Preparation method and application of parathyroid gland targeting cyanine dye photosensitizer

Abstract

The invention discloses a photosensitizer capable of targeting to parathyroid glands and a preparation method of the photosensitizer. A compound structure of the photosensitizer is shown as a formula I in the specification. The ICG dye is a commercial dye parent, through a series of screening on the ICG dye, targeting on parathyroid glands is expected to be achieved through the inherent targeting characteristics of the structure, meanwhile, modification is conducted on the two sides of the T800 cyanine dye, asymmetric F atoms, I atoms, F atoms and Br atoms are introduced, the existence of the F atoms guarantees the targeting efficiency, and meanwhile the targeting efficiency is improved. The introduction of the heavy atoms provides a photodynamic therapy effect, and precise targeting imaging and treatment of parathyroid glands are realized. In addition, contrast molecules with double F, double I and double Br introduced into symmetrical positions on two sides are synthesized for comparison. Results show that the T800-FI can realize accurate targeting on parathyroid glands, has an optimal photodynamic ablation effect on the parathyroid glands, can be applied to photodynamic therapy of diseases such as parathyroid carcinoma, parathyroid tumor, hyperparathyroidism and parathyroid cyst, and can realize the effect of accurate recognition and positioning therapy.

Description

Technical Field

[0001] The present invention belongs to the intersectional field of medicine and engineering, and specifically relates to a parathyroid targeted photosensitizer constructed based on heptamethine cyanine dye, which has good phototoxicity to cancer cells and parathyroid cells, and has good targeting and phototherapy effects on parathyroid tissue. Background Art

[0002] The parathyroid gland is one of the endocrine glands in the human body. It is the size of two pairs of mung beans. Its color and size are very similar to those of the thyroid gland, but its location varies from person to person. It is usually located in the middle and lower part of the back of the left and right lobes of the thyroid gland. Its main function is to secrete parathyroid hormone and regulate the metabolism of calcium and phosphorus in the body. Currently, there are two common types of parathyroid diseases. One is hyperparathyroidism caused by increased parathyroid hormone, including parathyroid adenoma, etc. The other is hypoparathyroidism caused by decreased parathyroid hormone. Symptoms include decreased blood calcium, increased blood phosphorus, and epileptic seizures. Both hyperparathyroidism and hypoparathyroidism can cause disease in people. The clinical manifestations are highly heterogeneous. In addition, the disease is relatively rare and clinical physicians have limited knowledge, which can easily lead to missed diagnosis and misdiagnosis.

[0003] For the treatment of parathyroid diseases, the location of the parathyroid gland varies from person to person and the size is small. Traditional surgical resection is prone to miscutting or incomplete cutting. Chemotherapy and radiotherapy also have limitations such as expensive drugs, large side effects, and long-term use. Therefore, new diagnostic and treatment methods with high recognition accuracy, low toxicity and side effects, and time-space tracking of disease progression are urgently needed. At present, many new treatment methods have been developed, such as radiofrequency therapy and microwave ablation therapy. Radiofrequency ablation is a tumor thermal therapy method. Its basic principle is to use heat energy to kill tumor cells, but radiofrequency therapy has the risk of postoperative recurrent laryngeal nerve paralysis and thyroid autoimmune diseases caused by the body's stress response. Microwave ablation therapy is currently a new treatment for microadenoma. It is favored due to its advantages such as less intraoperative bleeding and less trauma. However, when the power of microwave ablation therapy is high, it will cause blood supply damage to the parathyroid glands or parathyroid gland damage, and patients will experience numbness of the limbs, and even convulsions, muscle spasms and other manifestations of hypocalcemia.

[0004] Photodynamic therapy has a series of characteristics such as strong targeting, little organ damage, non-invasiveness, low toxicity and side effects, and repeatable treatment. It is currently commonly used in the treatment of skin diseases and cancer, and rarely used in the treatment of microglandular diseases. It has significant advantages in targeting microglandular diseases such as parathyroid glands. Photosensitizers often have disadvantages such as complex synthesis, short absorption and emission wavelengths, and poor biocompatibility. Cyanine dyes have the characteristics of easy modification, strong tissue penetration, and good biocompatibility, and can play their unique advantages in the treatment of parathyroid tumors.

