Chiral glucose modified zinc phthalocyanine as well as preparation method and application thereof

Abstract

The invention discloses chiral glucose modified zinc phthalocyanine as well as a preparation method and application thereof, solves the problem that a photosensitizer is low in enrichment degree at a tumor site, provides glucose small molecules with relatively good biocompatibility, modifies zinc phthalocyanine by using the glucose small molecules, and can be used for preparing the chiral glucose modified zinc phthalocyanine by adjusting a small molecule structure and chirality. Precise target tumor targeting of phthalocyanine is achieved, and then the in-vivo anti-tumor effect is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to a chiral glucose-modified zinc phthalocyanine and a preparation method and application thereof. Background Art

[0002] Photodynamic therapy (PDT) is an emerging platform with specific spatiotemporal selectivity and minimal invasiveness, and it has become an emerging solution for cancer treatment. PDT usually consists of three parts: photosensitizer (PS), light, and tissue oxygen. Irradiating the lesion with a specific wavelength can activate the photosensitizer that selectively accumulates in the lesion tissue, triggering a photochemical reaction to destroy the lesion. The photoactivated PS transfers its excited state energy to the surrounding oxygen to produce reactive oxygen species (ROS), which directly or indirectly cause cancer cell death by inducing tumor vascular stagnation. Similar to other therapies, the efficacy of PDT is limited by the tumor tissue enrichment and retention ability of photosensitizers. Existing photosensitizers have insufficient tumor targeting selectivity and limited retention ability in tumors. Therefore, the improvement of photosensitizer tumor targeting and retention ability is of great significance to the promotion and application of PDT in tumor treatment.

[0003] Glucose (Glu) is a monosaccharide widely distributed in nature and belongs to the class of polyhydroxyaldehyde compounds. In the body, glucose is mainly oxidized into water and carbon dioxide through the citric acid cycle to provide energy for the body, and plays a key role in the biosynthesis of phospholipid components such as ATP, DNA, and RNA. Glucose in the body is mainly D-Glu, which exhibits right-handed optical rotation. Among the different targeting methods, one targeting method is the glycosylation-mediated strategy. Due to the increase in the corresponding sugar receptors on the surface of cancer cells, the rate at which glucose enters tumor cells is 30 times higher than that of normal cells. Therefore, glucose has good tumor targeting specificity and good biocompatibility, while phthalocyanine is also facing the problem of insufficient tumor targeting ability in photodynamic therapy. Summary of the invention

[0004] To solve the above problems, the present invention discloses a chiral glucose-modified zinc phthalocyanine and a preparation method and application thereof, which solves the problem of low enrichment of photosensitizers at tumor sites, provides glucose small molecules with good biocompatibility and uses them to modify zinc phthalocyanine, and achieves precise targeting of phthalocyanine to target tumors by adjusting the structure and chirality of the small molecules, thereby enhancing the anti-tumor effect in vivo.

[0005] To achieve the above object, the technical solution of the present invention is as follows: A chiral glucose-modified zinc phthalocyanine, wherein the chiral glucose-modified zinc phthalocyanine has the following general structural formula: ; Wherein, R is selected from any of the following: , .

[0006] The present invention also provides a method for preparing the chiral glucose-modified zinc phthalocyanine: (1) When R = The following steps are included: Dissolve ZnPc-COOH4, L-Glu, TBTU and DIPEA in DMF and stir in an ice-water bath; after the reaction, drop the mixture into a beaker containing water and let it stand until the product is deposited at the bottom of the beaker. Collect the solid by centrifugation and wash the product with distilled water for several times; after vacuum drying, obtain the product L-Glu-ZnPc; The reaction formula is as follows: ; (2) When R = The following steps are included: Dissolve ZnPc-COOH4, D-Glu, TBTU and DIPEA in DMF and stir in an ice-water bath; after the reaction, drop the mixture into a beaker containing water and let it stand until the product is deposited at the bottom of the beaker. Collect the solid by centrifugation and wash the product with distilled water for several times; after vacuum drying, obtain the product D-Glu-ZnPc; The reaction formula is as follows:

[0007] As an improvement of the present invention, the molar ratio of ZnPc-COOH4, L-Glu, TBTU and DIPEA in the preparation method (1) is 1:4-20:4-20:10-100.

