CN111407887B

CN111407887B - Cyclooxygenase-targeted near-infrared dye metal complex photosensitizer and preparation and application thereof

Info

Publication number: CN111407887B
Application number: CN201811554973.4A
Authority: CN
Inventors: 蔡林涛; 周理华; 向晶晶; 龚萍; 李洪锋
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2018-12-18
Filing date: 2018-12-18
Publication date: 2022-09-20
Anticipated expiration: 2038-12-18
Also published as: CN111407887A

Abstract

The invention provides a near-infrared dye-metal complex photosensitizer targeted by cyclooxygenase and its preparation and application. The structure of the photosensitizer of the present invention is shown in the following formula (I): M(L ₁ ) _m (L ₂ ) _n (X ₁ ) _o (I); wherein, M is a non-radioactive metal with an atomic number greater than 40; L ₁ is Neutral or anionic ligand; L ₂ has the following structure: R ₁ -L ₃ -R ₂ , wherein R ₁ is a cyclooxygenase targeting compound group, L ₃ is a linking group, and R ₂ is a near-infrared dye group; X ₁ is a counter ion; m, n are integers greater than or equal to 1 respectively; o is an integer from 0 to 4; the cyclooxygenase targeting compounds include: indomethacin and indomethacin derivatives; Infrared dyes include: rhodamine and rhodamine derivatives. The photosensitizer of the invention integrates the functions of tumor targeting, tumor imaging, drug tumor treatment and prognosis monitoring, and is a multifunctional organic small molecule photosensitizer.

Description

Cyclooxygenase-targeted near-infrared dye metal complex photosensitizer and preparation and application thereof

Technical Field

The invention relates to the field of photosensitizers, and particularly relates to an epoxidase-targeted near-infrared dye metal complex photosensitizer as well as preparation and application thereof.

Background

In 2008, the number of cancer deaths worldwide reaches 760 million (about 13% of all deaths). Nearly 1300 million cancer cases are newly diagnosed each year, and the number of deaths due to cancer is expected to reach 1310 ten thousand by 2030. Although regular screening and monitoring and early intervention are the best methods to improve survival, there is still a need to try to find better cancer treatment regimens, one that is both effective and efficient, acceptable and affordable to patients.

Chemotherapy, radiation therapy and surgical resection are the common cancer treatment options, chemotherapy, and recently small molecule-based therapies and immunotherapy combined with the above have emerged. However, chemotherapy often causes systemic side effects, surgical removal of tumors is associated with a high rate of recurrence, and the cumulative radiation dose limits the efficacy of radiation therapy.

Improvements to traditional medical protocols are important, but efforts should be made to develop safe, effective and cost-effective alternatives. Photodynamic therapy (PDT) is an alternative with tumor ablation and functional preservation. PDT was born in the beginning of the 20 th century and was first systematically described by Dougherty et al in 1975; since this time, PDT has been extensively studied and now PDT has become a specific disease treatment modality. In general, the process of PDT involves the use of a Photosensitizer (PS) that localizes the tumor, and then locally illuminating the tumor with light of a particular wavelength to activate the PS. And the excited PS then transfers its energy to molecular oxygen, thereby generating cytotoxic Reactive Oxygen Species (ROS), such as singlet oxygen (c ¹ O ₂ ) It can oxidize key macromolecules within the cell, leading to tumor cell ablation. In order not to damage adjacent normal tissues by ionizing light, which causes systemic toxicity and radiation therapy, unlike chemotherapeutic drugs, PDT uses 3 non-toxic components, which themselves have no toxic effect on biological systems. Indeed, PDT has its own advantages over conventional treatments, such as its minimal invasiveness, its repeated use without cumulative toxicity, excellent function and therapeutic effect, reduced long-term morbidity, and improved patient quality of life. PDT has been proven effective in the past four decades in superficial bladder cancer, early and obstructive lung cancer, esophageal cancer, head and neck cancer, skin cancer as well as it has been used as a post-operative adjuvant treatment to resect tumors to reduce residual tumor burden. Despite the increasing widespread use of PDT, PDT has not been achieved as a first-line tumor due to certain limitations (including lack of an ideal PS, challenges in formulating a PS, lack of dosimetry needed for complete and effective treatment, difficulty in formulating a treatment plan and monitoring treatment response)Intervention was clinically recognized.

An ideal PS would not only accumulate specifically in tumor tissue but also be rapidly cleared from normal tissue. Amphiphilic is also an ideal property that is necessary for PS because it requires some hydrophilicity for it to reach cancer tissues unimpeded and some hydrophobicity for it to bind to target cells when administered systemically. It must also be free of dark environment toxicity, high triplet particle yield, and it needs to have sufficient triplet particle survival time in order for the triplet particle to react sufficiently with ground state oxygen or other substrates to generate sufficient ROS. Just as it is required to satisfy so many different conditions, it is difficult to find or manufacture a pharmaceutical agent that can be called an ideal PS. However, some, if not all, of the drugs are used in clinical or clinical tests.

