Application of a derivative of hypocrellin with amino substitution or ethylenediamine substitution at the 2-position in the preparation of an anti-tumor photodynamic drug

Abstract

The present invention relates to a 2-amino-substituted derivative (Formula I) or an ethylenediamine-substituted derivative (Formulas II and III) of hypocrellin as a photodynamic drug for treating the following tumors: esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer cells related to digestive tract tumors; brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma cells related to head, neck and facial tumors; basal cell carcinoma, squamous cell carcinoma, cutaneous T-cell lymphoma, melanoma cells related to skin tumors; prostate cancer, bladder cancer cells related to genitourinary tumors. At the same time, such derivatives can localize the location of tumor tissues through their own fluorescence and are used for fluorescence-guided surgical resection of tumors.

Description

Technical Field

[0001] The present invention relates to the technical field of photosensitizer drugs, and particularly relates to the application of a hypocrellin derivative in the preparation of a photodynamic anti-tumor drug. Background Art

[0002] Photodynamic action is that under the action of light, a photosensitizer causes functional or morphological changes in body cells or biomolecules, thereby leading to cell damage and necrosis. The non-invasive or minimally invasive treatment technology that uses photodynamic action to selectively destroy diseased tissues to achieve the treatment purpose is called photodynamic therapy (PDT). In photodynamic therapy, the photosensitizer selectively accumulates in target cell tissues. When irradiated with light of an appropriate wavelength, it absorbs photon energy and changes from the ground state to the excited state. Since the excited photosensitive substance is extremely unstable, it will then release energy and return to the ground state through physical or chemical de-excitation processes. Among them, a large number of free radicals and singlet reactive oxygen species (ROS) are generated during the chemical de-excitation process. ROS act on cells to oxidize various biological macromolecules such as amino acids, unsaturated fatty acids, and adenosine, damage cell structures, affect cell functions, and thereby lead to cell death. However, the photosensitizer absorbed by normal tissues has been metabolized and excreted, without producing photodynamic action. The photosensitizer, light source, and oxygen are the three elements of photodynamic therapy. The key links of photodynamic therapy are that the photosensitizer specifically accumulates in the target, the light and the photosensitizer produce chemical reactions to generate reactive substances such as singlet oxygen, and the reactive substances directionally damage target cells.

[0003] Tumors are the most widely applied field of photodynamic therapy (PDT). Currently, in clinical practice, good results have been achieved in the PDT treatment of more than a dozen types of tumors. PDT can cure early-stage in-situ malignant tumors. For mid-stage tumors, it can be used for palliative treatment to improve symptoms and extend life. For PDT at the site of tumor surgical resection, laser irradiation can prevent tumor recurrence. Malignant tumors on the body surface are particularly suitable for PDT because the lesions are relatively shallow and the laser can directly penetrate the lesions. For malignant tumors in deeper locations, endoscopic light irradiation can be used for treatment. In addition, through some special methods, PDT can also be used for interventional treatment of liver cancer or bone marrow purification, etc. Compared with the three traditional treatment methods (surgery, chemotherapy, and radiotherapy), PDT is more targeted, can directly eliminate primary and recurrent tumors, has few side effects, rarely damages normal cells, and has a therapeutic effect on various types of tumors. With the improvement of photosensitizers and the development of PDT technology, the limitations and deficiencies of PDT are gradually being improved, but its clinical application still has certain limitations. For different patients, how to formulate individualized treatment plans, determine the treatment scope according to the size and depth of the tumor, as well as the selection of light sources and the magnitude of power, etc., all need further research. Currently, there are reports on the application of PDT in the treatment of tumors on the body surface, including skin tumors and precancerous lesions (such as basal cell carcinoma, squamous cell carcinoma, melanoma, cutaneous T-cell lymphoma, etc.), head and neck tumors (such as nasopharyngeal carcinoma, laryngeal carcinoma, tongue cancer, oral cancer, etc.), brain tumors (glioblastoma), reproductive tumors (such as prostate cancer, bladder cancer, cervical cancer, etc.), and digestive system tumors (such as cholangiocarcinoma, gastric cancer, lung cancer, liver cancer, colorectal cancer, etc.). PDT not only has good curative effects on many primary tumors, but also has unique technical advantages for many metastatic tumors. Research shows that the possible mechanisms of PDT in anti-tumor effects are direct killing of tumor cells or anti-vascular photodynamic therapy. Direct killing of tumor cells utilizes the dual selectivity of PDT. The photosensitizer selectively accumulates at the tumor cells, and under the irradiation of specific wavelength light waves, reactive oxygen species (ROS) are generated near the lesions, leading to the lesion and death of tumor cells. Anti-vascular photodynamic therapy is based on the fact that the survival of tumor cells depends on blood vessel supply. PDT can damage the vascular system related to tumors, resulting in ischemic death of tumors. If light irradiation is applied during the peak period of the photosensitizer concentration in the blood vessels, it can cause microvascular damage and insufficient blood supply in the lesions, thereby causing cell necrosis or apoptosis.

