of a C-C bond.
5. The pantothenic acid-modified aza-BODIPY photosensitizer according to claim 1, wherein When n = 1, R1is when n = 1, R1is when n = 1, R1is Dashed lines indicate the presence or absence of a C-C bond.
6. A method of synthesis of pantothenic acid-modified aza-BODIPY photosensitizers according to any one of claims 1 to 5, characterized in that, The method comprises the following steps: 1) dissolving precursor 1 shown in formula I in DMF, slowly adding potassium carbonate and Boc-protected brominating reagent, and reacting at 90°C for 1 h; after the reaction is completed, extracting with water and dichloromethane, collecting the organic phase and drying, and removing the solvent to obtain precursor 2 shown in formula II; 2) dissolving precursor 2 in a mixed solvent of DCM and TFA, and reacting at room temperature for 3 h; then adjusting the pH value of the solution to 5, recrystallizing in petroleum ether and dichloromethane and collecting the solid; then dissolving the solid in a mixed solution of dichloromethane and acetic acid, adding NIS, and reacting at room temperature for 2 h; after the reaction is completed, removing the solvent to obtain precursor 3 shown in formula III; 3) dissolving precursor 3 in dichloromethane, then slowly adding a pantothenic acid derivative, DIPEA and HATU, and reacting at room temperature for 2 h; after the reaction is completed, extracting with water and dichloromethane, collecting and drying the organic phase, removing the solvent, and then performing silica gel column chromatography to obtain the final product shown in formula IV; The above reaction route is as follows: wherein: R1is selected from one of the following, n = 1-6; the dotted line indicates the presence or absence of a C-C bond.
7. Use of a pantothenic acid modified aza-BODIPY photosensitizer according to any one of claims 1 to 5 in the preparation of a targeted photodynamic drug for the treatment of non-specific widespread tumors and bacterial, fungal infections.
8. Use according to any one of claim 7, characterized in that, The non-specific widespread tumors are selected from liver cancer, lung cancer, gastric cancer, breast cancer, colon cancer, pancreatic cancer, cervical cancer, bladder cancer, ovarian cancer, skin cancer, prostate cancer, lymphoma or brain glioma.