Researchers have never stopped exploring high performance photosensitizers for a long time. The development of photosensitizers goes through roughly three stages. The first phase photosensitizers were designed and synthesized primarily on the basis of porphyrins, which were developed in the early 70 and 80 th century. Such photosensitizers were the earliest and most clinically useful class of photosensitizers. They can kill cancer cells effectively and have low dark toxicity. However, their absorption coefficients are too small and often require the injection of large amounts of photosensitizer to achieve satisfactory results. In addition, the inability to selectively kill cancer cells is one of the major limitations for their further clinical use. The second type of photosensitizer mainly develops in the later 80's of the last century, and the photosensitizer overcomes the defect of weak absorbance of the first type of photosensitizer. The photosensitizer is mainly anthraquinone, phenothiazine, xanthene, cyanine and curcumin compounds. Such photosensitizers may achieve better cancer cell selectivity by adjusting the balance between hydrophilicity and lipophilicity. However, cancer cell selectivity still needs to be further improved. Since then, the focus of research in this area has been on how to increase the cancer cell selectivity of photosensitizers. The main measures are to connect a peptide chain, an antibody and other biological guide groups which are targeted for the specificity of the tumor to the existing photosensitizer or to include the photosensitizer in some nano-carriers or carriers with cancer cell selectivity to realize the targeted therapy of the cancer cells. These constitute a third class of photosensitizers. Such photosensitizers, while exhibiting some selectivity in vitro, have limited selectivity in vivo.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

The invention aims to provide a cyclooxygenase-targeted near-infrared dye metal complex photosensitizer, the complex provided by the invention not only has a tumor targeting effect, but also can enable near-infrared dye molecules to generate a triplet excited state through coordination with metal so as to realize diagnosis and treatment of tumors, and the photosensitizer is low in toxicity and high in selectivity.

The second purpose of the invention is to provide a preparation method of the cyclooxygenase-targeted near-infrared dye metal complex photosensitizer.

The third purpose of the invention is to provide the application of the cyclooxygenase-targeted near-infrared dye metal complex photosensitizer.

In order to achieve the above purpose of the present invention, the following technical solutions are adopted:

an oxidase-targeted near-infrared dye metal complex photosensitizer has a structure shown as the following formula (I): m (L) ₁ ) _m (L ₂ ) _n (X ₁ ) _o (I) (ii) a Wherein M is a nonradioactive metal with an atomic number greater than 40; l is ₁ Is a neutral or anionic ligand; l is ₂ The structure is as follows: r ₁ -L ₃ -R ₂ Wherein R is ₁ Is a cyclooxygenase-targeting compound, L ₃ Is a linking group, R ₂ Is a near infrared dye group; x ₁ Is a counter ion; m and n are integers greater than or equal to 1 respectively; o is an integer of 0 to 4; wherein the cyclooxygenase-targeting compound comprises: indomethacin and indomethacin derivatives; the near-infrared dye comprises: rhodamine and rhodamine derivatives, boron fluoride-complexed dipyrromethene dyes (bodipy dye), and malachite green bamboo green dyes (malachite gre)en) is used.

Meanwhile, the invention also provides a preparation method of the cyclooxygenase targeted near-infrared dye metal complex photosensitizer, which comprises the following steps: will (M) _p (L ₁ ) _q (X ₆ ) _r And contain L ₂ Reacting a ligand structure compound to obtain a cyclooxygenase-targeted near-infrared dye metal complex photosensitizer; wherein M is a nonradioactive metal with an atomic number greater than 40; l is ₁ Is a neutral or anionic ligand; l is ₂ The structure is as follows: r ₁ -L ₃ -R ₂ Wherein R is ₁ Is a cyclooxygenase-targeting compound, L ₃ Is a linking group, R ₂ Is a near infrared dye group; x ₆ Is a counter ion; p and q are integers which are more than or equal to 1 respectively; r is an integer of 0 or more; wherein the cyclooxygenase-targeting compound comprises: indomethacin and indomethacin derivatives; the near-infrared dye comprises: rhodamine and rhodamine derivatives, boron fluoride complex dipyrromethene dye (bodipy dye), and malachite green bamboo green dye (malachite green).

Furthermore, the invention also provides the application of the cyclooxygenase-targeted near-infrared dye metal complex photosensitizer in preparing a tumor imaging agent; and/or, the cyclooxygenase-targeted near-infrared dye metal complex photosensitizer is applied to the preparation of a tumor diagnostic agent; and/or the cyclooxygenase-targeted near-infrared dye metal complex photosensitizer is applied to the preparation of tumor therapeutic agents.

Compared with the prior art, the invention has the following beneficial effects:

the cyclooxygenase-targeted near-infrared dye metal complex photosensitizer provided by the invention integrates the functions of tumor targeting, tumor imaging, drug tumor treatment, prognosis monitoring and the like, is a multifunctional organic micromolecule photosensitizer, and has the following advantages:

the metal complex photosensitizer has the advantages of clear structure, stable construction, convenient synthesis method and easy control. Because the ligand structure has an indometacin structure with a tumor targeting effect, the targeting aggregation on a tumor part can be realized; meanwhile, the rhodamine group serving as the most preferable near-infrared dye molecule has the advantages of good light stability, large light absorption coefficient, high quantum yield, good water solubility and the like, active oxygen can be efficiently released to kill cancer cells after complexing with metal, and the rhodamine group has the effects of targeting, near-infrared fluorescence imaging and photodynamic therapy.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.

