CN111808111A

CN111808111A - Iridium complex, preparation method thereof and photodynamic therapeutic drug

Info

Publication number: CN111808111A
Application number: CN202010546775.4A
Authority: CN
Inventors: 张平玉; 陈海杰; 任奇志; 张黔玲
Original assignee: Shenzhen University
Current assignee: Shenzhen University
Priority date: 2020-06-16
Filing date: 2020-06-16
Publication date: 2020-10-23

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to an iridium complex, wherein the structural general formula of the iridium complex is shown as the following formula I or formula II:

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈Each independently selected from: one of hydrogen, alkoxy and aryl; y is₁、Y₂Is an ion which is counter-balanced with the iridium complex host; n1, n2, m1 and m2 are ionic valences, and the absolute value of n1 is equal to the absolute value of n2 and the absolute value of m1Absolute value of m 2. The iridium complex has good water solubility, can be specifically combined with cell lysosomes, can generate singlet oxygen under the illumination condition, has obvious photodynamic treatment effect on cells, does not generate singlet oxygen under the dark condition, has no toxicity, and has wide application prospect in photodynamic treatment medicines.

Description

Iridium complex, preparation method thereof and photodynamic therapeutic drug

Technical Field

The application belongs to the technical field of biological medicines, and particularly relates to an iridium complex and a preparation method thereof, and a photodynamic therapy medicament.

Background

Photodynamic therapy (PDT) is a relatively novel noninvasive tumor treatment method, and relies on a light source with a specific wavelength to irradiate and activate a photosensitizer in tumor tissues to generate Reactive Oxygen Species (ROS) such as singlet oxygen with biological toxicity, so as to oxidize and damage tumors, virus-infected cells and other hyperproliferation cells, activate antitumor and anti-virus immunity, damage blood vessels, kill bacteria, fungi and viruses and eliminate inflammation. The medical scientist finds that the acid extract of the blood of the mammal can effectively inhibit the growth of the tumor after being irradiated by light, and the first generation photosensitizer hematoporphyrin is generated after the process is improved. However, the clinical efficacy of hematoporphyrins is unstable due to unclear composition and activity ability in photodynamic therapy and differences in preparation processes. The second generation photosensitizer is also porphyrin derivative, and has specific chemical structure, stable and repeatable clinical curative effect compared with the first generation photosensitizer. However, improvements in light absorption in the water-soluble biological environment spectrum, selectivity for cancer cells, photostability, avoidance of photodegradation to bilirubin, rate of in vivo metabolism, localization, etc., of such photosensitizers are still needed.

In recent years, heavy metals have been found to have a significant effect on the efficiency of PDT. Pd^II(WST11)，Lu^III(Lutex) and Sn^IVPhotosensitizers in which metals such as (Purlytin) are coordinated to porphyrins have been developed and tested clinically. With the increasingly stringent requirements for photosensitizers in medical research, Ru^II、Os^IIOr Re^IOf poly-pyridyl low spin of⁶The use of configured metal complexes as photosensitizers for PDT has also attracted attention. However, the existing photosensitizer of the metal complex has poor water solubility, so that the problems of low cellular uptake efficiency, long cellular penetration time, large dosage in clinical treatment experiments and the like are caused.

Disclosure of Invention

The application aims to provide an iridium complex, a preparation method thereof and a photodynamic therapy medicament, and aims to solve the problems of poor water solubility, low cell uptake rate and long cell permeation time of the existing metal complex photosensitizer to a certain extent.

In order to achieve the purpose of the application, the technical scheme adopted by the application is as follows:

in a first aspect, the present application provides an iridium complex, wherein the structural general formula of the iridium complex is shown as formula I or formula II below:

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈Each independently selected from: one of hydrogen, alkoxy and aryl; y is₁、Y₂Is an ion which is counter-balanced with the iridium complex host; n1, n2, m1 and m2 are ionic valences, and the absolute value of n1 is equal to the absolute value of n2, and the absolute value of m1 is equal to the absolute value of m 2.

In a second aspect, the present application provides a method for preparing an iridium complex, comprising the steps of:

mixing the compound A with concentrated sulfuric acid, performing sulfonation reaction, and separating to obtain a sulfonated compound B;

neutralizing the compound B by using a first alkaline salt, and separating to obtain a compound C;

dissolving the compound C and an iridium precursor in a first organic solvent, carrying out a coordination reaction, and separating to obtain a first iridium complex;

wherein, the structural general formula of the compound A is as follows:

or

When the structural formula of the compound A is A1, the structural formula of the first iridium complex is I-1; when the structural formula of the compound A is A2, the structural formula of the first iridium complex is II-1:

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈Each independently selected from: hydrogen or alkoxy, Y₁、Y₂Is an ion which is counter-balanced with the iridium complex host; n1, n2, m1 and m2 are ionic valences, and the absolute value of n1 is equal to the absolute value of n2, and the absolute value of m1 is equal to the absolute value of m 2.

In a third aspect, the present application provides a method for preparing an iridium complex, comprising the steps of:

performing chloromethylation reaction on the compound D by adopting polyformaldehyde and hydrogen chloride, and separating to obtain a compound E;

mixing the compound E with concentrated sulfuric acid, performing sulfonation reaction, and separating to obtain a sulfonated compound F;

neutralizing the compound F by using a second alkaline salt, and separating to obtain a compound G;

dissolving a compound G and triphenylphosphine in a second organic solvent, carrying out thermal reaction, and separating to obtain a compound H;

dissolving the compound H and an iridium precursor in a third organic solvent, carrying out a coordination reaction, and separating to obtain a second iridium complex I grafted with triphenylphosphine methyl;

wherein the structural formula of the compound D is as follows:

or

When the structural formula of the compound D is D1, the structural formula of the second iridium complex is I-2; when the structural formula of the compound D is D2, the structural formula of the second iridium complex is II-2:

wherein, Y₁ ^-、Y₂ ^-Is an ion which is counter-balanced with the iridium complex host.

In a fourth aspect, the present application provides a photodynamic therapy medicament comprising the above iridium complex, or an iridium complex prepared by the above method.

The iridium complex provided by the first aspect of the application can generate singlet oxygen under the illumination condition of 400 nm-600 nm, and has an obvious photodynamic treatment effect on cells, namely, obvious phototoxicity. And under the dark condition, no singlet oxygen is generated, no toxicity exists, namely no dark toxicity exists, and the application prospect in photodynamic therapy medicines is wide.

According to the preparation method of the iridium complex provided by the second aspect of the application, the water solubility of the iridium complex can be remarkably increased by introducing the strong-polarity sulfonic group to the porphyrin or phthalocyanic ring. In addition, through coordination with metal iridium, the iridium complex is beneficial to generating molecular oxygen under the illumination condition, the photodynamic effect of the complex is enhanced, the three-dimensional structure of the complex is changed, and two molecules of water are introduced for coordination, so that the water solubility of the complex is further enhanced, the uptake rate of the complex by cells is improved, and the permeation time of the complex to the cells is shortened.

According to the preparation method of the iridium complex provided by the third aspect of the application, the water solubility of the iridium complex can be remarkably increased by introducing the strong-polarity sulfonic group to the porphyrin or phthalocyanyl ring. And the triphenylphosphine substituent capable of targeting cell mitochondria is introduced, so that the cell targeting property of the complex is enhanced, the iridium complex is favorable for generating molecular oxygen under the illumination condition, the photodynamic effect of the complex is enhanced, the uptake rate of the complex by cells is improved, and the permeation time of the complex to the cells is shortened.