[0004] Photosensitizers are the most crucial factor in photodynamic therapy. The first-generation photosensitizers were developed in the 1970s and early 1980s. They were mainly porphyrin derivatives (HpD) represented by hematoporphyrin, which were mixtures extracted from pig or bovine blood. The main active ingredients were bishematoporphyrin ether or ester. It has the longest clinical application history and the most detailed research. This includes Carpoporphyrin, Photoporphyrin, and Photocarcinomycin independently developed in Canada, the United States, Germany, Russia, Belgium, and China. Over the past 30 years, there have been multiple commercial HpD products available for clinical use, and thousands of patients have received PDT treatment. Research results on HpD have achieved success in the treatment of some superficial tumors, and the treatment effect on respiratory tumors such as bronchogenic carcinoma is also quite remarkable. In addition, HpD also has a good therapeutic effect on upper gastrointestinal malignancies, and even a few cases have achieved clinical cure. Barrett's esophagus is a very important precancerous lesion of esophageal cancer, and currently HpD has become one of the preferred treatment methods. In addition, HpD also has a good therapeutic effect on head and neck tumors, brain tumors, bladder cancer, and cholangiocarcinoma. Although the first-generation photosensitizers have definite curative effects in tumor treatment, there are still many deficiencies, such as complex composition, poor tissue selectivity, slow metabolism, long light avoidance time, and certain toxicity. The development of the second-generation photosensitizers began in the late 1980s. The activity, absorption spectrum, and tissue selectivity of the second-generation photosensitizers have been greatly improved compared with the first-generation photosensitizers. Most of the second-generation photosensitizers are monomeric compounds, including porphyrin derivatives, metal phthalocyanines, chlorophyll degradation derivatives, polycyclic quinone compounds, and porphines. In 1990, the endogenous porphyrin photosensitizer ALA was successfully applied to the treatment of condyloma acuminata. Nowadays, ALA-mediated photodynamic therapy has been widely used in tumorous skin diseases such as squamous cell carcinoma and basal cell carcinoma; Hypericin photosensitizer has had many studies in anti-tumor in recent years. Hypericin-mediated PDT can treat various tumors such as pancreatic cancer, bladder cancer, lymphoma, prostate cancer, and basal cell carcinoma; Phthalocyanine photosensitizers have currently been applied in countries and regions such as Russia. Their anti-tumor and anti-infection effects are significant and good safety has also been shown in phase I / II clinical studies. The chemical structure of the second-generation photosensitizers is clear, the singlet oxygen yield is high, the photosensitive period is short, and the maximum absorption wavelength is red-shifted, increasing the depth of photodynamic therapy. Therefore, the commercialization and clinical application prospects are very optimistic, but there are still defects such as high difficulty in separation and purification and unsatisfactory targeting. The development of the third-generation photosensitizers began at the end of the 20th century. In order to improve biocompatibility, targeting, and develop a photosensitive drug delivery system, the developed third-generation photosensitizers are to combine porphyrin or phthalocyanine-based matrices with some chemical substances with biological properties such as amino acids, polymers, proteins, sugars, liposomes, antigens expressed by tumor tissues, antibodies or ligands corresponding to receptors, etc., to construct a photosensitive system that not only has tumor-directed localization but also can exert the effect of photodynamic therapy.For example, the combination of hematoporphyrin and monoclonal antibody has a very large killing effect on target cells; compared with the uncomplexed phthalocyanine complex, the uptake rate of the phthalocyanine-lipoprotein complex in tumor tissues has been greatly improved in both in vitro and in vivo experiments. At present, the third-generation photosensitizers are still in the preclinical animal research stage and are still some distance from true clinical application.

[0005] Although the above-mentioned photosensitizers have made great progress in the photodynamic therapy of tumors after decades of development in different periods, the types of such photosensitizers are relatively single, basically all derivatives of porphyrins. The metabolic time of porphyrin derivatives in the body is relatively long. The metabolic time of early HpD in the human body is 1-3 months. Although the metabolic time of later-developed sinoporphyrin sodium in the body has been greatly shortened to one or two weeks, there are still problems such as weak light absorption ability in the phototherapy window and difficulty in chemical separation of stereoisomers. Therefore, there is an urgent need to develop highly efficient photosensitizers with novel structures.

[0006] Hypocrellins are natural photosensitizers extracted and isolated from the stroma of the bamboo parasitic fungi Hypocrella bambusae and Shiraia bambusicola found in arrow bamboo forests above 3000 meters above sea level in Yunnan, China, and belong to perylenequinone compounds. Natural hypocrellins mainly include components such as hypocrellin A and hypocrellin B, among which hypocrellin A accounts for 95%. Under alkaline conditions, hypocrellin A can be dehydrated and converted into hypocrellin B, and the conversion rate can reach up to 99%. Compared with the phototherapy drug hematoporphyrin derivative (HpD) used clinically, hypocrellins have many advantages: such as simple composition, easy purification of raw materials, high triplet quantum yield and singlet oxygen quantum yield, high phototoxicity, low dark toxicity, both TypeⅠ and TypeⅡ dual photodynamic mechanisms, fast excretion, etc., and are a class of photosensitizers with very promising applications. In addition, the structure of hypocrellins is easy to be chemically modified, and the modified derivatives can meet the requirements that their absorption wavelength has strong light absorption ability in the phototherapy window (600-900nm) and their water solubility can meet the needs of clinical intravenous injection. Therefore, hypocrellins have broad application prospects as photodynamic drugs. However, the use of hypocrellin photosensitizers for the treatment of tumors is currently still in the laboratory research stage, and there are no related hypocrellin drugs for clinical research. Therefore, there is an urgent need to develop new hypocrellin-based photodynamic drugs for clinical photodynamic therapy of tumors.

[0007] The inventors first disclosed a 2-amino-substituted or ethylenediamine-substituted derivative of hypocrellin (CN201610894129.0). Through further research, we surprisingly found that such derivatives can efficiently kill certain specific tumor cells, such as esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, and colon cancer cells related to digestive tract tumors; head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, and glioblastoma cells related to head and neck tumors; basal cell carcinoma, squamous cell carcinoma, cutaneous T-cell lymphoma, and melanoma cells related to skin tumors; prostate cancer and bladder cancer cells related to genitourinary tumors. In addition, for the problem that there is no mediated drug to locate and guide the resection of tumors during the surgery for some special tumors (such as glioblastoma), the inventors found that such hypocrellin derivatives can specifically accumulate in glioblastoma cells and tissues, and there is no photosensitizer accumulation in the brain regions without tumors, showing good tumor-targeted enrichment. At this time, irradiating the tumor tissue with light of a specific wavelength can excite detectable fluorescence to locate the position of the tumor tissue, which can be used for fluorescence-guided surgery (FGS). Summary of the Invention

[0008] To solve the above technical problems, the present invention provides compounds represented by formula (I), formula (II), or formula (III), 2-amino-substituted or ethylenediamine-substituted derivatives of hypocrellin, their isomers, isotope-labeled substances, pharmaceutically acceptable salts, or solvates for use in the preparation of photodynamic anti-tumor drugs, wherein the tumors are esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell carcinoma, cutaneous T-cell lymphoma, melanoma, prostate cancer, and bladder cancer.