FIG. 1 is a reaction scheme of example 1;

FIG. 2 is a schematic diagram showing the Rho mass spectrometry detection of the intermediate of example 1;

FIG. 3 is a mass spectrometric detection of the intermediate Rho-COOH of example 1;

FIG. 4 is the intermediate IM-NH of example 1 ₂ Mass spectrometric detection;

FIG. 5 is a Rho-IM mass spectrometric view of the ligand of example 1;

FIG. 6 is a mass spectrometric detection of the product complex Ir-Rho-IM of example 1;

FIG. 7 is a reaction scheme of example 2;

FIG. 8 is a graph showing the comparison results of the metal-in-vitro photodynamic therapy experiment in Experimental example 1;

FIG. 9 is a graph showing the tumor targeting experiment results of the metal complex in Experimental example 2;

wherein, fig. 9(a) is a schematic diagram of in vivo fluorescence imaging distribution of the metal complex; (b) is a trend change chart of the fluorescence intensity of the metal complex living body; (c) the in vitro fluorescence imaging schematic diagram of various isolated organs and tumor tissues, and (d) the fluorescence intensity schematic diagram of various isolated organs and tumor tissues.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

In view of the defects of the existing photosensitizer in application, the invention provides a metal complex type photosensitizer compound, and by taking a near-infrared dye and a cyclooxygenase (especially cyclooxygenase-2, a substance is also a cancer cell enzyme) targeting compound as a ligand, the selectivity of the photosensitizer on cancer cells can be realized, and meanwhile, through the sum of bonds of the metal and the near-infrared dye, the near-infrared dye molecules can also generate a triplet excited state, so that the cancer cells are killed, and the dark toxicity is low.

In some embodiments of the invention, a cyclooxygenase-targeted near-infrared dye metal complex photosensitizer is provided having the structure shown in formula (I):

M(L ₁ ) _m (L ₂ ) _n (X ₁ ) _o (I)；

wherein M is a nonradioactive metal having an atomic number greater than 40; the metal is preferably one of Re, Ru, Os, Rh, Ir, Pd, Pt or Au, and is more preferably Ir;

ligand L ₁ The ligand is neutral or anionic ligand, and can be monodentate coordinating group or coordinating atom, or bidentate, tridentate, tetradentate, pentadentate, hexadentate coordinating group, etc.

Ligand L ₂ The structure is as follows: r ₁ -L ₃ -R ₂ ；

Wherein, ligand L ₂ In, R ₁ Is a cyclooxygenase-targeting compound, said cyclooxygenase-targeting compound comprising: indomethacin and indomethacin derivatives;

R ₂ is a near-infrared dye base, the near-infrared dye comprising: rhodamine and rhodamine derivatives, boron fluoride-complexed dipyrromethene dyes (bodipy dye), and malachite green bamboo green dyes(malachite green); preferably a rhodamine derivative.

L ₃ As the linking group, a functional group (via the functional group and R) at both ends may be mentioned ₁ 、R ₂ Bond and linkage) or a C1-C6 linear or branched alkylene group (methylene, ethylene, propylene, isopropylene, butylene, pentylene, hexylene, etc.), or a C1-C6 linear or branched heteroalkylene group (cysteamine, etc.) with a heteroatom (O, S, N, etc., preferably a disulfide bond);

the straight chain or branched chain alkyl or the straight chain or branched chain heteroalkyl can be further substituted by C1-C5 alkyl and alkoxy.

X ₁ The metal complex photosensitizer provided by the invention can be a neutral metal complex or an ionic metal complex as a counter ion;

when the metal complex photosensitizer provided by the invention is an ionic complex (the central metal atom of the complex cannot reach charge balance through a coordination group), a balance (anion) ion exists in the complex provided by the invention, and the balance ion can be a halogen ion, such as Cl ^- 、Br ^- 、I ^- Etc. may be PF ₄ -a plasma radical;

m and n are integers greater than or equal to 1 respectively;

when m and n are more than 1, different L ₁ 、L ₂ Groups which may be independently of the same or different structures;

o is an integer of 0 to 4;

the metal complex photosensitizer provided by the invention at least comprises one L ₁ And one L ₂ Ligands, and by the central metal with the appropriate amount L ₁ 、L ₂ Coordination, such that the metal complex has a stable coordination structure, e.g., when the central metal is Ir, a hexa-coordination structure needs to be achieved; when the central metal is Pd ²⁺ 、Pt ²⁺ Then four coordination is often required to achieve a stable structure.

Similarly, o is also chosen such that the overall complex remains charge balanced to achieve a stable structure (e.g., o is 0 if the complex is a neutral complex overall; if the complex is still positive after all ligands have been coordinated, an appropriate number of counterions is required to maintain charge balance).

In some preferred embodiments of the invention, the ligand L ₁ Includes a first ring and a second ring respectively bonded to the metal; wherein the first and second rings may be directly bonded or connected by an alkyl chain of C1-C3.

The first ring and the second ring are respectively and independently selected from one of imidazole, benzene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, oxazole, thiazole, oxadiazole, thiadiazole, furan or thiophene.

Wherein any hydrogen atom on the first and second rings is independently optionally substituted or unsubstituted;

when the hydrogen atom on the first ring and/or the second ring is substituted, the substituent may be one or more of an alkyl group of C1 to C6 (e.g., methyl, ethyl, propyl, isopropyl, butyl, pentyl, octyl, etc.), an aryl group of C5 to C12 (e.g., phenyl, biphenyl, etc.), (C1 to C4 alkylene) -C5 to C12 aryl group, C1 to C5 alkoxy group (methoxy, ethoxy, etc.), etc., and when the number of substituents is more than 1, different substituents may be independently the same or different.