[0008] As an improvement of the present invention, the molar ratio of ZnPc-COOH4, D-Glu, TBTU and DIPEA in the preparation method (2) is 1:4-20:4-20:10-100.

[0009] As an improvement of the present invention, the stirring time in the ice-water bath in the preparation methods (1) and (2) is 6 to 24 hours.

[0010] Preferably, taking the preparation of L-Glu-ZnPc as an example, the preparation method (1) comprises the following steps: S1. Product synthesis: ZnPc-COOH4, L-Glu, TBTU and DIPEA were dissolved in DMF, the molar ratio of ZnPc-COOH4, L-Glu, TBTU and DIPEA was 1:4-20:4-20:10-100; the mixture was stirred in an ice-water bath for 6-24 hours; S2. Product purification: The obtained coupling product is poured into double distilled water to precipitate the product, and the precipitated solid is washed with water for multiple times and vacuum dried to obtain L-Glu-ZnPc.

[0011] The present invention also provides an application of chiral glucose-modified zinc phthalocyanine in the preparation of photodynamic drugs.

[0012] The chiral glucose-modified zinc phthalocyanine is used as a photosensitizer in the preparation of photodynamic drugs.

[0013] The present invention uses different chiral drugs with different activities on cells. After glucose is modified, phthalocyanine has better biocompatibility. In vivo anti-tumor research experiments show that phthalocyanine tumor targeting function becomes better. The present invention uses chiral molecules to modify photosensitive drugs, and regulates their enrichment and retention capabilities in tumor tissues by regulating the chirality of glucose.

[0014] The beneficial effects of the present invention are: (1) The chiral glucose-modified zinc phthalocyanine of the present invention has good tumor tissue selective enrichment ability; (2) The chiral glucose-modified zinc phthalocyanine of the present invention has the advantages of simple preparation method, abundant raw materials and low cost. The obtained derivatives can be used as photosensitizers and are widely used in the development of photodynamic drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The synthetic flow chart of L-Glu-ZnPc and D-Glu-ZnPc.

[0016] Figure 2 The structural diagrams of L-Glu-ZnPc and D-Glu-ZnPc.

[0017] Figure 3 : The hydrogen spectra of L-Glu-ZnPc and D-Glu-ZnPc.

[0018] Figure 4 The mass spectra of L-Glu-ZnPc and D-Glu-ZnPc.

[0019] Figure 5 Figure 1 is a comparison of L-Glu-ZnPc and D-Glu-ZnPc administered via tail vein, where: Figure A is the distribution of L-Glu-ZnPc in various major tissues and tumors in the body 24 hours after tail vein administration; Figure B is the distribution of L-Glu-ZnPc in various major tissues and tumors in the body 48 hours after tail vein administration; Figure C is an intensity bar graph of the distribution of L-Glu-ZnPc in various major tissues and tumors in the body 24 hours after intravenous administration.

[0020] Figure 6The figures are comparisons of tumor volumes in subcutaneous transplanted tumor model animals in mice after 14 days of treatment, where: Figure A is a comparison of tumor volume Vt / V0 values ​​(***P< 0.001, **P< 0.01: comparison between different types of drugs and light treatment groups and control groups; ###P<0.001, ##P<0.01: comparison between D-Glu modified phthalocyanine and L-Glu modified phthalocyanine); Figure B is a comparison of tumor volume; Figure C is a comparison of tumor volume on day 14 between D-Glu-ZnPc, L-Glu-ZnPc and control groups. DETAILED DESCRIPTION

[0021] The present invention will be further explained below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the following specific embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0022] The synthesis method of ZnPc-COOH4 is based on Arginine-Substituted Phthalocyanine with Concentration-Driven Self-Disaggregation Performance: Synthesis, Properties and Mechanistic Study. Asian Journal of Chemistry, 2016, 11: 3008-3013.