[0005] The parathyroid glands are very small and their color is difficult to identify compared to the thyroid gland, making it very difficult to remove or ablate the parathyroid glands. Therefore, how to accurately identify the parathyroid glands is a key difficulty that we need to solve. Previous literature reported that the heptamethine cyanine dye modified with the F element has structural intrinsic targeting and can specifically target the parathyroid glands without interference from the thyroid gland. Therefore, we envision using this strategy to develop a photosensitizer that can maintain accurate labeling of the parathyroid glands and has a photodynamic therapy effect, which has important research significance for the treatment of parathyroid-related diseases. Summary of the invention

[0006] Purpose of the invention: ICG dye is a commercial dye matrix. Through a series of screening, we hope to use the inherent targeting characteristics of the structure to achieve targeting of the parathyroid glands. At the same time, we modify the two sides of the T800 cyanine dye and introduce asymmetric F, Br, and I atoms. While ensuring the targeting efficiency, the introduction of heavy atoms provides a photodynamic therapy effect, achieving precise targeted imaging and treatment of the parathyroid glands.

[0007] The technical solution of the present invention: the compound of formula I:

[0008]

[0009] Wherein, X1 and X2 are each independently F, Br or I, and X1 and X2 are not F or Br at the same time;

[0010] Y - is an anion, and the anion has the same charge number as the cation in formula I;

[0011] Y - F - , Cl - Br - ,I - 、BF4 - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - or CF3SO3 - .

[0012] Specifically, the compound is selected from the following structures:

[0013]

[0014] Specifically, the compound is selected from the following structures:

[0015]

[0016] Specifically, the compound is selected from the following structures:

[0017]

[0018] Specifically, Y - F - , Cl - , Br or I - .

[0019] Application of the above-mentioned compounds in the preparation of photodynamic therapy drugs

[0020] The above-mentioned compound is used for preparing medicine for treating cancer or parathyroid disease.

[0021] The present invention also provides a method for preparing the compound of formula I:

[0022] The synthesis steps are as follows:

[0023]

[0024] Compound A and pentadienaldehyde diphenylamine hydrochloride react in acetic anhydride to obtain compound B.

[0025] In an inert gas atmosphere, compound B and compound C are reacted in acetic anhydride in the presence of a catalyst at room temperature to obtain a compound of formula I; the catalyst is one of sodium acetate and pyridine.

[0026] Y- is an anion, and the anion has the same charge number as the cation in formula I.

[0027] Y - F - , Cl - Br - ,I - 、BF4 - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - or CF3SO3 - .

[0028] Specifically, the present invention also provides a method for preparing a parathyroid targeted photosensitizer T800-X1X2, the steps of which are as follows:

[0029]

[0030] (1) Compound 1 is dissolved in water and HCl, and sodium nitrite is added dropwise, and the mixture is stirred in an ice bath. Stannous chloride is then added dropwise, and the reaction is continued at room temperature; the solvent is removed and purified to obtain compound 2;

[0031] (2) Compound 2 and 3-methyl-2-butanone were dissolved in acetic acid, and the reaction mixture was refluxed under a N2 atmosphere. After the reaction was completed, the solvent was removed and purified to obtain compound 3;

[0032] (3) Compound 3 and iodomethane were dissolved in acetonitrile and refluxed under N2 atmosphere. After the reaction, the solvent was removed and purified to obtain compound 4;

[0033] (4) Compound 4 and pentadienaldehyde diphenylamine hydrochloride are added to acetic anhydride in a certain proportion. After the reaction is completed, sodium carbonate is added to adjust the mixture to neutrality, and the solvent is removed to obtain compound 5, which is directly used for the next step of reaction;

[0034] (5) The crude product 5 of the previous step and the compound 3 having a substituent of X2 are dissolved in acetic anhydride according to a certain ratio, a catalyst is added, and the mixture is stirred at room temperature under a N2 atmosphere. After the reaction is completed, the mixture is purified to obtain the photosensitizer T800-X1X2.

[0035] Furthermore, in the above technical solution, the halogen atom combination of X1 and X2 may be one of (F, Br), (F, I), (F, F), (I, I) and (Br, Br).

[0036] Furthermore, in the above technical solution, the catalyst is one of sodium acetate and pyridine.

[0037] Furthermore, in the above technical solution, the method for removing the solvent includes suction filtration and reduced pressure evaporation; the method for purification includes extraction and column chromatography.

[0038] Furthermore, in the above technical solution, real-time TLC monitoring is performed during the reaction of step (2) and step (4).