Simultaneously, the first ring and the second ring are each independently optionally fused or non-fused to a third ring;

when the first ring and/or the second ring is fused with the optional third ring, a corresponding fused ring group, for example, one of naphthalene, quinoline, benzoxazole, benzofuran, benzopyridazine, benzopyrazinyl (quinoxaline), and the like, may be formed.

In some preferred embodiments of the invention, the ligand L ₁ Is an anionic ligand and comprises at least one metal bond and phenyl/substituted phenyl segment structure and one metal bond and heterocyclyl segment structure.

In some more preferred embodiments of the invention, ligand L ₁ The structure is shown as the following formula (II):

in the formula (II), R ₃ -R ₉ Each independently is any one of hydrogen, halogen (Cl, Br, I), substituted or unsubstituted straight or branched C1-C12 alkyl (such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl and the like), substituted or unsubstituted straight or branched C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy and the like), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl and the like), substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroaryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

the substituent of the substituted linear or branched C1-C12 alkyl group, C1-C12 heteroalkyl group, C1-C5 alkoxy group, C5-C12 aryl group, (C1-C4) alkylene-C5-C12 aryl group, C5-C12 heteroaryl group, (C1-C4) alkylene-C5-C12 heteroaryl group may be one or more of C1-C6 alkyl group (e.g., methyl, ethyl, propyl, isopropyl, butyl, pentyl, octyl, etc.), C5-C12 aryl group (e.g., phenyl, biphenyl, etc.), (C1-C4 alkylene) -C5-C12 aryl group, C1-C5 alkoxy group (methoxy, ethoxy, etc.), etc., and when the number of substituents is greater than 1, the different substituents may independently be the same or different.

At the same time, R ₃ -R ₉ Any adjacent R groups in (a) optionally form a saturated or unsaturated ring and form a fused ring with the attached phenyl/heterocyclic ring.

X ₂ Is N or C-R ₁₀ ；

Wherein, when X ₂ Is C-R ₁₀ When R is ₁₀ Is hydrogen, halogen, substituted or unsubstituted straight or branched C1-C12 alkyl (e.g., methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl, etc.), substituted or unsubstituted straight or branched C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy, etc.), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl, etc.)) Any of substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroaryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

Wherein, when X ₂ Is C-R ₁₀ When R is ₃ -R ₁₀ Any adjacent R group in (a) optionally forms a saturated or unsaturated ring and forms a fused ring with the attached benzene/heterocyclic ring, for example, a fused ring structure of naphthalene, quinoline, benzoxazole, benzofuran, benzopyridazine, benzopyrazine (quinoxaline), etc.

In still further preferred embodiments of the present invention, there are provided metal complex photosensitizers having the structure:

in the complex of the above formula (VI), each atom/group (substituent, L) ₂ Etc.), reference is made to the statements made above for the formulae (I), (II);

in particular, in the complex with the above structure, M is one of Re, Ru, Os, Rh, Ir, Pd, Pt or Au, and is preferably Ir.

In some preferred embodiments of the invention, L ₂ The structure is shown as the following formula (III):

R ₁₁ -R ₁₄ 、R ₁₆ -R ₂₆ each independently is any one of hydrogen, halogen (Cl, Br, I), substituted or unsubstituted straight or branched C1-C12 alkyl (such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl and the like), substituted or unsubstituted straight or branched C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy and the like), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl and the like), substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroaryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

At the same time, R ₁₆ -R ₂₆ Any adjacent R group in (a) optionally forms a saturated or unsaturated ring and forms a fused ring with the attached benzene/heterocyclic ring, for example, a fused ring structure of naphthalene, quinoline, benzoxazole, benzofuran, benzopyridazine, benzopyrazine (quinoxaline), etc.

R ₁₅ For bonding to the metal and the coordinating group, for example, any of phenylpyridine, bipyridine, quinoline, or isoquinolinyl may be used.

Wherein, any hydrogen atom on the phenylpyridine, bipyridine, quinoline, or isoquinolinyl group is independently and optionally substituted or unsubstituted;

when the above groups have substituents, the substituents may be one or more of C1-C6 alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, pentyl, octyl, etc.), C5-C12 aryl (e.g., phenyl, biphenyl, etc.), (C1-C4 alkylene) -C5-C12 aryl, C1-C5 alkoxy (methoxy, ethoxy, etc.), etc., and when the number of substituents is more than 1, different substituents may independently be the same or different.

X ₃ Is halogen (Cl, Br, I).

X ₄ 、X ₅ Are independently amide or ester;

L ₄ is C1-C12 alkyl (preferably C1-C5 alkyl such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, isopentyl, etc.), or C1-C12 heteroalkyl containing a disulfide bond (e.g., cystamine, etc.).

In some more preferred embodiments of the invention, L ₂ The structure is shown as the following formula (IV):

R ₁₁ -R ₁₄ 、R ₁₆ -R ₃₂ each independently is any one of hydrogen, halogen (Cl, Br, I), substituted or unsubstituted straight or branched C1-C12 alkyl (such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl and the like), substituted or unsubstituted straight or branched C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy and the like), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl and the like), substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroaryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

Wherein R is ₁₆ -R ₃₂ Any adjacent R group in (a) optionally forms a saturated or unsaturated ring and forms a fused ring with the attached benzene/heterocyclic ring, for example, a fused ring structure of naphthalene, quinoline, benzoxazole, benzofuran, benzopyridazine, benzopyrazine (quinoxaline), etc.

X ₃ Is halogen (Cl, Br, I).