[0023] Example 1 Dissolve ZnPc-COOH4 (0.05 mmol), L-Glu (0.4 mmol), TBTU (0.4 mmol) and DIPEA (3 mmol) in DMF (4 mL) and stir in an ice-water bath for 12 h. After the reaction, add the mixture dropwise into a beaker containing 150 mL of water (50 mL / min) and let stand for 1 h. The product will settle at the bottom of the beaker. Collect the solid by centrifugation and wash the product 10 times with distilled water. After vacuum drying, the product L-Glu-ZnPc is obtained. The flow chart of the synthesis of L-Glu-ZnPc is shown in the figure. Figure 1 shown.

[0024] 1 H NMR (400 MHz, DMSO) δ 8.74 (d, J = 32.9 Hz, 1H), 8.31 (d, J = 33.7Hz, 2H), 7.84 – 7.44 (m, 5H), 3.08 (d, J = 17.6 Hz, 4H), 2.89 (s, 1H), 2.74(s, 1H). MS-MALDI-TOF (m / z): calcd for C84 H 72 N8O 32 Zn, 1768.35; found [M + H] + ,1769.53.

[0025] Example 2 Dissolve ZnPc-COOH4 (0.05 mmol), D-Glu (0.4 mmol), TBTU (0.4 mmol) and DIPEA (3 mmol) in DMF (4 mL) and stir in an ice-water bath for 12 h. After the reaction, add the mixture dropwise into a beaker containing 150 mL of water (50 mL / min) and let stand for 1 h. The product will settle at the bottom of the beaker. Collect the solid by centrifugation and wash the product 10 times with distilled water. After vacuum drying, the product D-Glu-ZnPc is obtained. The flow chart of the synthesis of D-Glu-ZnPc is shown in the figure. Figure 1 shown.

[0026] 1 H NMR (400 MHz, DMSO) δ 8.74 (d, J = 30.5 Hz, 1H), 8.48 – 8.11 (m,2H), 7.87 – 7.41 (m, 5H), 3.08 (d, J = 17.2 Hz, 4H), 2.89 (s, 1H), 2.74 (s,1H). MS-MALDI-TOF (m / z): calcd for C 84 H 72 N8O 32 Zn, 1768.35; found [M + H] + ,1769.81.

[0027] The structures of L-Glu-ZnPc and D-Glu-ZnPc prepared in Example 1-2 of the present invention are shown in FIG. Figure 2 As shown, the hydrogen spectrum and mass spectrum are Figure 3 As shown in Figure 4, it shows that the above two materials were successfully synthesized.

[0028] Example 3 Drug distribution in vivo, tumor accumulation and tumor tissue retention capacity detection When treating female Balb / c mice bearing tumors, experiments were performed using mice cultured with 4T1 cells when the tumors grew to 100 mm³. The mice were randomly divided into three groups and received tail vein injections of PBS (200 μL, pH=7.4) and two different ZnPcs (at a dose of 5 mg / kg mouse body weight). The two ZnPcs were the final products prepared by Example 1 and Example 2 and dissolved in 0.01 M DMSO to ensure that a dose of 5 mg / kg was injected into each mouse. During the experiment, the body weight and tumor volume of the mice were recorded every day. After a certain period of time (24 or 48 hours), the mice were euthanized and dissected, and the heart, liver, spleen, lungs, kidneys, brain and tumor tissues were removed. The fluorescence intensity of phthalocyanine in each organ and tumor was detected using a small animal fluorescence imaging device. The test results are as follows Figure 5 As shown in the figure, at 24 h, D-Glu-ZnPc and L-Glu-ZnPc were only enriched in the tumor site, indicating that both phthalocyanines have certain tumor targeting functions, but the targeting effect of L-Glu-ZnPc is significantly better than that of D-Glu-ZnPc. It was also found that after 48 h, D-Glu-ZnPc and L-Glu-ZnPc were almost completely metabolized in various organs and tumors, indicating that the metabolism of the phthalocyanine is fast, but the retention ability of tumor tissue is also poor.