[0039] Furthermore, in the above technical scheme, in step (1), the reaction time of the reactants in an ice bath is 2 hours, and the reaction time at room temperature is 2 hours; in step (3), the reflux time of the reactants is 18 hours; and in step (5), the mixture is stirred at room temperature for 2 hours under a N2 atmosphere.

[0040] Finally, the present invention provides an application of a parathyroid targeted photosensitizer, which is applied to explore the effects of different halogen atom substituents on the phototoxicity of cyanine dye photosensitizers by testing the singlet oxygen yield of T800-X1X2; MCF-7 cells are incubated with a culture medium containing T800-X1X2 and applied to cell uptake imaging to explore the differences in the uptake efficiency of photosensitizers caused by different halogen atom substituents; 4T1 cells, HepG-2 cells and MCF-7 cells are incubated with a culture medium containing photosensitizer T800-X1X2 and illuminated, and are applied to photodynamic therapy of cancer cells; the photosensitizer T800-X1X2 is injected into the body through the tail vein, and the photosensitizer of each group is evaluated according to the fluorescence imaging results of the photosensitizer in the parathyroid gland part to determine whether the photosensitizer of each group targets the parathyroid gland and its ability to target the parathyroid gland; the parathyroid gland part of the rat is illuminated under the same conditions, and the photodynamic therapy effect of the photosensitizer of each group on the parathyroid gland of the rat is evaluated by measuring the decrease levels of parathyroid hormone and blood calcium concentration in the rat.

[0041] Furthermore, in the above technical solution, the numerical change of singlet oxygen of each group of photosensitizer T800-X1X2 under the same lighting conditions in the solution is determined as an evaluation index of the effect of different halogen atom substituents on the in vitro phototoxicity of the photosensitizer.

[0042] The detection process of the photosensitizer on singlet oxygen in the solution under light is as follows: each group of T800-X1X2 is dissolved in a dimethyl sulfoxide solution to prepare a 10 mM T800-X1X2 mother solution. Take 3 mL of dichloromethane in a quartz dish, add 1,3-diphenylisobenzofuran (DPBF) and T800-X1X2 mother liquor until the ultraviolet absorption intensity of DPBF is about 1 and the ultraviolet absorption intensity of T800-X1X2 is about 0.3, then carry out continuous illumination with an interval of 30 s and a total duration of 300 s under an 808 nm, 3 mW laser, and then measure the decrease in the ultraviolet absorption intensity of DPBF; then replace T800-X1X2 with ICG, repeat the above steps, and finally calculate the singlet oxygen yield of T800-X1X2 in each group based on the measured value of the decrease in the ultraviolet absorption intensity of DPBF, and use this as an evaluation index for the effect of different halogen atom substituents on the in vitro phototoxicity of T800-X1X2.

[0043] Furthermore, in the above technical solution, the cell imaging fluorescence intensity of each group of photosensitizer T800-X1X2 is measured as an evaluation index for the effect of different halogen atom substituents on the cellular uptake efficiency of the photosensitizer.

[0044] Among them, the detection process of the photosensitizer on the in vitro cell uptake efficiency is as follows: for in vitro cells, take 2μL of each group of T800-X1X2 mother solution with a concentration of 10mM, dissolve it in 2mL of culture medium, and prepare a 10μM incubation solution for standby. At this time, the pH value of the incubation solution is 7.4-7.5, and the temperature is 37°C; MCF-7 cells are incubated with a culture medium containing the same concentration of T800-X1X2, and the changes in the fluorescence intensity of cells incubated with each group of T800-X1X2 are measured over time, and this is used as an evaluation index for the effect of different halogen atom substituents on the cell uptake efficiency of the photosensitizer.

[0045] Furthermore, in the above technical solution, the cytotoxicity result of T800-X1X2 is measured to reflect the evaluation index of the effect of different halogen atom substituents on the cytotoxicity of the photosensitizer.