X ₄ 、X ₅ Are respectively independent amide group or ester group.

in the complex of the above formula (VII), each atom/group (substituent, L) ₂ Etc.), reference is made to the statements made above for the formulae (I), (II), (III);

In some of the most preferred embodiments of the invention,

the structure of the near-infrared dye metal complex photosensitizer is shown as the following formula (V):

wherein, in the formula (V), M is one of Re, Ru, Os, Rh, Ir, Pd, Pt or Au, and is preferably Ir.

R ₃ -R ₉ Each independently is any one of hydrogen, halogen (Cl, Br, I), substituted or unsubstituted straight chain or branched C1-C12 alkyl (such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl and the like), substituted or unsubstituted straight chain or branched C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy and the like), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl and the like), substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroaryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

X ₂ Is N or C-R ₁₀ ；

Wherein, when X ₂ Is C-R ₁₀ When R is ₁₀ Is hydrogen, halogen, substituted or unsubstituted straight or branched chain C1-C12 alkyl (such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl and the like), substituted or unsubstituted straight or branched chain C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy and the like), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl and the like), substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroalkylAny of aryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

Wherein R is ₁₆ -R ₃₂ Any adjacent R group in (a) may optionally form a saturated or unsaturated ring, and form a fused ring with the benzene/heterocycle to which it is attached, for example, a fused ring structure in naphthalene, quinoline, benzoxazole, benzofuran, benzopyridazine, benzopyrazinyl (quinoxaline), and the like.

X ₄ 、X ₅ Are independently amide or ester;

L ₄ is C1-C12 alkyl (preferably C1-C5 alkyl such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, isopentyl, etc.), or C1-C12 heteroalkyl containing disulfide bonds (e.g., cystamine, etc.).

In some of the most preferred embodiments of the present invention, there are provided near-infrared dye metal complex photosensitizers having the following structure:

rhodamine has the advantages of good light stability, large absorptivity, high quantum yield, good water solubility and the like, and is widely applied to the aspects of biological probes, biological markers, environmental monitoring and the like. However, the research on rhodamine and its derivatives has basically focused on fluorescence imaging, and in order to apply rhodamine and its derivatives to photodynamic therapy, they must have triplet excited states under illumination conditions. However, rhodamine and its derivative triplet state and its report on photodynamic therapy are rare. The Detty, m.r. group imparts a triplet excited state to rhodamine by introducing S and Se atoms into the rhodamine system, but the dark toxicity of these compounds is so great that their further application in photodynamic therapy is hindered.

In the preferred complex provided by the invention, a rhodamine derivative group is coordinated with a metalThereby endowing rhodamine with a strategy of generating a triplet excited state, and the compounds can be efficiently generated under illumination ¹ O ₂ To kill the cells and their dark toxicity is small.

Further, the invention also provides a preparation method of the metal complex photosensitizer, which comprises the following steps:

will (M) _p (L ₁ ) _q (X ₆ ) _r And contain L ₂ Ligand structural compound (when L2 is neutral ligand, then directly react with compound L ₂ Reaction when L is ₂ When the ligand is anionic, it generally contains L ₂ As starting materials, e.g. HL ₂ Etc.) to obtain the cyclooxygenase targeted near-infrared dye metal complex photosensitizer;

wherein, M, L ₁ 、L ₂ Reference is made to the structures and definitions related to formulae (I), (II), (III) above;

meanwhile, p and q are integers which are more than or equal to 1 respectively; when q is greater than 1, different L ₁ May independently be the same or different groups;

r is an integer of 0 or more;

X ₆ the metal complex photosensitizer provided by the invention can be a neutral metal complex or an ionic metal complex as a counter ion;

when the metal complex photosensitizer provided by the invention is an ionic complex (the central metal atom of the complex cannot reach charge balance through a coordination group), a balance (anion) ion exists in the complex provided by the invention, and the balance ion can be a halogen ion, such as Cl ^- 、Br ^- 、I ^- Etc. may be PF ₄ ^- Plasma balancing the ionic groups.

In some preferred embodiments of the present invention, the metal complex photosensitizers of the present invention are prepared from M ₂ (X ₇ ) ₂ (L ₁ ) ₂ And contain L ₂ The compound with a group structure is obtained by reaction;

wherein, X ₇ Is halogen (Cl, Br, I), M, L ₁ 、L ₂ Is as defined in formula (I) above,

(II)、(III)。

In some more preferred embodiments of the present invention, the metal complex photosensitizers of the present invention are prepared from:

and

reacting to obtain;

wherein in formula VIII, X ═ is halogen (Cl, Br, I), and the remaining atoms or groups are as defined above with reference to formulae (I), (II), (VI); in formula (IX), the atoms or groups are as defined above with reference to formula (III).

In further preferred embodiments of the present invention, the metal complex photosensitizers of the present invention are prepared from:

and with

Reacting to obtain;

wherein, in formula VIII, X ═ is halogen (Cl, Br, I), and the remaining atoms or groups are defined as above with reference to formulae (I), (II), (VI); in formula (IX), the atoms or groups are as defined above with reference to formula (IV).

In a most preferred embodiment of the present invention, the metal complex photosensitizer of the present invention is prepared from:

and

and (3) reacting to obtain the compound.