The photodynamic therapy medicament provided by the fourth aspect of the application contains the iridium complex which has obvious phototoxicity and no dark toxicity under the illumination condition, and the complex has good water solubility and can be specifically combined with lysosomes or mitochondria of cells. Therefore, the photodynamic therapy medicament provided by the application has the advantages of fast penetration in cells, good stability of combination with the cells, capability of quickly and accurately treating disease tissues through the toxicity of the iridium complex under illumination and good photodynamic effect.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 is a scheme showing the synthesis scheme of methoxy-porphyrin-iridium complex in example 1 of the present application;

FIG. 2 is a scheme showing the synthesis scheme of triphenylphosphine-porphyrin-iridium complex in example 2 of the present application;

FIG. 3 is a scheme showing the synthesis scheme of methoxy-phthalocyanyl-iridium complex in example 3 of the present application;

FIG. 4 is a graph of the UV and fluorescence spectra of methoxy-porphyrin-iridium complex of example 1 of the present application;

FIG. 5 is an infrared spectrum of methoxy-porphyrin-iridium complex of example 1 of the present application.

FIG. 6 is a confocal microscopic image of methoxy-porphyrin-iridium complex in hepatoma cells Hep-G2 in example 1 of the present application.

FIG. 7 is a graph of co-staining factor of methoxy-porphyrin-iridium complex with the lysosomal green dye LTG in hepatoma cells Hep-G2 in example 1 of the present application.

FIG. 8 is a graph showing co-staining coefficients of methoxy-porphyrin-iridium complex and a mitochondrial green dye MTG in hepatoma cells Hep-G2 in example 1 of the present application.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects to be solved by the present application more clearly apparent, the present application is further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present application and are not intended to limit the present application.

In this application, the term "and/or" describes an association relationship of associated objects, meaning that there may be three relationships, e.g., a and/or B, which may mean: a is present alone, A and B are present simultaneously, and B is present alone. Wherein A and B can be singular or plural. The character "/" generally indicates that the former and latter associated objects are in an "or" relationship.

In the present application, "at least one" means one or more, "a plurality" means two or more. "at least one of the following" or similar expressions refer to any combination of these items, including any combination of the singular or plural items. For example, "at least one (a), b, or c", or "at least one (a), b, and c", may each represent: a, b, c, a-b (i.e., a and b), a-c, b-c, or a-b-c, wherein a, b, and c may be single or plural, respectively.

It should be understood that, in various embodiments of the present application, the sequence numbers of the above-mentioned processes do not mean the execution sequence, some or all of the steps may be executed in parallel or executed sequentially, and the execution sequence of each process should be determined by its function and inherent logic, and should not constitute any limitation to the implementation process of the embodiments of the present application.

The terminology used in the embodiments of the present application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used in the examples of this application and the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

The weight of the related components mentioned in the description of the embodiments of the present application may not only refer to the specific content of each component, but also represent the proportional relationship of the weight among the components, and therefore, the content of the related components is scaled up or down within the scope disclosed in the description of the embodiments of the present application as long as it is scaled up or down according to the description of the embodiments of the present application. Specifically, the mass in the description of the embodiments of the present application may be in units of mass known in the chemical industry, such as μ g, mg, g, and kg.

The terms "first" and "second" are used for descriptive purposes only and are used for distinguishing purposes such as substances from one another, and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. For example, a first XX may also be referred to as a second XX, and similarly, a second XX may also be referred to as a first XX, without departing from the scope of embodiments of the present application. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature.

In a first aspect of the embodiments of the present application, an iridium complex is provided, where a structural general formula of the iridium complex is shown as formula I or formula II below:

According to the iridium complex provided by the first aspect of the application, porphyrin or phthalocyane with four sulfonic acid groups is used as a main ligand, and the iridium complex is formed by coordination with metal iridium. On one hand, the coordinated iridium metal in the iridium complex deviates from the plane of porphyrin or phthalonitrile molecules, so that the molecules are converted from a two-dimensional plane structure to a three-dimensional structure, and the three-dimensional structure of the complex is changed. Therefore, the steric hindrance of molecules is increased, the acting force between molecules is weakened, the loss of excited state capacity caused by the formation of dimers between iridium complexes is avoided, the triplet state quantum yield and the service life of the iridium complexes are further ensured, the iridium complexes are more favorable for generating molecular oxygen under the illumination condition, and the photodynamic effect of the complexes is enhanced. On the other hand, the iridium metal center in the iridium complex is coordinated with two water molecules besides being coordinated with the N atom on the porphyrin or phthalo cyanide ring, which is beneficial to increasing the water solubility of the iridium complex. On the other hand, four sulfonic acid groups connected to porphyrin or phthalonitrile have strong polarity, so that the water solubility of the iridium complex is obviously improved; meanwhile, substituents such as alkoxy, aryl and the like connected on porphyrin or phthalonitrile can increase the water solubility of the iridium complex, and can be selectively connected with a group with targeting property, thereby being beneficial to improving the specificity of the iridium complex. The iridium complex provided by the embodiment of the application can generate singlet oxygen under the illumination condition of 400-600 nm, and has an obvious photodynamic treatment effect on cells, namely obvious phototoxicity. And under the dark condition, no singlet oxygen is generated, no toxicity exists, namely no dark toxicity exists, and the application prospect in photodynamic therapy medicines is wide.

In some embodiments, Y₁、Y₂The valence of the complex host ion is conserved with the overall valence of the counter ion, namely n1 is equal to n2 and m1 is equal to m 2. The counter ion is an anion when the iridium complex body is positively charged and a cation when the iridium complex body is negatively charged.

In some embodiments, Y⁺Selected from: sodium ion, potassium ion, ammonium ion, and chloride ion. In some embodiments, when the iridium complex host ion is positively charged, the counter ion is an anion such as chloride; when the iridium complex main body ions are negatively charged, the counter ions adopt cations such as sodium ions, potassium ions, ammonium ions and the like. In the embodiment of the application, counter ions such as sodium ions, potassium ions, ammonium ions and chloride ions are combined with the iridium complex main body, and the counter ions are ions with abundant contents in organisms, have good fusion effect with organism cells and have no biological toxicity. Moreover, the counter ions can not react with alkalescent body fluid after being ionized in the body fluid, and hydrogen is avoidedWhen the ion is used as a counter ion, it is ionized to be acidic in a body fluid, and reacts with a basic body fluid in a living body, thereby exerting an adverse effect on the living body.

In some embodiments, R in the iridium complex₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈Each independently selected from: methoxy or triphenylphosphine methyl. The methoxyl group has good binding specificity with lysosomes in cells, so that the complex has lysosome targeting property, can target the lysosomes of the cells, and has good binding stability and strong specificity. The triphenylphosphine substituent can target mitochondria of cells, and the mitochondria are the places with respiration and adenosine triphosphate ATP synthesis, are the most sufficient place of molecular oxygen in the cells, are beneficial to the photosensitizer to generate more molecular oxygen under the illumination condition, and further improve the cytotoxicity of the iridium complex.

In some embodiments, R₁、R₂、R₃、R₄Simultaneously selected from: methoxy or triphenylphosphine methyl; r₅、R₆、R₇、R₈Simultaneously selected from: methoxy or triphenylphosphine methyl, and the same targeting substituent is connected on the porphyrin ring or phthalocyanine ring of the iridium complex, so that the targeting specificity and stability of the iridium complex in organisms can be enhanced.

In some embodiments, the iridium complex is a porphyrin-iridium complex, including the following structural formula:

R₁、R₂、R₃、R₄simultaneously selected from: methoxy or triphenylphosphine methyl. When R is₁、R₂、R₃、R₄When simultaneously selected from methoxy, Y₁Selected from: sodium ions, potassium ions or ammonium ions. When R is₁、R₂、R₃、R₄When simultaneously selected from triphenylphosphine methyl, Y₁Selected from chloride ions.