[0046] The detection process of the photosensitizer on in vitro cytotoxicity is as follows: for in vitro cells, take 10 μL of each group of T800-X1X2 mother solution with a concentration of 10 mM, dissolve it in 90 μL of dimethyl sulfoxide solution, and prepare a mother solution with a concentration of 1 mM for use. Take 0 μL, 0.5 μL, 1 μL, 1.5 μL, 2 μL, 3 μL, 4 μL, and 5 μL of the mother solution with a concentration of 1 mM, respectively, dissolve it in 1 mL of culture medium, and prepare 0 μM, 0.5 μM, 1 μM, 1.5 μM, 2 μM, 3 μM, 4 μM, and 5 μM incubation solutions for use. At this time, the pH value of the incubation solution was 7.4-7.5, and the temperature was 37°C; after incubating 4T1 cells, HepG-2 cells and MCF-7 cells with a culture medium containing T800-X1X2 with the same concentration gradient for 1 hour, they were continuously illuminated at 808nm, 100mW for 5 minutes, and then incubated at 37°C for 24 hours. The culture medium was dried with cotton, and then thiazolyl blue was added and incubated for 4 hours. After the culture medium was dried with cotton, dimethyl sulfoxide solution was added to test the absorbance of the solution at 490nm, and the survival rate of cells incubated with each group of photosensitizers was calculated, and this was used as an evaluation index for the effect of different halogen atom substituents on the cytotoxicity of each group of photosensitizers.

[0047] Furthermore, in the above technical scheme, whether the photosensitizer targets the parathyroid gland and its targeting ability are evaluated by measuring the fluorescence imaging results of T800-X1X2 in the parathyroid gland area, and the photodynamic therapy effects of different photosensitizers on the parathyroid gland of rats are evaluated by measuring the decrease levels of parathyroid hormone and blood calcium concentration in rats.

[0048] The detection process of the photosensitizer on parathyroid tissue targeting and therapeutic effect is as follows: a PBS solution containing T800-X1X2 is prepared for standby use, the concentration of T800-X1X2 in the PBS solution is 1mM and 5mM, the prepared PBS solution is injected into the body through the tail vein, and the change in fluorescence intensity at the parathyroid gland over time is measured, and this is used as an evaluation index for the effect of different halogen atom substituents on the parathyroid targeting of the photosensitizer; an 808nm laser is used, and continuous illumination is performed at 200mW for 10min, and blood is collected at 0h, 6h, 24h, 36h, 3d, 5d and 10d to measure the PTH level and blood calcium concentration, and this is used as an evaluation index for the effect of each group of photosensitizers on the phototherapy effect of the parathyroid gland.

[0049] The T800-X1X2 of the present invention uses the inherent targeting characteristics of the structure to achieve parathyroid targeting. The two sides of the T800 cyanine dye are easy to modify. The introduction of F atoms can greatly improve the targeting efficiency. At the same time, the introduction of heavy atoms gives T800 excellent photodynamic therapy effects. The two work together to achieve precise targeted imaging and treatment of the parathyroid glands. The photodynamic therapy effect of T800-X1X2 on the parathyroid glands of small animals is evaluated by detecting the fluorescence imaging results of T800-X1X2 in the parathyroid glands of small animals, and the decrease levels of parathyroid hormone and blood calcium concentration. T800-FI has high cell uptake efficiency and excellent parathyroid targeting while meeting certain photodynamic therapy effects.

[0050] The T800-X1X2 described in the present invention has the following outstanding advantages when used for the treatment of parathyroid tumors:

[0051] (1) Simple synthesis: T800-X1X2 targets the parathyroid glands by virtue of its inherent structural targeting property, without the need to introduce additional targeting groups, and is easy to synthesize;

[0052] (2) Good targeting: The F atom introduced in T800-X1X2 cooperates with the inherent targeting property of the cyanine dye structure, which can ensure its high targeting efficiency to the parathyroid gland;

[0053] (3) Fast targeting speed: Cyanine dyes have a fast metabolic rate and can quickly reach the vicinity of the parathyroid gland after injection through the tail vein;

[0054] (4) Good phototherapy effect: T800-X1X2 can cause powerful phototoxicity to both cancer cells and normal parathyroid cells, and has the best photodynamic ablation effect on parathyroid glands, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1It is the singlet oxygen yield test graph of T800-FI, T800-FBr, T800-FF, T800-BrBr and T800-II in solution;

[0056] Figure 2 It is the test picture of T800-FI, T800-FBr, T800-FF, T800-BrBr and T800-II cell uptake confocal fluorescence imaging;

[0057] Figure 3 It is the cytotoxicity test chart of T800-FI, T800-FBr, T800-BrBr and T800-II;

[0058] Figure 4 It is the time-dependent fluorescence imaging test diagram of T800-FBr and T800-FI in the parathyroid gland;

[0059] Figure 5 This is a test chart of blood calcium concentration in rats after T800-FBr and T800-FI photodynamic therapy;

[0060] Figure 6 This is the test graph of PTH level in rats after photodynamic therapy with T800-FBr and T800-FI;

[0061] Figure 7 This is the H-NMR spectrum of T800-FI.