Wherein, ligand L ₂ (R ₁ -L ₃ -R ₂ ) Can be prepared from R ₁ -X ₈ (i) AndX ₉ -L ₃ -X ₁₀ (ii) and R ₂ -X ₁₁ (iii) Reacting to obtain;

wherein R is ₁ 、R ₂ 、L ₃ For the definition of (A) and (B), refer to formula (I) above;

X ₉ 、X ₁₀ can be reacted with X ₈ Or X ₁₁ A bond and a functional group, preferably, X ₉ 、X ₁₀ Is amino, hydroxy, or carboxy, X ₈ And X ₁₁ Is a corresponding carboxyl, amino, or hydroxyl group.

In some preferred embodiments of the invention, L is present ₂ Ligand compound of structure consisting of

And X ₉ -L ₃ -X ₁₀ (ii) And an

Reacting to obtain;

for the definition of each compound group, reference is made to the compounds of formulae (II), (III) as above.

For example, compound (v) may be reacted with compound (ii) first and then with compound (iv) to give compound (iv) containing L ₂ A ligand compound of the structure.

In some more preferred embodiments of the invention, L is ₂ Ligand compound of structure consisting of

And

and X ₉ -L ₃ -X ₁₀ (ii) The preparation method comprises the following steps of.

For the definition of each compound group, reference is made to the compounds of formulae (II), (IV) as above.

In a particularly preferred mode of the invention, the ligand compound L ₂ Comprises the following steps:

and

and (3) reacting to obtain the compound.

In some embodiments of the invention, compound (v) may be represented by R ₃₃ CHO (vii) and

after reaction, the functional group is obtained through functional transformation;

wherein, in the compound (vii), R ₃₃ Is phenylpyridine, bipyridine, quinoline, or isoquinolinyl, each of which is optionally substituted or unsubstituted with at least one hydrogen atom independently, and at least one hydrogen atom is substituted with a methyl group that is functionally converted to form a corresponding amino, hydroxyl, or carboxyl group (preferably, the methyl group is oxidized with selenium dioxide to provide the corresponding carboxyl group);

R ₃₄ 、R ₃₅ Each independently is any one of hydrogen, halogen (Cl, Br, I), substituted or unsubstituted straight chain or branched C1-C12 alkyl (such as methyl, ethyl, propyl, butyl, isobutyl, pentyl, octyl, heptyl, dodecyl and the like), substituted or unsubstituted straight chain or branched C1-C12 heteroalkyl, substituted or unsubstituted C1-C5 alkoxy (methoxy, ethoxy, propoxy and the like), substituted or unsubstituted C5-C12 aryl (cyclopentadienyl, phenyl, naphthyl and the like), substituted or unsubstituted (C1-C4) alkylene-C5-C12 aryl, substituted or unsubstituted C5-C12 heteroaryl, or substituted or unsubstituted (C1-C4) alkylene-C5-C12 heteroaryl;

In a preferred embodiment of the invention, the compound (vi) may be prepared from

And with

After the reaction, the 5-position methyl function is converted (preferably, the methyl is oxidized into carboxyl);

for each compound group definition, reference is made to the compounds of formulae (IV), (viii) above.

In a particularly preferred embodiment of the invention, L ₂ Ligand compound

The method comprises the following steps:

the metal complex photosensitizer obtained by any method has good tumor cell enzyme (cyclooxygenase-2) targeting and tumor cell killing characteristics, so that the metal complex photosensitizer can be used for imaging, diagnosis and treatment of tumor cells.

Example 1

Indomethacin (100mg) and 1, 4-butanediamine-Boc (100mg) and the coupling reagents EDCI, HOBt, DMAP were added to a 25mL round-bottomed flask at room temperature, and after stirring rapidly for about 0.5 hour, the solution became clear,reacting for 24 hours at room temperature, stopping the reaction, removing the solvent by spinning, purifying by using a silica gel column, dissolving a sample by using dichloromethane, loading by using a wet method, and eluting by using dichloromethane: the gradient elution of methanol from 200:1 to 10:1, spin off the solvent and concentration gave IM-NHBoc. Yield 80%, IM-NHBoc deprotected under the action of hydrochloric acid to give IM-NH ₂ 。

Bpy (3.27g) was first dissolved in 1, 4-dioxane (150ml), charged to a 250ml round bottom flask and SeO was added at room temperature ₂ (2.174 g). Placing the mixture in N ₂ Reflux in a dark environment for 24 hours (gentle reflux, 100 ℃). The mixture was then filtered hot and the solution was cooled to room temperature, then the suspension was filtered, leaving the filtrate and the solvent removed. (rotary evaporator). The residue was dissolved in ethyl acetate (100 ml. times.3), and the suspension was filtered. Further using Na ₂ CO ₃ (1.0M, 100ML X2) washing the filtrate and Na ₂ S ₂ O ₅ The organic layer was washed (0.3M, 50ML X4) for 20 minutes each. And use Na ₂ CO ₃ The aqueous layer was adjusted to pH10 and supplemented with CH ₂ Cl ₂ (100ML X4) extraction of the solution followed by solvent removal gave bpy-CHO. A mixture of bpy-CHO (10mmol), 3- (diethylamino) phenol (3.4g), p-TsOH (0.258g) and acetic acid (100ml) was heated to 70 ℃ for 7 hours. The reaction was cooled to room temperature and the pH was adjusted to 7 with 10% NaOH solution. The precipitate was filtered and washed with water (100 ml). The solid was dissolved in DCM (100ml) and chloranil (1.23g) was added thereto. The mixture was stirred for 2 hours. After removal of the solvent, the residue was purified on column (from DCM/MeOH 100: 1) and bpy-Rho was obtained. Bpy-rho (1mmol), SeO ₂ (5mmol) of dioxane (10ml) was refluxed for 24 hours to give bpy-Rho-COOH.