In some embodiments, the iridium complex is a phthalocyaniridium complex comprising the formula:

R₅、R₆、R₇、R₈simultaneously selected from: methoxy or triphenylphosphine methyl. When R is₅、R₆、R₇、R₈When simultaneously selected from methoxy, Y₂Selected from: sodium ions, potassium ions or ammonium ions. When R is₅、R₆、R₇、R₈When simultaneously selected from triphenylphosphine methyl, Y₂Selected from chloride ions.

The iridium complex provided by the embodiment of the application can be prepared by the following method.

A second aspect of the embodiments of the present application provides a method for preparing an iridium complex, including the following steps:

s10, mixing the compound A with concentrated sulfuric acid, performing sulfonation reaction, and separating to obtain a sulfonated compound B;

s20, neutralizing the compound B by using a first alkaline salt, and separating to obtain a compound C;

s30, dissolving the compound C and an iridium precursor in a first organic solvent, carrying out a coordination reaction, and separating to obtain a first iridium complex;

wherein, the structural general formula of the compound A is as follows:

or

wherein the content of the first and second substances,R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈each independently selected from: hydrogen or alkoxy, Y₁、Y₂Is an ion which is counter-balanced with the iridium complex host; n1, n2, m1 and m2 are ionic valences, and the absolute value of n1 is equal to the absolute value of n2, and the absolute value of m1 is equal to the absolute value of m 2.

According to the preparation method of the iridium complex provided by the second aspect of the application, the compound A is used as a raw material, and is subjected to sulfonation reaction with concentrated sulfuric acid, and a sulfonic group is connected to porphyrin or phthalocyanic acid, so that a sulfonated compound B is obtained. And then neutralizing the sulfonated compound B by using a first alkaline salt, and introducing counter cations into the complex to enable the complex to be electrically neutral to obtain a compound C. And then carrying out coordination reaction on the compound C and an iridium precursor to coordinate iridium metal with a nitrogen atom in porphyrin or phthalo cyanide ring, and simultaneously introducing two coordinated water molecules into the iridium metal to obtain a first iridium complex. According to the preparation method of the iridium complex, the water solubility of the iridium complex can be remarkably improved by introducing the strong-polarity sulfonic group to the porphyrin or phthalocyanyl ring. In addition, through coordination with metal iridium, the iridium complex is beneficial to generating molecular oxygen under the illumination condition, the photodynamic effect of the complex is enhanced, the three-dimensional structure of the complex is changed, and two molecules of water are introduced for coordination, so that the water solubility of the complex is further enhanced, the uptake rate of the complex by cells is improved, and the permeation time of the complex to the cells is shortened.

Specifically, in step S10, compound a and concentrated sulfuric acid are mixed and then subjected to sulfonation reaction, and four sulfonic acid groups are introduced to the branched chain of the porphyrin ring, where the sulfonic acid groups have strong polarity, so as to increase the water solubility of the complex and improve the uptake efficiency of the complex by cells. In addition, the compound A of the embodiment of the application is connected with hydrogen and alkoxy substituent groups, wherein the alkoxy groups such as methoxy group and the like have stronger binding specificity with lysosomes in cells, so that the complex can target the lysosomes of the cells, and has lysosome targeting, good binding stability and strong specificity.

In some embodiments, the mass to volume ratio of compound a to concentrated sulfuric acid is 1 g: (20-30) ml, and the raw material components in the proportion can fully ensure the sulfonation effect of concentrated sulfuric acid on the compound A.

In some embodiments, the sulfonation reaction between compound a and concentrated sulfuric acid is an electrophilic substitution reaction, and the attack of the sulfonic acid group on the large pi bond on the relatively stable benzene ring in compound a needs to be performed at a relatively high temperature of 100 ℃. Meanwhile, concentrated sulfuric acid adopted in the sulfonation reaction has strong zinc oxide, and if the temperature is higher than 110 ℃, the compound A is partially carbonized due to overhigh temperature and is mixed and adhered with the product, so that the yield of the complex is reduced, and the difficulty in separating and purifying the product is increased. In addition, the sulfonation reaction time is 5-8 hours, and the sufficient sulfonation of the compound A by concentrated sulfuric acid is fully ensured.

When the structural formula of the compound A is A1, the structural formula of the sulfonated compound B is B1; when the structural formula of the compound A is A2, the structural formula of the sulfonated compound B is B2, and the reaction formula is as follows:

in some embodiments, the step of mixing compound a with concentrated sulfuric acid followed by sulfonation comprises:

s11, according to the volume ratio of the mass of the compound A to concentrated sulfuric acid of 1 g: (8-15) ml, mixing and grinding the compound A and concentrated sulfuric acid to obtain a pasty mixture;

s12, adding the residual concentrated sulfuric acid into the pasty mixture, and reacting for 5-8 hours at the temperature of 100-110 ℃.

The mass ratio and the volume ratio of the embodiment of the application are 1 g: (8-15) ml of the compound A and concentrated sulfuric acid are mixed and ground to form a pasty mixture, so that the raw material components are mixed more uniformly, and the sulfonation reaction is facilitated. If the concentrated sulfuric acid content is too high or too low, the grinding process is not favorable. And then mixing the residual concentrated sulfuric acid with the pasty mixture, and reacting for 5-8 hours at the temperature of 100-110 ℃ to enable the concentrated sulfuric acid to fully sulfonate the compound A to obtain a sulfonated compound B.

In some embodiments, the sulfonated product is sequentially diluted, filtered, extracted and dried to obtain the sulfonated compound B. In some specific embodiments, the sulfonated reaction system is kept stand for 24-48 hours, deionized water is slowly added for dilution after the substance components are stable, and then the dilution is also filtered to remove the unreacted compound A; and adding acetone into the filtrate for extraction for 2-3 times, and removing the acetone to obtain a dried sulfonated compound B.

Specifically, in step S20, since the hydrogen ions are ionized in the body fluid to become acidic and react with the body fluid with a more basic organism, the counter ions of the complex are changed into metal ions and the like with abundant contents in the organism instead of the hydrogen ions by the neutralization treatment of the sulfonated compound B with the basic salt in the embodiment of the present application, so that the biocompatibility of the complex is improved and the biotoxicity is reduced.

When the structural formula of the compound A is A1, the structural formula of the compound C is C1; when the structural formula of the compound A is A2, the structural formula of the compound C is C2, and the reaction formula is as follows:

wherein R is₁、R₂、R₃、R₄Each independently selected from: one of hydrogen and alkoxy, and Y is a counter ion introduced by the neutralization reaction of a basic substance.

In some embodiments, the basic salt is selected from: NH (NH)₄HCO₃、K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃The basic salts can neutralize the acidity of the sulfonated compound B, so that counter ions in the complex are converted from hydrogen ions into cations rich in living bodies such as sodium ions, potassium ions, ammonium ions and the like, and the biocompatibility of the complex is improved.

In some embodiments, the step of neutralizing comprises: adjusting the pH value of the water solution of the sulfonated compound B to 6.0-7.0 by using alkaline salt, fully neutralizing hydrogen ions in the sulfonated compound B, and separating to obtain a compound C.

In some embodiments, the neutralized product is subjected to dialysis concentration treatment, recrystallization treatment and drying treatment in sequence, and compound C is isolated. In some embodiments, the sulfonated compound B is dissolved in water and then: NH (NH)₄HCO₃、K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃After the pH value of the aqueous solution is adjusted to 6-7, dialyzing the solution; and then recrystallizing for 2-3 times by using a dehydrated methanol/ethanol mixed solvent, filtering and then drying in vacuum to obtain a compound C.