[0062] Figure 8 This is the H-NMR spectrum of T800-FBr.

[0063] Fig. 9 This is the H NMR spectrum of T800-BrBr.

[0064] Fig.10 This is the H-NMR spectrum of T800-II.

[0065] Fig.11 This is the H-NMR spectrum of T800-FF. DETAILED DESCRIPTION

[0066] The technical solution of the present invention is further described below in conjunction with specific embodiments.

[0067] Embodiment 1:

[0068] In this example, the following five parathyroid-targeted photosensitizers were synthesized:

[0069]

[0070] The synthetic route taking T800-FBr as an example is:

[0071]

[0072] Prepared in the following manner:

[0073] (1) Compound 1 (6.000 g, 1 eq) was dissolved in 4 mL of water and 20 mL of 37% concentrated HCl (v:v = 1:5), stirred in an ice bath for 10 min, and then 10 mL of 20.8% sodium nitrite (2.085 g, 1.1 eq) solution was slowly added dropwise. The reaction was continued for 1 h until a brown-yellow clear solution was obtained, and then stannous chloride (10.421 g, 2 eq) was dissolved in 7 mL of water and 7 mL of 37% concentrated HCl (v:v = 1:1), stirred at room temperature for 2 h until the solution turned light pink, and filtered to obtain solid compound 2 (4.125 g, 68.7% yield); 1 HNMR (400MHz, DMSO-d6) δ7.45(d,J=8.5Hz,2H),7.04(d,J=16.0Hz,1H),6.92–6.80(m,4H),2.62(s,2H),2.48(s,2H),1.10(s,6H).

[0074] (2) Compound 2 (5.000 g, 1 eq) and 3-methyl-2-butanone (3.678 g, 2 eq) were placed in a 250 mL single-mouth bottle, 50 mL of acetic acid was added to dissolve, and the mixture was refluxed at 120° C. under a N2 atmosphere. The reaction was monitored by TLC spot plate. After completion, saturated sodium carbonate solution was added to remove acetic acid until no more bubbles were generated, and the mixture was extracted three times with saturated sodium chloride aqueous solution and ethyl acetate, dried over anhydrous sodium sulfate, and the organic phase was evaporated under reduced pressure. The crude product was purified by column chromatography (EA:PE=1:1) to obtain a brown liquid product, compound 3 (3.782 g, 43.5% yield); 1 H NMR (500MHz, DMSO-d6) δ8.27 (s, 1H), 7.98 (d, 1H, J = 5.0Hz), 7.70 (d, 1H, J = 5.0Hz), 3.92 (s, 3H), 2.72 (s, 3H), 1.50 (s, 6H); 13 CNMR(125MHz,DMSO-d6)δ196.8,144.9,142.6,138.7,132.9,117.8,96.8,55.0,35.5,22.5,15.3.

[0075] (3) Compound 3 (1.500 g, 1 eq) and iodomethane (2.694 g, 2 eq) were dissolved in 10 mL of acetonitrile and refluxed overnight under N2 atmosphere. Solid precipitation was observed the next day. After the reaction was completed, ether was added to the reaction mixture, and the mixture was ultrasonically filtered to obtain a brown solid product, Compound 4 (1.210 g, 28.6% yield); 1H NMR (500MHz, DMSO-d6) δ8.27 (s, 1H), 7.98 (d, 1H, J = 5.0Hz), 7.70 (d, 1H, J = 5.0Hz), 3.92 (s, 3H), 2.72 (s, 3H), 1.50 (s, 6H); 13 CNMR(125MHz,DMSO-d6)δ196.8,144.9,142.6,138.7,132.9,117.8,96.8,55.0,35.5,22.5,15.3.