In a 50ml round bottom flask, bpy-Rho-COOH (200mg) and IM-NH were added ₂ (200mg) and coupling reagents EDCI, HOBt, DMAP, reacting at room temperature for 24h, stopping the reaction, removing the solvent by spinning, purifying with silica gel column, dissolving the sample with dichloromethane, loading by wet method, eluting with dichloromethane: the gradient elution of methanol from 200:1 to 10:1, spin off the solvent and concentration gave Rho-IM with 25% yield. Taking 10mg of Rho-IM, and then adding 9mg of Ir ₂ (dpqx) ₄ -Cl ₂ Adding DCM and methanol into a 25ml round-bottom flask, adding 3ml of each of DCM and methanol, refluxing at 65 ℃ for 4h, adding KPF 626 mg, reacting at room temperature for 30min to generate a target product Ir-Rho-IM, and purifying.

Example 1 the reaction scheme is shown in figure 1 below.

Wherein the mass spectrum detection of intermediate Rho is shown in FIG. 2, the mass spectrum detection of intermediate Rho-COOH is shown in FIG. 3, and intermediate IM-NH ₂ The mass spectrum detection map is shown in FIG. 4, the ligand Rho-IM mass spectrum detection map is shown in FIG. 5, and the product complex Ir-Rho-IM mass spectrum detection map is shown in FIG. 6.

Example 2

With reference to the procedure of example 1, Rho-IM (rhodamine-indomethacin ligand) was prepared, followed by addition of 8mg of Ir ₂ (ppy) ₄ -Cl ₂ Adding DCM and methanol into a 25ml round-bottom flask, adding 3ml of each of DCM and methanol, refluxing at 65 ℃ for 4h, adding KPF 626 mg, reacting at room temperature for 30min, and purifying to obtain a target product Ir-Rho-IM.

Example 2 the reaction scheme is shown in figure 7.

Experimental example 1 experiment of photodynamic therapy outside metal-matched object

The product complex Ir-Rho-IM of example 1 and uncoordinated Rho-IM ligand were used as experimental materials, and the in vitro PDT treatment effect experiment was evaluated by CCK-8 analysis.

The specific experimental method comprises the following steps: under the dark condition, MCF-7 tumor cells are respectively incubated with rho-Ir-IM and rho-IM with different concentrations of 0, 0.5, 1, 2, 4, 7, 10 mu M and the like in a culture medium;

the group of culture media was irradiated with incandescent light at 11W for 30min, and then the survival rate of tumor cells in the group of culture media was measured, as shown in FIG. 8.

As can be seen from the results shown in FIG. 8, the survival rate of tumor cells gradually decreased with the increase in the concentration of rho-Ir-IM. The ligand rho-IM which is not chelated with the metal has little influence on the survival rate of the tumor cells, and the survival rate of the tumor cells can reach more than 80 percent even under the condition of high concentration of the ligand compound.

Experimental example 2 tumor targeting experiment with Metal Complex

The rho-Ir-IM product of example 1 is used as an experimental material, and the tumor targeting of the compound is tested by adopting an experimental method of living body near infrared fluorescence imaging (NIRF).

The constructed subcutaneous tumor-transplanted mouse model is injected with a metal complex rho-Ir-IM photosensitizer in the tail vein, and the change of the living fluorescence distribution and the fluorescence intensity trend is observed 48 hours after the injection, and the result is shown in figure 9.

From the NIRF image results of fig. 9a and b, the fluorescence intensity of rho-Ir-IM at the tumor site was steadily increased, and the highest peak was the strongest at 24 hours after i.v. injection.

The results of in vitro NIRF imaging of various ex vivo organs and tumor tissues after 24h injection are shown in fig. 9c, d. As can be seen from FIGS. 9c and d, the rho-Ir-IM of the present invention exhibits superior tumor tissue accumulation and relatively low uptake by various organs, further indicating that it has good targeting to tumor cells.

While particular embodiments of the present invention have been illustrated and described, it would be obvious that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.

Claims

1. the near-infrared dye-metal complex photosensitizer of a cyclooxygenase targeting is characterized in that, the structure of the near-infrared dye-metal complex photosensitizer of the cyclooxygenase targeting is shown in the following formula (I) :

M(L ₁ ) _m (L ₂ ) _n (X ₁ ) _o (I);

Among them, M is a non-radioactive metal with an atomic number greater than 40;

L ₁ is a neutral or anionic ligand;

The structure of L ₂ is as follows: R ₁ -L ₃ -R ₂ , wherein, R ₁ is a cyclooxygenase targeting compound group, L ₃ is a linking group, and R ₂ is a near-infrared dye group;

X ₁ is a counter ion;

m and n are integers greater than or equal to 1, respectively;

o is an integer from 0 to 4;

Wherein, the cyclooxygenase targeting compounds include: indomethacin and indomethacin derivatives;

The near-infrared dyes include: rhodamine and rhodamine derivatives;

The structure of L ₂ is shown in the following formula (III):

R ₁₁ -R ₁₄ , R ₁₈ , R ₂₀ -R ₂₃ , R ₂₅ -R ₂₆ are each independently hydrogen, halogen, substituted or unsubstituted linear or branched alkyl, substituted or unsubstituted linear or branched any of alkaneheteroalkyl, aryl, alkylenearyl, heteroaryl, or alkyleneheteroaryl;

R ₁₆ is any one of substituted or unsubstituted alkylene aryl or alkylene heteroaryl;

R ₁₇ , R ₁₉ , R ₂₄ are hydrogen, halogen, unsubstituted linear or branched alkyl, unsubstituted linear or branched heteroalkyl, aryl, alkylenearyl, heteroaryl, or any of alkylene heteroaryl;

R ₁₅ is any one of phenylpyridine, bipyridine, quinoline, or isoquinoline;

Wherein, any hydrogen atom on the phenylpyridine, bipyridine, quinoline, or isoquinoline group is independently optionally substituted or unsubstituted;

X ₃ is halogen;

X ₄ and X ₅ are each independently an amide group or an ester group;

L ₄ is an alkyl group, or a heteroalkyl group containing a disulfide bond.