Specifically, in step S30, the compound C and the iridium precursor are dissolved in a first organic solvent, and subjected to a coordination reaction, followed by separation to obtain a first iridium complex. According to the embodiment of the application, the compound C and an iridium precursor are dissolved in an organic solvent, and coordination reaction is carried out on iridium metal and nitrogen atoms on porphyrin or phthalonitrile rings in the complex, so that the iridium metal can be coordinated with two molecules of water at the same time, and a first iridium complex in which the iridium metal is coordinated with water and a porphyrin complex or phthalonitrile ligand is obtained.

In the embodiment of the application, when the structural formula of the compound A is A1, the first iridium complex is a porphyrin-iridium ligand, and the structural formula is I-1; when the structural formula of the compound A is A2, the first iridium complex is phthalo cyanide-iridium ligand with the structural formula of II-1, and the reaction formula is as follows:

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈Each independently selected from: hydrogen or alkoxy, Y₁、Y₂Is an ion which is counter-balanced with the iridium complex host; n1, n2, m1 and m2 are ionic valences, and n1 is n2, and m1 is m 2.

In some embodiments, the iridium precursor is selected from: at least one of 1, 5-cyclooctadiene iridium chloride (I) dimer, chlorobis (cyclooctene) iridium (I) dimer and carbonyl bis (triphenylphosphine) iridium chloride (I), wherein iridium metal in the iridium precursor can be coordinated with nitrogen atoms in porphyrin or phthalocyanin ring of compound C to form porphyrin-iridium complex or phthalocyanin-iridium complex, and can be coordinated with water molecules to further improve the water solubility of the complex.

In some embodiments, the compound C and the iridium precursor are dissolved in at least one organic solvent selected from ethylene glycol, glycerol and dimethyl sulfoxide, and the organic solvents have good solubility for both the compound C and the iridium precursor, so that a stable solvent system is provided for coordination between the compound C and the iridium precursor. In some specific embodiments, the compound C and the iridium precursor are dissolved in ethylene glycol for coordination reaction, and the ethylene glycol has good solubility to the compound C and the iridium precursor, and low viscosity, which is beneficial to the full mixing reaction of the compound C and the iridium precursor; the boiling point is low and easy to remove; and the coordination with iridium metal can not occur, thereby being beneficial to the separation and purification of the target product.

In some embodiments, the molar ratio of compound C to iridium precursor is 1: (1-1.2) dissolving the compound C and an iridium precursor in an organic solvent for coordination reaction, wherein the proportion ensures sufficient coordination of the compound C and the iridium precursor, and simultaneously avoids the phenomenon that a large amount of unreacted raw material substances or other byproducts are left in a reaction system due to excessive addition of raw materials, and the separation difficulty of a target product is increased.

In some embodiments, the step of performing a coordination reaction comprises: and dissolving the compound C and an iridium precursor in an organic solvent, and carrying out a coordination reaction at the temperature of 120-160 ℃ for 5-6 hours. If the temperature is too high, insoluble coking substances are easy to appear on the wall of the container of the reaction system, and the yield of the target product is reduced; if the temperature is too low, the oxidation effect of monovalent iridium is reduced, and it takes a longer time to follow the completion of the reaction, resulting in low reaction efficiency.

In some embodiments, the coordination reaction may also be performed in an inert atmosphere such as nitrogen, argon, etc., which is beneficial to eliminating interference of side reactions and improving the coordination reaction effect. Specifically, the molar ratio of the compound C to the iridium precursor is 1: (1-1.2) dissolving the compound C and an iridium precursor in organic solvents such as ethylene glycol, glycerol, dimethyl sulfoxide and the like; then reacting under the condition of inert atmosphere with the temperature of 120-160 ℃, tracking the reaction progress through ultraviolet, and confirming the reaction is complete when two groups of UV curves of the solution in the reaction system which are separated by 0.5h have no obvious difference.

In some embodiments, the step of isolating the iridium complex comprises: and sequentially carrying out drying treatment, washing treatment and concentration treatment on the product after the coordination reaction. In some embodiments, the step of isolating the iridium complex comprises: and removing the organic solvent in the product after the coordination reaction, washing the product for 2-3 times by using deionized water, collecting filtrate, and heating and concentrating the filtrate. Then, unreacted iridium precursor is removed by dialysis purification. Then obtaining the iridium complex by reduced pressure distillation.

A third aspect of the embodiments of the present application provides a method for preparing an iridium complex, including the following steps:

s40, performing chloromethylation reaction on the compound D by adopting polyformaldehyde and hydrogen chloride, and separating to obtain a compound E;

s50, mixing the compound E with concentrated sulfuric acid, performing sulfonation reaction, and separating to obtain a sulfonated compound F;

s60, neutralizing the compound F by using a second alkaline salt, and separating to obtain a compound G;

s70, dissolving the compound G and triphenylphosphine in a second organic solvent, carrying out thermal reaction, and separating to obtain a compound H;

s80, dissolving the compound H and an iridium precursor in a third organic solvent, performing a coordination reaction, and separating to obtain a second iridium complex I grafted with a triphenylphosphine methyl group;

wherein the structural formula of the compound D is as follows:

or

According to the preparation method of the iridium complex provided by the embodiment of the third aspect of the application, the compound D is used as a raw material, and chloromethyl is grafted on a benzene ring of the compound D through chloromethylation reaction, so as to provide conditions for subsequent grafting of triphenylphosphine substituent. Then, the sulfonated compound E is obtained by carrying out sulfonation reaction with concentrated sulfuric acid and connecting sulfonic groups on porphyrin or phthalocyane. And neutralizing the sulfonated compound E by using a second alkaline salt to enable the complex to be electrically neutral, thereby being beneficial to the connection of subsequent triphenylphosphine substituent groups. Compound G is thermally reacted with triphenylphosphine to attach triphenylphosphine to the methyl group, yielding compound H. And then carrying out coordination reaction on the compound H and an iridium precursor to coordinate iridium metal with a nitrogen atom in porphyrin or phthalo cyanide ring, and simultaneously introducing two coordinated water molecules into the iridium metal to obtain a second iridium complex I. According to the preparation method of the second iridium complex I, the water solubility of the iridium complex can be remarkably increased by introducing the strong-polarity sulfonic group to the porphyrin or phthalocyanyl ring. And the triphenylphosphine substituent capable of targeting cell mitochondria is introduced, so that the cell targeting property of the complex is enhanced, the iridium complex is favorable for generating molecular oxygen under the illumination condition, the photodynamic effect of the complex is enhanced, the uptake rate of the complex by cells is improved, and the permeation time of the complex to the cells is shortened.

Specifically, in step S40, a chloromethylation reaction is performed on the compound D using polyoxymethylene and hydrogen chloride to graft chloromethyl on the benzene ring of the compound D, so as to provide conditions for subsequent grafting of triphenylphosphine, and the compound E is obtained by separation.

In the examples, when the structural formula of the compound D is D1, the structural formula of the compound E is E1; when the structural formula of the compound D is D2, the structural formula of the compound E is E2, the reaction formula is as follows:

in some embodiments, the step of chloromethylating compound D comprises: dissolving polyformaldehyde, a compound D and hydrochloric acid in an organic solvent, reacting for 2-6 hours at the temperature of 60-100 ℃ under the condition of containing a chloride condensing agent, reacting a benzene ring in the compound D with polyformaldehyde and hydrogen chloride, replacing a hydrogen atom on aromatic hydrocarbon by chloromethyl, and separating to obtain a compound E. In some embodiments, the molar ratio of polyoxymethylene, compound D, and hydrochloric acid is 1: (4-5): (1-2), the chloromethylation of the compound D is fully ensured by the raw material components in the molar ratio.