[0009]

[0010] In formula (I), R 1 has a structural general formula as shown in formula (IV), and R 2 is -H or -COCH 3 :

[0011]

[0012] In formula (IV), 0 ≤ m ≤ 12, 0 ≤ n ≤ 500, 0 ≤ p ≤ 12, 0 ≤ q ≤ 12; the m, n, p, q are zero or positive integers; Y is a linking group; Z is a terminal group; (OCH 2 CH 2 ) n is a polyethylene glycol unit;

[0013] In formula (IV), the linking group Y is O, NH, S, a carboxylic acid ester group, an amide group, a sulfocarboxylic acid ester group, a phenylene group, an alkenylene group having 3 to 12 carbon atoms or a cycloalkyl group having 3 to 12 carbon atoms;

[0014] The cycloalkyl group having 3 to 12 carbon atoms includes a substituted or unsubstituted cycloalkyl group or a cycloalkyl group containing a heteroatom, and the heteroatom is an oxygen, nitrogen or sulfur atom; the substituent is an alkyl group having 1 to 12 carbon atoms;

[0015] In formula (IV), the terminal group Z is hydrogen, an alkyl group having 1 to 12 carbon atoms, an alkoxy group having 1 to 12 carbon atoms, a phenyl group, a hydroxyl group, an amino group, a mercapto group, a carboxylic acid group, a sulfonic acid group, a pyridyl group, a quaternary ammonium salt or a pyridinium salt;

[0016] When the terminal group Z is a quaternary ammonium salt, the three substituents on the quaternary ammonium salt are each independently or simultaneously an alkyl group having 1 to 12 carbon atoms; the anion in the quaternary ammonium salt is an anion permitted in pharmaceutical preparations;

[0017] When the terminal group Z is a pyridinium salt, the substituents on the pyridine ring of the pyridinium salt are in the ortho, meta or para position; the pyridinium salt is formed by quaternizing pyridine with a halogenated hydrocarbon having 1 to 12 carbon atoms of different chain lengths; the anion in the pyridinium salt is an anion permitted in pharmaceutical preparations;

[0018] In formula (II), the substituent R 2 is -H or -COCH 3 :

[0019] In formula (II), R 3 ~R 8 are the same or different, as defined for the substituent R 1 in formula (I), that is, R 3 ~R 8 are the same or different and are independently as shown in formula (IV).

[0020] Preferably, in the above formula (IV), the linking group Y is: -O-; -NH-; -S-; -COO-; -O-CO-; -CONH-; -NH-CO-; -SO 3 -; -SO 2 -NH-; -C 6 H 4 -(phenyl); -C 3 H 4 -(cyclopropyl); -C 4 H 6 -(cyclobutyl); -C 5 H 8 -(cyclopentyl); -C 5 H 7 (CH 3 )-(methylcyclopentyl); -C 6 H10 -(cyclohexyl); -C 6 H 9 (CH 3 )-(methylcyclohexyl); -C 7 H 12 -(cycloheptyl); (piperazinyl).

[0021] Preferably, the terminal group Z in the above formula (IV) is: -H; -CH 3 ; -C 2 H 5 ; -C 4 H 9 ; -C 6 H 13 ; -OCH 3 ; -OC 2 H 5 ; -OC 4 H 9 ; -OC 6 H 13 ; -C 6 H 5 ; -OH, -NH 2 ; -SH; -COOH; -COOCH 3 ; -SO 3 H; -C 5 H 4 N; -C 5 H 4 N + ; -N + (CH 3 ) 3 ; -N + (C 2 H 5 ) 3 ; -N + (C 6 H 13 ) 3 ; -N + (CH 3 ) 2 (C 2 H 5 ) - N + (CH 3 ) 2 (C 6 H 13 ) - N + (CH 3 ) 2 (C 8 H 17 )

[0022] In one embodiment, m is an integer from 0 to 8, n is an integer from 0 to 200, p is an integer from 0 to 8, q is an integer from 0 to 8, such as 0, 1, 2, 3, 4, etc.

[0023] Specifically, the derivatives of Formula I and Formula I' are enol tautomers; the derivatives of Formula II and Formula II' are enol tautomers; the derivatives of Formula III and Formula III' are enol tautomers.

[0024]

[0025] Preferably, the substituent R 1 is alcohols with different chain lengths, and carboxylic acid esters formed by them and carboxyl polyethylene glycol: -(CH 2 ) m -OH; -(CH 2 ) m -OCH 3 ; -(CH 2 ) m -O-CO-CH 2 CH 2 -(OCH 2 CH 2 ) n -OCH 3 [m is an integer between 1 and 8, n is an integer between 0 and 100];

[0026] Preferably, the substituent R 1 is carboxylic acids with different chain lengths, and carboxylic acid esters or amides formed by them and polyethylene glycol: -(CH 2 ) m -COOH; -(CH 2 ) m -COOCH 3 ; -(CH 2 ) m -CO-(OCH 2 CH 2 ) n -OH; -(CH 2 ) m -CO-(OCH 2 CH 2 ) n -OCH 3 ; -(CH 2 ) m -CO-NH-CH 2 CH 2 -(OCH 2 CH 2 ) n -OCH 3[m is an integer between 1 and 8, and n is an integer between 0 and 100];