2. The near-infrared dye-metal complex photosensitizer of cyclooxygenase targeting according to claim 1 is characterized in that, M is selected from among Re, Ru, Os, Rh, Ir, Pd, Pt, or Au. A sort of.

3. The near-infrared dye-metal complex photosensitizer of cyclooxygenase targeting according to claim 1, is characterized in that, L ₁ comprises the first ring and the second ring bonded with metal respectively;

The first ring and the second ring are independently selected from imidazole, benzene, pyridine, pyrimidine, pyrazine, pyridazine, pyrrole, oxazole, thiazole, oxadiazole, thiadiazole, furan, or thiophene. kind;

Wherein, any hydrogen atoms on the first ring and the second ring are independently optionally substituted or unsubstituted, and the first ring and the second ring are independently and optionally condensed with the third ring or non-dense.

4. The near-infrared dye-metal complex photosensitizer of cyclooxygenase targeting according to claim 3, is characterized in that, L ₁ structure is shown in following formula (II):

In formula (II), R ₃ -R ₉ are independently hydrogen, halogen, substituted or unsubstituted linear or branched alkyl, substituted or unsubstituted linear or branched heteroalkyl, aryl, any of alkylaryl, heteroaryl, or alkyleneheteroaryl;

Wherein, any adjacent R groups in R ₃ -R ₉ can optionally form a saturated or unsaturated ring;

X ₂ is N or CR ₁₀ ;

R ₁₀ is hydrogen, halogen, substituted or unsubstituted linear or branched alkyl, linear or branched heteroalkyl, alkoxy, aryl, alkylenearyl, heteroaryl, or alkylene any of the heteroaryl groups;

Wherein, when X ₂ is CR ₁₀ , any adjacent R groups in R ₃ to R ₁₀ can optionally form a saturated or unsaturated ring.

5. The near-infrared dye-metal complex photosensitizer of cyclooxygenase targeting according to claim 1, is characterized in that, L Structure is shown in following _formula (IV):

X ₃ is halogen;

X ₄ and X ₅ are each independently an amide group or an ester group;

L ₄ is an alkyl group, or a heteroalkyl group containing a disulfide bond.

6. The cyclooxygenase-targeted near-infrared dye-metal complex photosensitizer according to any one of claims 1-5, wherein the cyclooxygenase-targeted near-infrared dye-metal complex The photosensitizer structure is shown in the following formula (V):

Wherein, in formula (V), R ₃ -R ₉ are independently hydrogen, halogen, substituted or unsubstituted linear or branched alkyl, substituted or unsubstituted linear or branched heteroalkyl, aryl , any of alkylene aryl, heteroaryl, or alkylene heteroaryl;

X ₂ is N or CR ₁₀ ;

R ₁₀ is hydrogen, halogen, substituted or unsubstituted linear or branched alkyl, substituted or unsubstituted linear or branched heteroalkyl, aryl, alkylenearyl, heteroaryl, or alkylene any of the heteroaryl groups;

Wherein, when X ₂ is CR ₁₀ , any adjacent R groups in R ₃ -R ₁₀ can optionally form a saturated or unsaturated ring;

X ₃ is halogen;

X ₄ and X ₅ are each independently an amide group or an ester group;

L ₄ is an alkyl group, or a heteroalkyl group containing a disulfide bond.

7. The cyclooxygenase-targeted near-infrared dye-metal complex photosensitizer according to any one of claims 1-5, wherein the cyclooxygenase-targeted near-infrared dye-metal complexes The photosensitizer structure is shown below:

8. the preparation method of the near-infrared dye-metal complex photosensitizer of cyclooxygenase targeting according to any one of claims 1-7, is characterized in that, comprising:

(M) _p (L ₁ ) _q (X ₆ ) _r is reacted with a compound containing L ₂ ligand structure to obtain a near-infrared dye-metal complex photosensitizer targeted by cyclooxygenase;

Among them, M is a non-radioactive metal with an atomic number greater than 40;

L ₁ is a neutral or anionic ligand;

X ₆ is a counter ion;

p and q are integers greater than or equal to 1, respectively;

r is an integer greater than or equal to 0;

The near-infrared dyes include: rhodamine and rhodamine derivatives.

9. The application of the cyclooxygenase-targeted near-infrared dye-metal complex photosensitizer according to any one of claims 1-7 in the preparation of a tumor imaging agent;

And/or, the application of the cyclooxygenase-targeted near-infrared dye-metal complex photosensitizer in the preparation of a tumor diagnostic agent;

And/or, the application of the cyclooxygenase-targeted near-infrared dye-metal complex photosensitizer in the preparation of a tumor therapeutic agent.