In some embodiments, the polyoxymethylene is selected from: at least one of trioxymethylene, paraformaldehyde, formaldehyde dimethyl acetal, and chloromethyl ether. In some embodiments, the chloride salt condensing agent is selected from: at least one of zinc chloride, aluminum chloride and tin chloride. In the above embodiments of the present application, the polyoxymethylenes can be protonated and attack the benzene ring in the compound D under the action of hydrochloric acid; then grafting chloromethyl on benzene ring under the action of the chlorine salt condensing agent.

Specifically, in step S50, compound E is mixed with concentrated sulfuric acid and then subjected to sulfonation reaction, a strongly polar sulfonic acid group is grafted to the ortho-position of the chloromethyl group in compound E, the water solubility of the complex is improved, and sulfonated compound F is obtained by separation. According to the embodiment of the application, a sulfonic group is introduced into a compound before a triphenylphosphine substituent is grafted, and because subsequently grafted triphenylphosphine has charges, if triphenylphosphine is grafted to a sulfonic group, the introduced sulfonic group is preferentially connected to a benzene ring of triphenylphosphine, so that an expected structure cannot be obtained, and the targeting property of triphenylphosphine is damaged.

In some embodiments, the conditions of the sulfonation reaction include: reacting for 5-8 hours at the temperature of 100-110 ℃.

In some embodiments, the mass to volume ratio of compound E to concentrated sulfuric acid is 1 g: (20-30) ml.

In the above embodiments of the present application, the influence of the condition parameters of the sulfonation process is discussed in detail in the foregoing text and is not repeated again. When the structural formula of the compound D is D1, the structural formula of the compound F is F1; when the structural formula of the compound D is D2, the structural formula of the compound F is F2, and the reaction formula is as follows:

specifically, in step S60, compound F is neutralized with a second basic salt, and compound G is isolated.

In some embodiments, the step of neutralizing comprises: adjusting the pH of the water solution of the sulfonated compound F to 6.0-7.0 by using a second alkaline salt, and separating to obtain a compound G.

In some embodiments, the second basic salt is selected from: NH (NH)₄HCO₃、K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃At least one of (1).

In the above embodiments of the present application, the influence of the condition parameter of the neutralization process is discussed in detail in the foregoing text, and is not repeated again. When the structural formula of the compound D is D1, the structural formula of the compound G is G1; when compound D has the formula D2, compound G has the formula G2:

wherein Y is a counter ion introduced by neutralization.

Specifically, in step S70, compound G and triphenylphosphine are dissolved in a second organic solvent, and a thermal reaction is performed, so that triphenylphosphine is grafted on a methyl group instead of chlorine through the thermal reaction, and compound H is isolated.

In some embodiments, the conditions of the thermal reaction include: and dissolving the compound G and triphenylphosphine in a second organic solvent, heating and refluxing for 12-24 hours to ensure that the triphenylphosphine is fully grafted, and separating to obtain a compound H.

In some embodiments, the molar ratio of compound G to triphenylphosphine is 1: (4-5), and the molar ratio ensures sufficient reaction among substances.

In some embodiments, the second organic solvent is selected from: toluene and/or 1, 4-dioxane, and the solvents have good solvent property for the compound G and triphenylphosphine, and provide a solvent environment for the grafting reaction.

In the examples of the application, when the structural formula of the compound D is D1, the structural formula of the compound H is H1; when the structural formula of the compound D is D2, the structural formula of the compound H is H2, and the reaction formula is as follows:

wherein, PPh₃ ⁺Is composed of

Specifically, in step S80, compound H and an iridium precursor are dissolved in a third organic solvent, and a coordination reaction is performed, so that iridium metal is coordinated with a nitrogen atom on a porphyrin or phthalonitrile ring in the complex, and the iridium metal can be coordinated with two molecules of water at the same time, a crude product of the solvent is removed by distillation under reduced pressure, and the crude product is eluted and purified by a dichloromethane/methanol system, so as to obtain a second iridium complex I grafted with a triphenylphosphine methyl group.

In some embodiments, the step of coordinating comprises: and dissolving the compound H and the iridium precursor in a third organic solvent, and reacting for 12-24 hours at the temperature of 140-170 ℃. In some embodiments, the coordination reaction may also be performed in an inert atmosphere such as nitrogen, argon, etc., which is beneficial to eliminating interference of side reactions and improving the coordination reaction effect.

In some embodiments, the molar ratio of compound H to iridium precursor is 1: (1-1.2) ensuring sufficient coordination of iridium metal.

In some embodiments, the iridium precursor is selected from: at least one of 1, 5-cyclooctadiene iridium chloride (I) dimer, chlorobis (cyclooctene) iridium (I) dimer and carbonyl bis (triphenylphosphine) iridium chloride (I), wherein iridium metal in the iridium precursor can be coordinated with nitrogen atoms in porphyrin or phthalocyanine ring to form porphyrin-iridium complex or phthalocyanine-iridium complex, and can be coordinated with water molecules to further improve the water solubility of the complex.

In some embodiments, the third organic solvent is selected from: at least one of xylene, ethylene glycol and dimethyl sulfoxide.

In the above embodiment of the present application, when the structural formula of the compound D is D1, the second iridium complex I is a porphyrin-iridium ligand, and the structural formula is I-2; when the structural formula of the compound D is D2, the second iridium complex I is phthalo cyanide-iridium ligand with the structural formula II-2, and the reaction formula is as follows:

wherein, Y₁ ^-、Y₂ ^-Is an ion which is counter-balanced with the iridium complex host; the counterion may be a chloride.

In a fourth aspect, the embodiments of the present application provide a photodynamic therapy medicament, which contains the iridium complex described above, or the iridium complex prepared by the above method.

The photodynamic therapy medicament provided by the fourth aspect of the application contains the iridium complex which has obvious phototoxicity and no dark toxicity under the illumination condition; and the iridium complex has good water solubility and can be specifically combined with cell lysosomes, mitochondria and the like. Therefore, the photodynamic therapy medicament provided by the embodiment of the application has the advantages of fast penetration in cells, good stability of combination with the cells, capability of quickly and accurately treating disease tissues through the toxicity of the iridium complex under illumination and good photodynamic effect.

In order to make the details of the above-mentioned implementation and operation of the present invention clearly understood by those skilled in the art and to make the progress of the iridium complex and the preparation method thereof obvious, the technical solutions are illustrated by the following examples.

Example 1

A methoxy-porphyrin-iridium complex has a synthetic scheme shown in figure 1, and comprises the following preparation steps:

taken 4.00g of TMPP (5,10,15, 20-tetrakis- (4-methoxyphenyl) -porphyrin) and 40.0mL (98.0% H)₂SO₄) Ground into a uniform paste in a mortar. The paste was transferred to a 250mL round bottom flask, then 60mL 98% H was added₂SO₄The mixture was heated with stirring in an oil bath at 105 ℃ for 6 h.