[0027] Preferably, the substituent R 1 is sulfonic acid groups with different chain lengths and sulfonic acid esters or sulfonamides formed by sulfonic acid groups and polyethylene glycol: -(CH 2 ) m -SO 3 H; -(CH 2 ) m -SO 2 -(OCH 2 CH 2 ) n -OH; -(CH 2 ) m -SO 2 -(OCH 2 CH 2 ) n -OCH 3 ; -(CH 2 ) m -SO 2 -NH-CH 2 CH 2 -(OCH 2 CH 2 ) n -OH; -(CH 2 ) m -SO 2 -NH-CH 2 CH 2 -(OCH 2 CH 2 ) n -OCH 3 [m is an integer between 1 and 8, and n is an integer between 0 and 100];

[0028] Preferably, the substituent R 1 is thia polyethylene glycol: -CH 2 CH 2 -SH; -CH 2 CH 2 -S-CH 2 CH 2 OH; -CH 2 CH 2 -S-CH 2 CH 2 OCH 3 ; -CH 2 CH 2 -S-CH 2 CH 2 -(OCH 2 CH 2) n -OH;

[0029] Preferably, the substituent R 1 is an alkyl group, an amino group, a hydroxyl group, or a substituent containing a phenyl group, a pyridyl group, or an alkene: -H; -CH 3 ; -C 2 H 5 ; -C 3 H 7 ; -C 4 H 9 ; -C 5 H 11 ; -C 6 H 13 ; -C 8 H 17 ; -NH 2 ; -NHCH 3 ; -NHC 2 H 5 ; -OH; -CH 2 CH=CH 2 ; -(CH 2 ) 2 CH=CH 2 ; -(CH 2 ) 3 CH=CH 2 ; -CH 2 C 6 H 5 ; -C 5 H 4 N; -CH 2 C 5 H 4 ; -(CH 2 ) 2 C 5 H 4 N; -NHC 6 H 5 ; -NHC 5 H 4 N;

[0030] Preferably, the substituent R 1 is a substituent containing a cycloalkyl group: -C 3 H 5 (cyclopropyl), -C 4 H 7 (cyclobutyl), -C 5 H 9 (cyclopentyl), -C 6 H 11 (cyclohexyl), -C 6 H 10 (CH 3)(methylcyclohexyl), -C 6 H 10 (OH)(hydroxycyclohexyl), -C 7 H 13 (cycloheptyl), -CH 2 C 6 H 10 COOH, -CH 2 C 6 H 10 COOCH 3 、-CH 2 C 6 H 10 OH, -C 6 H 10 COOH;

[0031] More preferably, the substituent R 1 is a cyclohexane with substituents (-C 6 H 10 -OH; -CH 2 C 6 H 10 COOH; -CH 2 C 6 H 10 COOCH 3 ; -CH 2 C 6 H 10 OH; -C 6 H 10 COOH), and the substituents are located at the ortho, para, and meta positions of the cyclohexane;

[0032] Preferably, the substituent R 1 is a substituent containing a quaternary ammonium salt: -(CH 2 ) m -N + (CH 3 ) 3 ; -(CH 2 ) m -N + (CH 3 ) 2 (C 2 H 5 ) - (CH 2 ) m -N + (CH 3 ) 2 (C 3 H 7 ) - (CH 2 ) m -N + (CH 3 )2 (C 4 H 9 );-(CH 2 ) 3 -N + (CH 3 ) 2 (C 6 H 13 );-(CH 2 ) m -N + (CH 3 ) 2 (C 8 H 17 );-(CH 2 ) m -N + (CH 3 ) 2 (C 12 H 25 );-(CH 2 ) m -O-CO-(CH 2 ) 2 -N + (CH 3 ) 3 ;-(CH 2 ) m -O-CO-(CH 2 ) 3 -N + (CH 3 ) 3 ;-(CH 2 ) m -O-CO-(CH 2 ) 4 -N + (CH 3 ) 3 ;-(CH 2 ) m -O-CO-(CH 2 ) 5 -N + (CH 3 ) 3 ;-(CH 2 ) m -O-CO-(CH 2 ) 6 -N + (CH 3 ) 3 ;-(CH 2 ) m -COO-(CH 2 ) 2 -N+ (CH 3 ) 3 ; -(CH 2 ) m -COO-(CH 2 ) 3 -N + (CH 3 ) 3 ; -(CH 2 ) m -COO-(CH 2 ) 4 -N + (CH 3 ) 3 ; -(CH 2 ) m -COO-(CH 2 ) 5 -N + (CH 3 ) 3 ; -(CH 2 ) m -COO-(CH 2 ) 6 -N + (CH 3 ) 3 ; -(CH 2 ) m -CONH-(CH 2 ) 2 -N + (CH 3 ) 3 ; -(CH 2 ) m -CONH-(CH 2 ) 3 -N + (CH 3 ) 3 ; -(CH 2 ) m -CONH-(CH 2 ) 4 -N + (CH 3 ) 3 [m is an integer between 1 and 8];

[0033] Preferably, the substituent R 1 is a heterocyclic-containing substituent:

[0034] According to an embodiment of the present invention, the tumor is esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell carcinoma of the skin, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer.

[0035] According to an embodiment of the present invention, the tumor cells are esophageal cancer cells AKR, gastric cancer cells MFC, lung cancer cells A549, liver cancer cells HepG2, cholangiocarcinoma cells MCC, colon cancer cells HCT116, head and neck cancer cells SCC2, brain cancer cells G442, tongue cancer cells TSCCa, nasal cancer cells KB, oral cancer cells CAL27, glioblastoma cells C6, basal cell carcinoma cells BCC, squamous cell carcinoma cells of the skin PECA, cutaneous T-cell lymphoma HH, melanoma cells B16, prostate cancer cells LNCaP, bladder cancer cells MBT-2.

[0036] According to an embodiment of the present invention, the drug can be a photodynamic drug, a fluorescence-mediated drug.

[0037] According to an embodiment of the present invention, the drug can be enriched in the tumor cells.