[0029] Example 4 In vivo anticancer activity assay Using the method of Example 3, when the tumor volume reached 100 mm³, the tumor-bearing mice were divided into three groups and illuminated every day, and their body weight and tumor volume were measured. On days 1, 4, 7, 10 and 13, the mice were injected with PBS (200 μL, pH=7.4) and two ZnPcs synthesized in Example 1 and Example 2: D-Glu-ZnPc and L-Glu-ZnPc (dose of 3.2 mg / kg mouse body weight) through the tail vein. The final products of the two ZnPcs were dissolved in 0.01 M DMSO to ensure that each mouse received a dose of 3.2 mg / kg. Four hours after injection, the tumor tissue was irradiated with a 665 nm LED (power of 0.9 W / cm²) for 1 hour. After 14 days of treatment, the mice were euthanized, and the changes in tumor volume of each group of mice during the treatment period were recorded. As Figure 6 As shown, during the 14-day PDT treatment period, compared with the control group, both groups of phthalocyanines had tumor proliferation inhibitory effects, and the in vivo antitumor activity of L-Glu-ZnPc was stronger than that of D-Glu-ZnPc.

[0030] It should be noted that the above are only preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. For ordinary technicians in this technical field, several improvements and modifications can be made on the basis of the above embodiments without departing from the principles of the present invention. These improvements and modifications all fall within the scope of protection of the claims of the present invention.

Claims

1. A chiral glucose-modified zinc phthalocyanine, characterized in that: The general structural formula of the chiral glucose-modified zinc phthalocyanine is as follows: ； Wherein, R is selected from any of the following: 、 。 2. A method for preparing chiral glucose-modified zinc phthalocyanine according to claim 1, characterized in that: (1) When R = The following steps are included: Dissolve ZnPc-COOH4, L-Glu, TBTU and DIPEA in DMF and stir in an ice-water bath; after the reaction, drop the mixture into a beaker containing water and let it stand until the product is deposited at the bottom of the beaker. Collect the solid by centrifugation and wash the product with distilled water for several times; after vacuum drying, obtain the product L-Glu-ZnPc; (2) When R = The following steps are included: Dissolve ZnPc-COOH4, D-Glu, TBTU and DIPEA in DMF and stir in an ice-water bath. After the reaction is completed, add the mixture dropwise into a beaker filled with water and let it stand. The product will settle at the bottom of the beaker. Collect the solid by centrifugation and wash the product several times with distilled water. After vacuum drying, the product D-Glu-ZnPc is obtained.

3. The method for preparing chiral glucose-modified zinc phthalocyanine according to claim 2, characterized in that: In the preparation method (1), the molar ratio of ZnPc-COOH4, L-Glu, TBTU and DIPEA is 1:4-20:4-20:10-100.

4. The method for preparing chiral glucose-modified zinc phthalocyanine according to claim 2, characterized in that: In the preparation method (2), the molar ratio of ZnPc-COOH4, D-Glu, TBTU and DIPEA is 1:4-20:4-20:10-100.

5. The method for preparing chiral glucose-modified zinc phthalocyanine according to claim 2, characterized in that: The stirring time in the ice-water bath in the preparation methods (1) and (2) is 6 to 24 hours.

6. Use of the chiral glucose-modified zinc phthalocyanine according to claim 1 in the preparation of photodynamic drugs.

7. The use of chiral glucose-modified zinc phthalocyanine in fluorescent plastics according to claim 6, characterized in that: The chiral glucose-modified zinc phthalocyanine is used as a photosensitizer in the preparation of photodynamic drugs.

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