② the mixture was placed in a 400mL beaker and allowed to stand at room temperature for 48 hours. Then 300mL of deionized water was added slowly and stirred. After cooling, the solution was filtered through a G5 sintered glass filter to remove unreacted TMPP, and 200mL of acetone was added to the filtrate and stirred to precipitate a precipitate. The resulting precipitate was washed 2 times with acetone. Acetone was removed and the solid product was dissolved in 150mL deionized water and saturated NaHCO₃Neutralized to purple. After the pH value is adjusted to 6.0, the solution is dialyzed and concentrated, and then is recrystallized twice by using a dehydrated methanol/ethanol mixed solvent. Filtering, and vacuum drying to obtain H₂TMPPSNa(TMPPS)。

③ taking TMPPS 342.6mg (0.3mmol) and 201.6mg (0.3mmol) of iridium chloride (1, 5-cyclooctadiene iridium chloride (I) dimer, [ Ir [)^ⅠCl(COD)]₂) The reaction is carried out in 30mL of glycol, heated in an oil bath at 140 ℃, magnetically stirred and condensed and refluxed. And (5) reacting for 5h, performing ultraviolet tracing reaction, wherein two groups of UV curves separated by 0.5h have no obvious difference, and confirming that the reaction is finished.

Fourthly, transferring the product after the reaction to a reduced pressure distillation device, vacuumizing by using a water pump, and distilling off the solvent ethylene glycol at 130 ℃. Dissolving the residual solid with appropriate amount of distilled water, filtering to remove some insoluble substances generated in the reaction, and washing twice with deionized water. The filtrate was collected and concentrated by heating. Dialyzing with dialysis bag with molecular weight cutoff of 1000Da to remove excessive iridium salt. Changing water every 30min, and dialyzing for 6 h. Vacuum distilling, and evaporating solvent water at 70 deg.C to obtain purple solid (IrTMPPS) with yield of 68%.¹H NMR(500MHz,DMSO-d⁶)9.31–8.76(m,8H,β-pyrroles),8.75–8.48(m,4H,2-phenyl),8.21–7.98(m,4H,6-phenyl),7.38(d,J＝26.7Hz,4H,5-phenyl),4.05(s,12H,-OCH₃)。

Example 2

A triphenylphosphine-porphyrin-iridium complex has a synthetic scheme shown in figure 2, and comprises the following preparation steps:

taking TPP (tetraphenylporphyrin, 4.00g, 6.5mmol) and trioxymethylene (0.1351g, 1.5mmol), a proper amount of hydrochloric acid, anhydrous zinc chloride and acetone, mixing and placing in a 150mL round-bottom flask, and keeping the temperature at 70 ℃ for 3 hours. After the reaction, the mixture was washed with deionized water, and the organic phase was retained by liquid separation. And (4) carrying out rotary evaporation on the organic phase to remove acetone, and carrying out column chromatography separation and purification on the solid to obtain the compound a.

② taking a and 40.0mL (98.0% H)₂SO₄) Ground into a uniform paste in a mortar. The paste was transferred to a 250mL round bottom flask, then 60mL 98% H was added₂SO₄And the mixture is heated in an oil bath at 90-120 ℃ for 6h with stirring. The mixture was placed in a 400mL beaker, cooled to room temperature, and then 300mL of deionized water was slowly added with stirring. After cooling, the solution was filtered through a G5 sintered glass filter to remove unreacted a, and 200mL of acetone was added to the filtrate, followed by stirring to precipitate a precipitate. The resulting precipitate was washed 2 times with acetone. Acetone was removed and the solid product was dissolved in 150mL deionized water and saturated NaHCO₃And (4) neutralizing. After the pH value is adjusted to 6.0, the solution is dialyzed and concentrated, and then is recrystallized twice by using a dehydrated methanol/ethanol mixed solvent. After filtration, vacuum drying, compound b is obtained.

③ taking b and triphenylphosphine (PPh3) (n (b): 1: 4.3) to mix and put into a single-neck round-bottom flask, and heating and refluxing for 12h at the temperature of 100 ℃ and 130 ℃ by taking toluene or 1, 4-dioxane as a solvent. And (3) obtaining a crude product of the c, removing the solvent, dissolving with deionized water, filtering to obtain filtrate, removing water, washing with diethyl ether, and drying to obtain a relatively pure compound c.

Fourthly, taking c and [ Ir^Ⅰ(COD)Cl]₂Mixing (n (c): n (Ir): 1), reacting in 30mL of ethylene glycol, heating in 140 ℃ oil bath, magnetically stirring, condensing and refluxing. And (3) carrying out reaction for 5h, carrying out ultraviolet tracing reaction, wherein two groups of UV curves separated by 0.5h have no obvious difference, changing the reaction device into a reduced pressure distillation device after the completion of the reaction is confirmed, vacuumizing by using a water pump, and evaporating out the solvent ethylene glycol at 130 ℃. Dissolving the residual solid with appropriate amount of distilled water, filtering to remove some insoluble substances generated in the reaction, and washing twice with deionized water. The filtrate was collected and concentrated by heating. Dialyzing with dialysis bag with molecular weight cutoff of 1000Da to remove excessive iridium salt. Changing water every 30min, and dialyzing for 6 h. Vacuum distilling, evaporating solvent water at 70 deg.C to obtain triphenylphosphine-porphyrin-iridium complexCompound, yield 48%.

¹H NMR(300M,DMSO-d⁶)7.70(4H,benzene),7.68(4H,benzene),7.46(4H,benzene),7.41(2H,pyrrole),7.36-7.33(60H,benzene),7.07(2H,ethylene),6.44(2H,ethylene),6.24(2H,ethylene),4.2(4H,-OH),2.6(8H,methylene)。

¹³C NMR(300M,DMSO-d⁶)143.6,139.5,139.2,138.6,138.6,138.0,137.0,135.8,135.0,134.9,130.5,130.4,129.9,124.9,123.8,122.8,121.2,120.2,120.1,115.4,29.6。

m/z:2258.41(100.0％),2257.41(77.0％),2256.41(59.5％),2255.40(45.8％),2259.41(32.9％),2259.41(31.4％),2260.42(22.1％),2257.41(19.6％),2257.41(18.7％),2260.41(18.1％),2259.40(13.9％),2258.41(13.1％),2258.40(10.8％),2257.40(8.3％),2261.42(8.1％),2261.41(6.0％),2261.41(5.7％),2260.42(5.3％),2259.42(4.8％),2262.41(4.0％),2259.41(3.5％),2259.41(3.4％),2259.41(3.2％),2258.41(3.1％),2260.41(2.9％),2258.41(2.5％),2260.41(2.4％),2259.41(2.2％),2262.42(2.1％),2257.41(1.9％),2258.41(1.7％),2259.41(1.5％),2256.40(1.5％),2263.42(1.5％),2257.41(1.3％),2260.42(1.3％),2262.40(1.2％),2258.40(1.1％),2259.42(1.1％),2260.41(1.1％),2260.41(1.0％)

Example 3

A methoxy-phthalocyanyl-iridium complex has a synthetic scheme shown in figure 3, and comprises the following preparation steps:

putting 4-nitro-1, 2-benzene dinitrile (6.6g, 38mmol) into a 150mL round-bottom flask, adding methanol (3.6mL, 114mmol) and K₂CO₃(10.4g, 76mmol, 2-fold equivalent), DMF (12 mL). The flask was stirred at room temperature for 72h, during which time the flask was wrapped with aluminum foil to keep the system in the dark. After 72h, the reaction mixture was transferred to a beaker containing 50mL of deionized water, the system had a yellow precipitate, and the suspension was stirred at room temperature for an additional 5 min. Subsequently, the filtrate was removed and the solid was collected. The solid was washed with 50mL of 0 ℃ cold methanol. The solid was collected and washed with ethyl acetate: column chromatography was performed with n-hexane (v: v ═ 1: 2) as eluent. Collecting the product fraction, removing the solvent to obtain dry and pure 4-methoxy-1, 2-benzenedinitrile, compound d.