[0038] According to an embodiment of the present invention, the pharmaceutically acceptable salts include salts formed by the compound of formula (I) and an organic acid selected from propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid and citric acid or an acidic amino acid selected from aspartic acid and glutamic acid, followed by reaction with an inorganic base, including sodium, potassium, calcium, aluminum salts and ammonium salts, or salts formed by reaction with an organic base, including methylamine salts, ethylamine salts and ethanolamine salts; or salts formed by the compound of formula (I) and a basic amino acid selected from lysine, arginine and ornithine, followed by reaction with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid and phosphoric acid, or salts formed by reaction with an organic acid selected from formic acid, acetic acid, picric acid, methanesulfonic acid.

[0039] The present invention also provides the use of a compound represented by formula (I), formula (II) or formula (III), its isomers, isotope-labeled compounds, pharmaceutically acceptable salts or solvates in the preparation of a fluorescence-mediated drug for guiding the resection of the tumor boundary.

[0040] The present invention also provides the use of a compound represented by formula (I), formula (II) or formula (III), its isomers, isotope-labeled compounds or pharmaceutically acceptable salts in the treatment of tumor diseases, wherein the tumor is esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell carcinoma of the skin, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer.

[0041] According to an embodiment of the present invention, the treatment is photodynamic inactivation of tumor cells or fluorescence-mediated drug-guided resection of the tumor boundary.

[0042] The present invention also provides a method for preventing or treating tumor diseases, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of the compounds represented by formula (I), formula (II) or formula (III), their isomers, isotope-labeled compounds, pharmaceutically acceptable salts or solvates.

[0043] In some embodiments, the patient is a human.

[0044] Beneficial effects

[0045] 1) The compounds of formula (I), formula (II) or formula (III) of the present invention, as photodynamic drugs, can efficiently kill the following cancer or tumor cells: esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer. Such photodynamic drugs at a concentration of 20 - 30 nM can kill more than 90% of tumor cells, have basically no effect on normal cells, and the drug is basically excreted from the body after one week;

[0046] 2) The present invention for the first time discloses a hypocrellin derivative as a mediating drug in clinical tumor surgery to guide tumor resection. Such derivatives can specifically accumulate in tumor tissues, and there is no photosensitizer accumulation in areas without tumors, showing good tumor-targeted enrichment. At this time, when the tumor tissue is irradiated with light of a specific wavelength, detectable fluorescence can be excited to locate the position of the tumor tissue for fluorescence-guided tumor resection surgery. Brief description of the drawings

[0047] The following further details the specific embodiments of the present invention in conjunction with the drawings.

[0048] Figure 1 It is the general structural formula of hypocrellin 2-position amino-substituted or ethylenediamine-substituted derivatives.

[0049] Figure 2 It is the synthesis method of the derivative HB-4-PEGn of hypocrellin B with 2-position polyethylene glycol-amino pentanol substitution.

[0050] Figure 3 It is the reaction products HC-74 and HC-75 of deacylated hypocrellin HC and methyl ethylenediamine.

[0051] Figure 4 (a) is the absorption spectrum comparison chart of commercial porphyrin photosensitizers PpIX and Ce6. Figure 4(b) Absorption spectra comparison diagrams of hypocrellin B (HB), the 2-substituted product HB-6-PEG1 of HB (Example 6), and the ethylenediamine-substituted product HC-74 (Example 31).

[0052] Figure 5 ESR diagrams of the reaction of derivatives HB-45 and HC-45 (Example 23) with reactive oxygen species scavengers: (a) Singlet oxygen scavenger; (b) Superoxide radical scavenger.

[0053] Figure 6 (a) Photodegradation curve of HB-73 in Example 31; Figure 6 (b) Photodegradation curve of HC-73 in Example 31; Figure 6 (c) Photodegradation curve of HC-80 in Example 32.

[0054] Figure 7 Photostability comparison diagrams of derivatives HB-2-PEG4 (Example 2), HC-2-PEG8 (Example 2), HC-73 (Example 31), HC-80 (Example 32), and commercial photosensitizers (Ce6, hematoporphyrin HpD) under laser irradiation at 20 mW / cm 2 for 30 min.

[0055] Figure 8 pH stability comparison diagrams of derivatives HB-3-PEG4 (Example 3), HC-3-PEG8 (Example 3), HC-73 (Example 31), HB-77 (Example 32), and commercial photosensitizer (hematoporphyrin HpD).

[0056] Figure 9 Confocal fluorescence imaging diagrams of derivative HB-1-PEG12 (Example 1) in A549 cells: (a) Superimposed image of dark field and bright field; (b) Dark field image; (c) Bright field image.

[0057] Figure 10-1 Dark toxicity diagrams (a) and phototoxicity diagrams (b) of HB-1-PEG4 and HC-1-PEG8 (Example 1), and commercial photosensitizer HpD against esophageal cancer cells AKR.

[0058] Figure 10-2 Dark toxicity diagrams (a) and phototoxicity diagrams (b) of HB-3-PEG8 and HC-3-PEG16 (Example 3), and commercial photosensitizer HpD against lung cancer cells A549.

[0059] Figure 10-3 Dark toxicity diagrams (a) and phototoxicity diagrams (b) of HB-6-PEG2 and HC-6-PEG6 (Example 6), and commercial photosensitizer HpD against liver cancer cells HepG2.

[0060] Figure 10-4 Dark toxicity graphs (a) and phototoxicity graphs (b) of HB-14-PEG6 and HC-14-PEG12 (Example 13), and the commercial photosensitizer HpD against the colon cancer cell line HCT116.

[0061] Figure 10-5 Dark toxicity graphs (a) and phototoxicity graphs (b) of HB-19-PEG4 and HC-19-PEG8 (Example 15), and the commercial photosensitizer HpD against the cholangiocarcinoma cell line MCC.

[0062] Figure 10-6 Dark toxicity graphs (a) and phototoxicity graphs (b) of HB-45 and HC-45 (Example 23), and the commercial photosensitizer HpD against the gastric cancer cell line MFC.