[0076] (4) Compound 4 (0.500 g, 1 eq) and pentadienaldehyde diphenylamine hydrochloride (0.427 g, 1.1 eq) were placed in a 50 mL round-bottom flask, 10 mL of acetic anhydride was added, and the mixture was stirred at 120°C for 1 h in a N2 atmosphere in the dark, and the reaction was detected by TLC. After the reaction, saturated sodium carbonate solution was added to adjust the pH to neutral, and then the mixture was extracted three times with dichloromethane and saturated brine, dried over anhydrous sodium sulfate, and the organic phase was spin-dried to obtain a red solid crude product 5 (0.348 g, 37.5% yield), which was directly used for the next step of the reaction;

[0077] (5) The crude product 5 (0.348 g, 1 eq) and 3-Br (0.736 g, 1.1 eq) from the previous step were placed in a 50 mL round-bottom flask, and 10 mL of acetic anhydride and 1 mL of pyridine were added. The mixture was stirred at 25 °C under N2 atmosphere in the dark for 0.5 h, and the reaction was monitored by TLC. After the reaction, saturated sodium carbonate solution was added to adjust the pH to neutral, and then extracted with dichloromethane and saturated brine for 3 times, dried over anhydrous sodium sulfate, and the organic phase was spin-dried and purified by column chromatography (DCM: MeOH = 50: 1) to obtain a dark green solid T800-FBr (0.214 g, 19.7% yield); 1H-NMR (400MHz, DMSO-d6) δ: 1.71 (s, 12H), 3.60 (s, 6H), 6.28 (d, J = 13.6 Hz, 2H), 6.60 (t, J = 12.8 Hz, 2H), 7.26 (d, J = 8.4 Hz, 2H), 7.41 (d, J = 8.4 Hz, 2H), 7.53 (s, 2H), 7.67 (t, J = 13.2 Hz, 1H), 7.95 (t, J = 12.8 Hz, 2H). 13 C NMR (100MHz, DMSO-d6): δ29.93,32.60,49.09,104.48,111.47,119.96122.66,128.61,128.71,130.33,130.35,130.37,141.50,142.42.

[0078] The preparation process of compounds T800-FI, T800-FF, T800-II and T800-BrBr refers to T800-FBr, and the product structures have been identified.

[0079] Example 2-Example 7 tested the application performance of T800-FBr, T800-FI, T800-FF, T800-II and T800-BrBr. For the convenience of description, T800-X1X2 is used as a general term for T800-FBr, T800-FI, T800-II and T800-BrBr to describe the preparation and testing process of the samples.

[0080] Example 2: Singlet oxygen yield test of T800-FBr, T800-FI, T800-FF, T800-BrBr and T800-II.

[0081] Preparation of the test system: prepare 1,3-diphenylisobenzofuran (DPBF) mother solution with dichloromethane solution; dissolve each T800-X1X2 in dimethyl sulfoxide to prepare a T800-X1X2 mother solution with a concentration of 10 mM; dissolve ICG in dimethyl sulfoxide to obtain an ICG mother solution with a concentration of 10 mM.

[0082] In the singlet oxygen yield test involving T800-X1X2, 3 mL of dichloromethane was taken into a quartz dish, and a certain amount of DPBF mother liquor was added thereto until the ultraviolet absorption intensity of DPBF at 415 nm was about 1.0. Then a certain amount of T800-X1X2 mother liquor was added into the quartz dish until the ultraviolet absorption intensity of T800-X1X2 at 760 nm was about 0.3. Subsequently, the quartz dish containing the above solution was placed under an 808nm laser for intermittent irradiation, the illumination power was 3mW, the interval illumination time was 30s, and the total time was 300s. Then, the decrease in the ultraviolet absorption intensity of DPBF at 415nm after each interval illumination was measured; then, 3mL of dichloromethane was taken again in the quartz dish, and a certain amount of DPBF mother liquor was added until its ultraviolet absorption intensity at 415nm was about 1.0, and then a certain amount of ICG mother liquor was added to the quartz dish until the ultraviolet absorption intensity of ICG at 808nm was about 0.3. Then, the quartz dish containing the above solution was placed under an 808nm laser for intermittent irradiation, the irradiation power was 3mW, the interval irradiation time was 30s, and the total time was 300s. Then, the decrease in the ultraviolet absorption intensity of DPBF at 415nm after each interval irradiation was measured. Finally, the singlet oxygen yield of T800-X1X2 was calculated based on the measured decrease in the ultraviolet absorption intensity of DPBF, and was obtained by Figure 1It can be seen that the singlet oxygen yield of T800-II is the highest at 12.231%, and the singlet oxygen yield of T800-FI is 7.238%, which proves that the introduction of heavy atoms can increase the singlet oxygen yield, and a certain level of singlet oxygen production can still be maintained when a single heavy atom exists.