② taking 4-methoxy-1, 2-benzene dinitrile (15.45mmol, 2.4437g) to put in a 50mL three-neck flask, adding 20mL n-amyl alcohol, pumping out air and introducing argon, refluxing and reacting at 135 ℃ for 2.5h, and simultaneously tracking the reaction progress by UV and TLC until the reaction is finished. And after the reaction is finished, carrying out reduced pressure distillation to remove n-amyl alcohol, cooling the product to room temperature, adding methanol, standing, precipitating, carrying out suction filtration, taking a filter cake, and purifying by taking dichloromethane/methanol as an elution system to obtain the hollow phthalocyanine (compound e).

③ hollow phthalocyanine e and 40.0mL (98.0% H)₂SO₄) Ground into a uniform paste in a mortar. The paste was transferred to a 250mL round bottom flask, then 60mL 98% H was added₂SO₄And the mixture is heated in an oil bath at 90-120 ℃ for 6h with stirring. After the reaction is finished, cooling to room temperature, and performing Soxhlet extraction with deionized water to obtain an extracting solution. NaHCO for extracting solution₃After neutralization to pH 6.0, the solution is dialyzed and concentrated, recrystallized for 2 times by using a dehydrated methanol/ethanol mixed solvent, filtered and dried to obtain a compound f.

Fourthly, taking f and [ Ir with equal molar quantity^Ⅰ(COD)Cl]₂Mixing, adding 40mL of xylene, N₂Reacting for 18h at 155 ℃ in the atmosphere to obtain the methoxy-phthalocyanine-iridium complex. The crude product was distilled off under reduced pressure to remove the solvent and purified by eluting with a dichloromethane/methanol system to give the product in 45% yield.

¹H NMR(300M,DMSO-d⁶)8.75(2H,benzylidenimin),8.42(1H,benzylidenimin),8.24(1H,benzene),8.02(2H,benzylidenimin),7.72(1H,benzene),7.63(1H,benzene),4.20(4H,-OH),3.83(16H,methyl)。

13C NMR(300M,DMSO-d6)171.4,169.7,168.0,161.1,159.4,137.7,137.7,135.0,134.3,132.6,126.2,125.0,124.1,122.2,114.7,114.4,112.5,108.3,106.6,100.2,55.8。

m/z:294.24(100.0％),293.74(59.5％),294.49(38.9％),293.99(23.2％),294.74(18.1％),294.24(10.8％),294.99(7.0％),294.75(4.7％),294.24(4.4％),294.49(4.2％),294.74(3.7％),294.49(3.2％),294.49(3.0％),294.75(2.7％),294.24(2.2％),293.99(1.9％),293.99(1.8％),295.00(1.4％),294.74(1.2％),295.24(1.2％),294.74(1.2％)。

Further, in order to verify the advancement of the methoxy-porphyrin-iridium complex prepared in example 1 of the present application, the present application examined the performance of the methoxy-porphyrin-iridium complex prepared in example 1.

Test example 1 uv/fluorescence/ir spectroscopy experiment:

the optical properties of the porphyrin-iridium complex prepared in example 1 can be preliminarily characterized by using an ultraviolet spectrum and a fluorescence spectrum, and the absorption peak region and the fluorescence emission peak position of the porphyrin-iridium complex are determined, and the test result is shown in fig. 4, wherein the abscissa is the wavelength, the left ordinate is the ultraviolet visible absorbance, and the right ordinate is the fluorescence excitation intensity. As can be seen from the attached figure 4, the porphyrin-iridium complex has a strong absorption band at 367-462 nm, namely an S band, and has a weaker absorption band at 506-543 nm, namely a Q band. From the fluorescence spectrogram, weak red phosphorescence emission can be seen at 636-740 nm.

To further refine the structural characterization, the infrared spectrum of the complex powder was determined by KBr tabletting method, as shown in fig. 5 (wave number on abscissa and transmittance on ordinate) and shown in table 1 below:

TABLE 1

From the above test results, it can be seen that the methoxy-porphyrin-iridium complex prepared in example 1 also has 3400cm in length because the two axial ligands of iridium ion are water molecules^-1An O-H stretching vibration peak is generated nearby. At-1600 cm^-1，～1500cm^-1And-1450 cm^-1The vibration peak of the skeleton of the benzene ring is nearby. In addition, due to the presence of methoxy groups therein, in the range of-1250 cm^-1And the nearby part also has a characteristic peak of the phO-C stretching vibration.

In conclusion, the methoxy-porphyrin-iridium complex prepared in example 1 is comprehensively and qualitatively analyzed in the modes of nuclear magnetism, ultraviolet light, infrared light, fluorescence and the like.

Test example 2 cytotoxicity test:

the application researches the cytotoxicity and dark toxicity of the methoxy-porphyrin-iridium complex to a liver cancer cell line (Hep-G2) through an MTT (thiazole blue) experiment. The method comprises the following specific steps:

(ii) seeding Hep-G2 cells in 96-well plates (5X 10)³Each well) was incubated at 37 ℃ for 24 hours for adherence, the old medium was removed, incubated with methoxy-porphyrin-iridium complex for 4 hours, and after incubation, the drug-containing medium was replaced with fresh medium containing no methoxy-porphyrin-iridium complex.

② the illumination group is 6.5mW/cm²The LED lamp (HTLD-S20-100 × 130,465nm) is illuminated for 10min, 20min and 30 min; the dark control group was not treated at all. The 96-well plate is placed back into the constant temperature incubator after the illumination of the illumination group, and the temperature is controlled at 37 ℃/5% CO₂Incubate for 44 h.

③ MTT (20. mu.L/well, 5mg/mL) was dissolved in PBS (phosphate buffered saline) and added to a 96-well plate for incubation for 4 h. The plates were removed of the medium containing MTT. DMSO (dimethyl sulfoxide, 100. mu.L/well) is added to a 96-well plate, the plate is uniformly shaken for 3min, after formazan crystals are completely and uniformly dissolved in DMSO, the 96-well plate is placed in a microplate reader, and the absorbance of each well on the plate at 490nm is read.

And fourthly, using cisplatin which is clinically applied as a positive control group and using blank holes as a negative control group.

Fifthly, after the methoxy-porphyrin-iridium complex and the cisplatin are administrated to the Hep-G2 cell line for 48 hours, the IC of each complex is calculated₅₀The values (half inhibitory concentrations) are shown in table 2 below.

TABLE 2

a represents the time of illumination under a 465nm LED lamp after the cells are incubated for 4h by the compound: 10min (3.9J/cm)²)，20min(7.8J/cm²) And 30min (11.7J/cm)²). b indicates that the cells were not treated with light after 4h incubation with compound.

From the above tests, it can be seen that the methoxy-porphyrin-iridium complex prepared in example 1 has an IC under dark conditions₅₀>50 μ M, indicating that the complex has no dark toxicity; under the condition of illumination, the methoxyl-porphyrin-iridium complex shows IC along with the increase of light dose₅₀The decreased value indicates an increased toxicity of the methoxy-porphyrin-iridium complex. The exposure to light for 10min alone showed higher toxicity than the cisplatin control. The toxicity of 20min illumination is not much different from that of 30min illumination, so 7.8J/cm is selected²Is a suitable light dose. At such low light doses, three times more toxicity than cisplatin was exhibited. The methoxy-porphyrin-iridium complex is expected to become a novel high-efficiency controllable antitumor drug.

Test example 3 cellular phosphorescence imaging experiment:

phosphorescence imaging is a biological non-destructive analysis technique that is widely used in biology and basic medical research. The application co-stained with methoxy-porphyrin-iridium complex prepared in example 1 using a commercial mitochondrial green dye (MTG), a commercial lysosomal green dye (LTG).