[0063] Figure 11 (a) Phototoxicity graph of HB-10-PEG4 and HC-10-PEG8 (Example 10), and HpD against the brain cancer cell line G442;

[0064] Figure 11 (b) Phototoxicity graph of HB-12-PEG8 and HC-12-PEG12 (Example 12), HpD against the head and neck cancer cell line SCC2;

[0065] Figure 11 (c) Phototoxicity graph of HB-20-PEG6 and HC-20-PEG12 (Example 16) and HpD against the tongue cancer cell line TSCCa.

[0066] Figure 12 (a) Phototoxicity graph of HB-29 and HC-29 (Example 19), and the commercial photosensitizer HpD against the nasal cancer cell line KB;

[0067] Figure 12 (b) Phototoxicity graph of HB-61 and HC-61 (Example 28), and HpD against the oral cancer cell line CAL27;

[0068] Figure 12 (c) Phototoxicity graph of HB-73 (Example 31), HC-80 (Example 32), and HpD against the glioblastoma cell line C6.

[0069] Figure 13 (a) Phototoxicity graph of HB-4-PEG8 and HC-4-PEG16 (Example 4), HpD against the basal cell carcinoma cell line BCC;

[0070] Figure 13(b) Phototoxicity graphs of HB-9-PEG4 and HC-9-PEG8 (Example 9), and HpD against melanoma cell B16;

[0071] Figure 13 (c) Phototoxicity graphs of HB-36 and HC-36 (Example 20), and HpD against squamous skin cancer cell PECA.

[0072] Figure 14 (a) Phototoxicity graphs of HB-70 and HC-71 (Example 30), and HpD against prostate cancer cell LNCaP;

[0073] Figure 14 (b) Phototoxicity graphs of HB-73 and HC-73 (Example 31), and HpD against bladder cancer cell MBT-2;

[0074] Figure 14 (c) Phototoxicity graphs of HC-80 (Example 32) and HC-81 (Example 33), and agent HpD against cutaneous T-cell lymphoma cell HH.

[0075] Figure 15 Fluorescence imaging graphs in mice with different tumor models 4 h after tail vein injection of the derivatives of the present invention: (a) HB-1-PEG4 (Example 1) for imaging esophageal cancer tumor AKR cells; (b) HC-3-PEG12 (Example 3) for imaging gastric cancer MFC cells; (c) HB-6-PEG8 (Example 6) for imaging lung cancer A549 cells; (d) HB-7-PEG16 (Example 7) for imaging liver cancer HepG2 cells; (e) HC-9-PEG10 (Example 9) for imaging cholangiocarcinoma MCC cells; (f) HC-1-PEG50 (Example 1) for imaging colon cancer HCT116 cells.

[0076] Figure 16 Fluorescence imaging graphs in mice with different tumor models 4 h after tail vein injection of the derivatives of the present invention: (a) HB-11-PEG6 (Example 11) for imaging brain cancer G442 cells; (b) HC-18-PEG8 (Example 15) for imaging head and neck cancer SCC2 cells; (c) HC-45 (Example 23) for imaging tongue cancer TSCCa cells; (d) HC-57 (Example 27) for imaging oral cancer CAL27 cells; (e) HB-65 (Example 29) for imaging nasal cancer KB cells; (f) HC-81-PEG16 (Example 34) for imaging glioma C6 cells.

[0077] Figure 17Fluorescence imaging diagrams of the derivatives of the present invention in mice with different tumor models 4 h after tail vein injection: (a) HB-5-PEG10 (Example 5) for imaging basal cell carcinoma BCC cells; (b) HC-8-PEG12 (Example 8) for imaging squamous cell carcinoma PECA cells; (c) HB-64 (Example 28) for imaging melanoma B16 cell tumors; (d) HB-89-PEG16 (Example 36) for imaging cutaneous T cell lymphoma HH cells; (e) HC-90-PEG30 (Example 37) for imaging prostate cancer LNCaP cells; (f) HC-91-PEG16 (Example 38) for imaging bladder cancer MBT-2 cells.

[0078] Figure 18 Fluorescence imaging diagrams of the derivatives of the present invention in mice with different tumor models 4 h after intratumoral injection: (a) HB-28 (Example 19) for imaging basal cell carcinoma BCC cells; (b) HC-36 (Example 20) for imaging squamous cell carcinoma PECA cells; (c) HB-45 (Example 23) for imaging melanoma B16 cells; (d) HC-73 (Example 31) for imaging cutaneous T cell lymphoma HH cells; (e) HC-77 (Example 32) for imaging lung cancer A549 cells; (f) HC-80 (Example 32) for imaging cholangiocarcinoma MCC cells.

[0079] Figure 19 Fluorescence imaging diagrams of HB-3-PEG12 in Example 3 in tumor-bearing mice at 0-5 h after tail vein injection.

[0080] Figure 20 For a 635 nm laser with an intensity of 0.1 W / cm 2 Photodynamic therapy effect diagrams of mice inoculated with lung cancer cells (A549) 6 days and 12 days after photodynamic therapy after irradiation with a 635 nm laser with an intensity of 0.1 W / cm

[0081] Figure 21 After different photosensitizing drugs were administered by tail vein injection, with an intensity of 0.1 W / cm 2The tumor site pictures 6 days after photodynamic therapy of tumor-bearing mice inoculated with a 635 nm laser light for 10 min: Figure (a) shows a mouse inoculated with glioma cells (C6) without drug injection, only saline injection; Figure (b) shows a mouse inoculated with glioma cells (C6) with tail vein injection of drug HB-6-PEG6 (Example 6); Figure (c) shows a mouse inoculated with melanoma cells (B16) with tail vein injection of drug HC-9-PEG8 (Example 9); Figure (d) shows a mouse inoculated with bladder cancer cells (MBT-2) with tail vein injection of drug HB-63 (Example 28);

[0082] Figure 22 After different photosensitizing drugs were administered by intratumoral injection, the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 The tumor site pictures 6 days after photodynamic therapy of tumor-bearing mice inoculated with a 635 nm laser light for 10 min: Figure (a) shows a mouse inoculated with cholangiocarcinoma cells (MCC) with intratumoral injection of drug HB-29 (Example 19); Figure (b) shows a mouse inoculated with colon cancer cells (HCT116) with intratumoral injection of drug HB-45 (Example 23); Figure (c) shows a mouse inoculated with oral cancer cells (CAL27) with intratumoral injection of drug HC-73 (Example 31); Figure (d) shows a mouse inoculated with basal cell carcinoma cells (BCC) with intratumoral injection of drug HC-80 (Example 32);

[0083] Figure 23 Shows the photodynamic effect diagrams of derivatives HB-1-PEG6 (Example 1), HB-73 (Example 31), and HC-80 (Example 32) on HeLa cells. Detailed implementation manners

[0084] The technical solutions of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only for illustrative and explanatory purposes of the present invention, and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0085] Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products, or can be prepared by known methods.