[0083] Example 3: Confocal fluorescence imaging test of T800-FBr, T800-FI, T800-FF, T800-BrBr and T800-II cell uptake

[0084] Preparation of the test system: Dissolve each T800-X1X2 in dimethyl sulfoxide solution to prepare a 10mM T800-X1X2 mother solution. Then take 2μL of the mother solution, dissolve it in 2mL of culture medium and shake it evenly to prepare a 10μM T800-X1X2 incubation solution for use. At this time, the pH value of the incubation solution is 7.4-7.5, and the temperature is 37°C.

[0085] In the process of detecting the in vitro cell uptake efficiency of T800-X1X2, the culture medium containing T800-X1X2 was added to the confocal dish containing MCF-7 living cells with a growth density of about 60%, and the change of the fluorescence intensity of the cells incubated with T800-X1X2 was measured over time. Figure 2 It can be seen that the cell fluorescence reaches its peak at about 1 hour, and T800-FI has the strongest fluorescence intensity among the five molecules, indicating that the target molecule T800-FI has the highest cellular uptake efficiency and is easier to achieve fluorescence-guided photodynamic therapy.

[0086] Example 4: Cytotoxicity test of T800-FBr, T800-FI, T800-BrBr and T800-II

[0087] Test system preparation: Dissolve each T800-X1X2 in dimethyl sulfoxide solution to prepare a 10mM T800-X1X2 mother solution, then take 10μL of the 10mM mother solution and dissolve it in 90μL of dimethyl sulfoxide solution to prepare a 1mM mother solution for use. Take 0μL, 0.5μL, 1μL, 1.5μL, 2μL, 3μL, 4μL, 5μL of the 1mM mother solution and dissolve them in 1mL of culture medium to prepare 0μM, 0.5μM, 1μM, 1.5μM, 2μM, 3μM, 4μM, 5μM incubation solutions for use. At this time, the pH value of the incubation solution is 7.4-7.5 and the temperature is 37°C.

[0088] A certain density of 4T1 cells, HepG-2 cells and MCF-7 cells were planted in a 96-well plate. When the cell growth density reached more than 80%, 100 μL of the above-configured T800-X1X2 culture medium of different concentrations was added to each well and incubated for 1 hour. Subsequently, continuous illumination was performed at 808 nm and 100 mW for 5 minutes, and the cells were transferred to a 37°C constant temperature incubator and incubated for 24 hours. The residual culture medium was removed, and then 100 μL of 5 mg / mL thiazolyl blue culture medium was added to each well, and the cells were incubated at 37°C for 4 hours. The residual culture medium was removed, and 100 μL of dimethyl sulfoxide was added to each well. The absorbance at 490 nm was immediately tested, and the survival rate of cells incubated with different photosensitizers was calculated. Figure 3 It can be seen that although the phototoxicity of the target molecule is lower than that of the comparison molecule, the target molecule T800-FI can still achieve a phototoxicity similar to that of the comparison molecule T800-II due to its higher cellular uptake efficiency.

[0089] Example 5: Time-dependent fluorescence imaging test of T800-FBr and T800-FI in the parathyroid gland

[0090] Preparation of test system: T800-FBr and T800-FI were dissolved in dimethyl sulfoxide to obtain 100mM T800-FBr mother solution and T800-FI mother solution, respectively. Then, 10μL of the 100mM mother solution was taken and dissolved in 990μL PBS solution to prepare a 1mM mother solution for use.

[0091] Experimental animal preparation: The experimental animals were approved by the Animal Experiment Ethics Committee of Dalian University of Technology (DUT20210902). The mouse imaging and treatment experimental operations were carried out in accordance with the "Guidelines for the Care and Use of Laboratory Animals of Dalian Medical University". 4-5 week old BALB / c female mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd.

[0092] In the time-dependent fluorescence imaging test of parathyroid tissue by T800-FBr and T800-FI, 100 μL of 1 mM T800-FBr PBS stock solution and T800-FI PBS stock solution were injected into mice through the tail vein to measure the changes in the fluorescence intensity of the parathyroid glands over time. Figure 4 As shown, the fluorescence intensity and retention time of T800-FI in the target molecule are longer, which is consistent with the previous cell uptake experiment and indicates that the target molecule reaches the parathyroid gland.