Hep-G2 cells were incubated in 10 μ M methoxy-porphyrin-iridium complex medium solution for 4h, then incubated with MTG (200nM,45min) and LTG (5 μ M,45min), respectively, and images were collected under a confocal microscope (Zeiss LSM 880S), as shown in FIG. 6, wherein Brightfield is the topography under the Bright field of the cells, IrTMPPS is the distribution map of the IrTMPPS in the cells, LTG and MTG are the distribution maps of the LTG and MTG in the cells, respectively, and overlay is the topography of the cells after the registration of the IrTMPPS with the LTG or MTG. It is understood that the luminescence site of the methoxy-porphyrin-iridium complex prepared in example 1 of the present application is substantially the same as that of LTG, indicating that the methoxy-porphyrin-iridium complex is substantially distributed in lysosomes of cells. Further, the co-dyeing coefficients of IrTMPPS and the superposition of LTG or MTG are analyzed by a correlation test chart, as shown in FIG. 7 and FIG. 8 respectively (the horizontal and vertical coordinates represent the light spot distribution of two color channels, respectively, and each occupies half of a quadrant by taking a diagonal as a center; when the light spots are superposed, the light spots are distributed to the diagonal): the overlapping line of the co-dyeing coefficient graph of IrTMPPS and LTG is in an oblique diagonal direction, which indicates that the high overlapping degree of two colors in the co-dyeing graph is achieved, and the co-dyeing coefficient (PCC, Pearson correlation coefficient) reaches 93 percent; the co-localization coefficient of IrTMPPS and MTG is only 36%, which indicates that the methoxy-porphyrin-iridium complex is localized in lysosomes of cells.

The above description is only exemplary of the present application and should not be taken as limiting the present application, as any modification, equivalent replacement, or improvement made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. An iridium complex is characterized in that the structural general formula of the iridium complex is shown as the following formula I or formula II:

2. The iridium complex of claim 1, wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈Each independently selected from: methoxy or triphenylphosphine methyl; alternatively, the first and second electrodes may be,

the R is₁、R₂、R₃、R₄Simultaneously selected from: methoxy or triphenylphosphine methyl; the R is₅、R₆、R₇、R₈Simultaneously selected from: methoxy or triphenyl radicalsA phosphonomethyl group; and/or the presence of a gas in the gas,

said Y is₁、Y₂Each independently selected from: sodium ion, potassium ion, ammonium ion, and chloride ion.

3. A preparation method of an iridium complex is characterized by comprising the following steps:

wherein, the structural general formula of the compound A is as follows:

or

4. The method for producing an iridium complex according to claim 3, wherein the sulfonation reaction conditions include: reacting for 5-8 hours at the temperature of 100-110 ℃; and/or the presence of a gas in the gas,

the step of neutralization treatment comprises: dissolving the compound B in water, adjusting the pH of an aqueous solution of the compound B to 6.0-7.0 by adopting a first alkaline salt, and separating to obtain a compound C; and/or the presence of a gas in the gas,

the step of coordination reaction comprises: and dissolving the compound C and the iridium precursor in the first organic solvent, and reacting for 5-6 hours at the temperature of 120-160 ℃.

5. The method for producing an iridium complex according to claim 4, wherein the volume ratio of the mass of the compound A to the concentrated sulfuric acid is 1 g: (20-30) ml; and/or

The molar ratio of the compound C to the iridium precursor is 1: (1-1.2); and/or

The alkaline salt is selected from: NH (NH)₄HCO₃、K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃At least one of; and/or

The iridium precursor is selected from: at least one of 1, 5-cyclooctadiene iridium (I) chloride dimer, chlorobis (cyclooctene) iridium (I) dimer, carbonylbis (triphenylphosphine) iridium (I) chloride; and/or

The first organic solvent is selected from: at least one of ethylene glycol, glycerol and dimethyl sulfoxide.

6. The method for producing an iridium complex according to claim 5, wherein the step of mixing the compound A with concentrated sulfuric acid and then conducting sulfonation comprises:

according to the volume ratio of the mass of the compound A to the concentrated sulfuric acid of 1 g: (8-15) ml, mixing and grinding the compound A and the concentrated sulfuric acid to obtain a pasty mixture;

adding the residual concentrated sulfuric acid into the pasty mixture, and reacting for 5-8 hours at the temperature of 100-110 ℃.

7. A preparation method of an iridium complex is characterized by comprising the following steps:

performing chloromethylation reaction on the compound D, and separating to obtain a compound E;

dissolving the compound G and triphenylphosphine in a second organic solvent, carrying out thermal reaction, and separating to obtain a compound H;

dissolving the compound H and an iridium precursor in a third organic solvent, carrying out a coordination reaction, and separating to obtain a second iridium complex grafted with triphenylphosphine methyl;

wherein the structural formula of the compound D is as follows:

or

8. The method for producing an iridium complex according to claim 7, wherein the step of subjecting the compound D to chloromethylation reaction comprises: dissolving the compound D, polyformaldehyde and hydrochloric acid in an organic solvent, reacting for 2-6 hours at the temperature of 60-100 ℃ under the condition of containing a chloride condensing agent, and separating to obtain a compound E; and/or the presence of a gas in the gas,

the conditions of the sulfonation reaction include: reacting for 5-8 hours at the temperature of 100-110 ℃; and/or the presence of a gas in the gas,

the step of neutralization treatment comprises: dissolving the compound F in water, adjusting the pH of an aqueous solution of the compound F to 6.0-7.0 by using the second alkaline salt, and separating to obtain a compound G; and/or the presence of a gas in the gas,

the conditions of the thermal reaction include: dissolving the compound G and triphenylphosphine in the second organic solvent, heating and refluxing for 12-24 hours, and separating to obtain a compound H; and/or the presence of a gas in the gas,

the step of coordination reaction comprises: and dissolving the compound H and the iridium precursor in the third organic solvent, and reacting for 12-24 hours at the temperature of 140-170 ℃.

9. The method for producing an iridium complex according to claim 8, wherein a molar ratio of the polyoxymethylene, the compound D, and the hydrochloric acid is 1: (4-5): (1-2); and/or the presence of a gas in the gas,

the volume ratio of the mass of the compound E to the concentrated sulfuric acid is 1 g: (20-30) ml; and/or the presence of a gas in the gas,

the molar ratio of the compound G to the triphenylphosphine was 1: (4-5); and/or the presence of a gas in the gas,

the molar ratio of the compound H to the iridium precursor is 1: (1-1.2); and/or the presence of a gas in the gas,

the polyoxymethylene is selected from: at least one of trioxymethylene, paraformaldehyde, formaldehyde dimethyl acetal, and chloromethyl ether; and/or the presence of a gas in the gas,

the chloride salt condensing agent is selected from: at least one of zinc chloride, aluminum chloride and tin chloride; and/or the presence of a gas in the gas,

the second basic salt is selected from: NH (NH)₄HCO₃、K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃At least one of; and/or the presence of a gas in the gas,

the iridium precursor is selected from: at least one of 1, 5-cyclooctadiene iridium (I) chloride dimer, chlorobis (cyclooctene) iridium (I) dimer, carbonylbis (triphenylphosphine) iridium (I) chloride; and/or the presence of a gas in the gas,

the second organic solvent is selected from: toluene and/or 1, 4-dioxane; and/or the presence of a gas in the gas,

the third organic solvent is selected from: at least one of ethylene glycol, xylene and dimethyl sulfoxide.

10. A photodynamic therapy medicament comprising an iridium complex as claimed in any one of claims 1 to 2 or an iridium complex produced by a process as claimed in any one of claims 3 to 9.