[0086] In the present invention, the experimental methods are all conventional methods unless otherwise specified. The raw materials used can be obtained from public commercial channels unless otherwise specified; the percentages are all mass percentages unless otherwise specified; M is mol / L unless otherwise specified.

[0087] The raw materials used in the present invention are hypocrellin A (HA), hypocrellin B (HB), and deacetylhypocrellin C (HC); all the hypocrellin derivatives involved in the present invention are shown in Examples 1 to 38, and the characterization results of some derivatives are shown in Table 1.

[0088] In addition, it should be noted that the hypocrellin derivatives to be protected in the present invention all contain two enol tautomers, and the chemical structures of the two isomers are of course within the protection scope as shown in Formulas (I) and (I'), (II) and (II'), (III) and (III'). For the sake of simplicity, only one enol tautomer is listed in all the examples of the present invention, and the other enol tautomer and its corresponding general structural formula are described in detail in the specification, and its structure is of course within the protection scope. In addition, the general structural formula of the hypocrellin derivatives involved in the present invention contains a polyethylene glycol unit (PEGn), and the number of units n is any integer between 0 and 100, and the corresponding chemical structures are of course within the protection scope. For the sake of simplicity, only some integers are listed in all the examples of the present invention, and the corresponding general structural formulas of the rest are described in detail in the specification, and its structure is of course within the protection scope. Any range recorded in the present invention includes the end values and any numerical values between the end values and any sub-ranges constituted by the end values or any numerical values between the end values.

[0089] Example 1

[0090] Hypocrellin B (HB) or deacetylhypocrellin C (HC) reacts with aminoethanol, aminoethanol methyl ether, and aminoethanol-polyethylene glycol. The 2-position substitution products of HB are HB-1, HB-1-CH 3 , HB-1-PEGn, and the 2-position substitution products of HC are HC-1, HC-1-CH 3 , HC-1-PEGn (n is an integer between 1 and 100), and their structural formulas are shown as follows:

[0091]

[0092] Example 2

[0093] HB (or HC) reacts with aminopropanol, aminopropanol methyl ether, and aminopropanol-polyethylene glycol respectively. The 2-position substitution products of HB are HB-2, HB-2-CH 3 , HB-2-PEGn, and the 2-position substitution products of HC are HC-2, HC-2-CH 3 , HC-2-PEGn (n is an integer between 1 and 100), and their structural formulas are shown as follows:

[0094]

[0095] Example 3

[0096] HB (or HC) reacts with aminobutanol, aminobutanol methyl ether, and aminobutanol - polyethylene glycol respectively. The 2 - substituted products of HB are HB - 3, HB - 3 - CH 3 , HB - 3 - PEGn respectively, and the 2 - substituted products of HC are HC - 3, HC - 3 - CH 3 , HC - 3 - PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0097]

[0098] Example 4

[0099] HB (or HC) reacts with aminopentanol, aminopentanol methyl ether, and aminopentanol - polyethylene glycol respectively. The 2 - substituted products of HB are HB - 4, HB - 4 - CH 3 , HB - 4 - PEGn respectively, and the 2 - substituted products of HC are HC - 4, HC - 4 - CH 3 , HC - 4 - PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0100]

[0101] Example 5

[0102] HB (or HC) reacts with aminohexanol, aminohexanol methyl ether, and aminohexanol - polyethylene glycol respectively. The 2 - substituted products of HB are HB - 5, HB - 5 - CH 3 , HB - 5 - PEGn respectively, and the 2 - substituted products of HC are HC - 5, HC - 5 - CH 3 , HC - 5 - PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0103]

[0104] Example 6

[0105] HB (or HC) reacts with amino polyethylene glycol and amino polyethylene glycol methyl ether respectively. The 2 - substituted products of HB are HB - 6 - PEGn, HB - 6 - PEGn - CH 3 , and the 2 - substituted products of HC are HC - 6 - PEGn, HC - 6 - PEGn - CH 3 (n is an integer between 1 and 100), and their structural formulas are as follows:

[0106]

[0107] Example 7

[0108] HB (or HC) reacts with glycine, methyl glycinate, and glycine-polyethylene glycol respectively. The 2-position substitution products of HB are HB-7, HB-7-CH 3 , HB-7-PEGn, and the 2-position substitution products of HC are HC-7, HC-7-CH 3 , HC-7-PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0109]

[0110] Example 8

[0111] HB (or HC) reacts with alanine, methyl alaninate, and alanine-polyethylene glycol respectively. The 2-position substitution products of HB are HB-8, HB-8-CH 3 , HB-8-PEGn, and the 2-position substitution products of HC are HC-8, HC-8-CH 3 , HC-8-PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0112]

[0113] Example 9

[0114] HB (or HC) reacts with butyric acid, methyl butyrate, and butyric acid-polyethylene glycol respectively. The 2-position substitution products of HB are HB-9, HB-9-CH 3 , HB-9-PEGn, and the 2-position substitution products of HC are HC-9, HC-9-CH 3 , HC-9-PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0115]