[0093] Example 6: Test of blood calcium concentration of rat parathyroid gland after phototherapy with T800-FBr and T800-FI

[0094] Preparation of test system: dissolve T800-FBr and T800-FI in dimethyl sulfoxide solution respectively to prepare 100mM T800-FBr mother solution and T800-FI mother solution, take 10μL of 100mM T800-X1X2 mother solution respectively, dissolve it in 990μL PBS solution to prepare 1mM T800-X1X2 mother solution; take 50μL of 100mM T800-X1X2 mother solution, dissolve it in 950μL PBS solution to prepare 5mM T800-X1X2 mother solution for use.

[0095] Experimental animal preparation: The experimental animals were approved by the Animal Experiment Ethics Committee of Dalian University of Technology (DUT20210902). The experimental procedures of rat treatment experiments were carried out in accordance with the Guidelines for the Care and Use of Laboratory Animals of Dalian Medical University. 250-300 g SD male rats were purchased from Liaoning Changsheng Biotechnology Co., Ltd.

[0096] In the test of blood calcium concentration after T800-FBr and T800-FI phototherapy of parathyroid tissue in rats, 200 μL of prepared T800-X1X2 (T800-FBr, T800-FI) stock solution with concentrations of 1 mM and 5 mM was injected into rats (about 300 g) through the tail vein. After incubation for 12 hours, an 808 nm laser was used to continuously illuminate the rats at a light power of 200 mW for 10 minutes. Blood was collected from each group at 0 h, 6 h, 24 h, 36 h, 3 d, 5 d, and 10 d to measure the blood calcium concentration of the rats. Figure 5 It can be seen that after photodynamic therapy, the blood calcium concentration of rats in each group decreased, indicating that the parathyroid glands of rats were ablated under photodynamic therapy, and the decrease in blood calcium concentration of rats in the T800-FI group was greater than that in the T800-FBr group, proving that T800-FI has the best photodynamic ablation effect on the parathyroid glands.

[0097] Example 7: Test of PTH concentration in rat parathyroid gland after phototherapy with T800-FBr and T800-FI The test system, phototherapy method, and blood sampling scheme were the same as those in Example 6. After blood was collected from each group, the PTH concentration of the rats was measured. Figure 6 It can be seen that after photodynamic therapy, the PTH concentration of rats in each group decreased, indicating that the parathyroid glands of rats were ablated under photodynamic therapy, and the decrease in PTH concentration of rats in the T800-FI group was greater than that in the T800-FBr group, proving that T800-FI has the best photodynamic ablation effect on the parathyroid glands.

Claims

1. Compounds of formula I: in, X1 and X2 are each independently F, Br or I, and X1 and X2 are not F or Br at the same time; Y - is an anion, and the anion has the same charge number as the cation in formula I; Y - F - , Cl - Br - ,I - 、BF4 - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - or CF3SO3 - .

2. The compound according to claim 1, characterized in that Select from the following structures:

3. The compound according to claim 1, characterized in that Select from the following structures:

4. The compound according to claim 1, characterized in that The compound of formula I is selected from the following structures:

5. The compound according to any one of claims 1 to 4, characterized in that Y - F - , Cl - , Br or I - .

6. The compound according to any one of claims 1 to 4, characterized in that Y - is I - .

7. Use of the compound according to any one of claims 1 to 4 in the preparation of photodynamic drugs.

8. Use of the compound according to any one of claims 1 to 4 in the preparation of a drug for treating cancer or parathyroid disease.

9. A method for preparing a compound of formula I, characterized in that: Using heptamethine cyanine dye as the photosensitizer matrix, the synthesis reaction is as follows: The synthesis steps are as follows: Compound A and pentadienaldehyde diphenylamine hydrochloride react in acetic anhydride to obtain compound B; In an inert gas atmosphere, compound B and compound C are reacted in acetic anhydride in the presence of a catalyst at room temperature to obtain a compound of formula I; The catalyst is one of sodium acetate and pyridine; Y - is an anion, and the anion has the same charge number as the cation in formula I; Y - F - , Cl - Br - ,I - 、BF4 - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - or CF3SO3 - .

10. The preparation method according to claim 9, characterized in that: Y - F - , Cl - , Br or I - .

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