[0116] Example 10

[0117] HB (or HC) reacts with valeric acid, methyl valerate, and valeric acid-polyethylene glycol respectively. The 2-position substitution products of HB are HB-10, HB-10-CH 3 , HB-10-PEGn, and the 2-position substitution products of HC are HC-10, HC-10-CH 3 , HC-10-PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0118]

[0119] Example 11

[0120] HB (or HC) reacts with 6 - aminocaproic acid, methyl 6 - aminocaproate, and 6 - aminocaproic acid - polyethylene glycol respectively, and the 2 - position substitution products of HB are HB - 11, HB - 11 - CH 3 , HB - 11 - PEGn respectively, and the 2 - position substitution products of HC are HC - 11, HC - 11 - CH 3 , HC - 11 - PEGn (n is an integer between 1 and 100), and their structural formulas are shown as follows:

[0121]

[0122] Example 12

[0123] HB (or HC) reacts with glycine amide - polyethylene glycol and alanine amide - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 12 - PEGn and HB - 13 - PEGn respectively, and the 2 - position substitution products of HC are HC - 12 - PEGn and HC - 13 - PEGn respectively (n is an integer between 1 and 100), and their structural formulas are shown as follows:

[0124]

[0125] Example 13

[0126] HB (or HC) reacts with butyramide - polyethylene glycol and valeramide - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 14 - PEGn and HB - 15 - PEGn respectively, and the 2 - position substitution products of HC are HC - 14 - PEGn and HC - 15 - PEGn respectively (n is an integer between 1 and 100), and their structural formulas are shown as follows:

[0127]

[0128] Example 14

[0129] HB (or HC) reacts with methanesulfonamide - polyethylene glycol and ethanesulfonamide - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 16 - PEGn and HB - 17 - PEGn respectively, and the 2 - position substitution products of HC are HC - 16 - PEGn and HC - 17 - PEGn respectively (n is an integer between 1 and 100), and the structural formulas are shown as follows:

[0130]

[0131] Example 15

[0132] HB (or HC) reacts with 3-aminopropanesulfonic acid-polyethylene glycol and 4-aminobutanesulfonic acid-polyethylene glycol respectively. The 2-position substituted products of HB are HB-18-PEGn and HB-19-PEGn respectively, and the 2-position substituted products of HC are HC-18-PEGn and HC-19-PEGn respectively (n is an integer between 1 and 100). The structural formulas are as follows:

[0133]

[0134] Example 16

[0135] HB (or HC) reacts with methanesulfonamide-polyethylene glycol and ethanesulfonamide-polyethylene glycol respectively. The 2-position substituted products of HB are HB-20-PEGn and HB-21-PEGn respectively, and the 2-position substituted products of HC are HC-20-PEGn and HC-21-PEGn respectively (n is an integer between 1 and 100). The structural formulas are as follows:

[0136]

[0137] Example 17

[0138] HB (or HC) reacts with 3-aminopropanesulfonamide-polyethylene glycol and 4-aminobutanesulfonamide-polyethylene glycol respectively. The 2-position substituted products of HB are HB-22-PEGn and HB-23-PEGn respectively, and the 2-position substituted products of HC are HC-22-PEGn and HC-23-PEGn respectively (n is an integer between 1 and 100). The structural formulas are as follows:

[0139]

[0140] Example 18

[0141] HB (or HC) reacts with 2-aminoethanethiol and 2-aminoethanethiolamine respectively. The 2-position substituted products of HB are HB-24, HB-24-CH 3 、HB-25, and the 2-position substituted products of HC are HC-24, HC-24-CH 3 、HC-25. The structural formulas are as follows:

[0142]

[0143] Example 19

[0144] HB (or HC) reacts with alkylamines with different chain lengths (ethylamine, propylamine, butylamine, hexylamine, octylamine) respectively. The 2-position substituted products of HB are HB-26 to HB-30, and the 2-position substituted products of HC are HC-26 to HC-30. The structural formulas are as follows:

[0145]

[0146] Example 20

[0147] HB (or HC) reacts with hydrazine, hydroxylamine, cyclopropylamine, cyclobutylamine, cyclopentylamine, and cyclohexylamine respectively. The 2-position substitution products of HB are HB-31 to HB-36 respectively, and the 2-position substitution products of HC are HC-31 to HC-36 respectively. Their structural formulas are as follows:

[0148]

[0149]

[0150] Example 21

[0151] HB (or HC) reacts with allylamine, butenylamine, hexenylamine, and octenylamine respectively. The 2-position substitution products of HB are HB-37 to HB-40 respectively, and the 2-position substitution products of HC are HC-37 to HC-40 respectively. Their structural formulas are as follows:

[0152]

[0153] Example 22

[0154] HB (or HC) reacts with benzylamine, aminomethylpyridine, aminobutylpyridine, and aminobutylpyridine salt respectively. The 2-position substitution products of HB are HB-41 to HB-44 respectively, and the 2-position substitution products of HC are HC-41 to HC-44 respectively. Their structural formulas are as follows:

[0155]

[0156] Example 23

[0157] HB (or HC) reacts with p-, m-, and o-aminomethylcyclohexanoic acid respectively. The 2-position substitution products of HB are HB-45 to HB-48 respectively, and the 2-position substitution products of HC are HC-45 to HC-48 respectively. Their structural formulas are as follows:

[0158]

[0159] Example 24

[0160] HB (or HC) reacts with methyl p-, m-, and o-aminomethylcyclohexanoate respectively. The 2-position substitution products of HB are HB-45-CH 3 ~HB-48-CH 3 ,and the 2-position substitution products of HC are HC-45-CH 3~HC-48-CH 3 , and its structural formula is as follows:

[0161]

[0162] Example 25

[0163] HB (or HC) reacts with p-, m-, and o-aminomethylcyclohexanol respectively. The 2-position substitution products of HB are HB-49 to HB-52 respectively, and the 2-position substitution products of HC are HC-49 to HC-52 respectively. Their structural formulas are as follows:

[0164]

[0165] Example 26

[0166] HB (or HC) reacts with amino quaternary ammonium salts with different chain lengths (counterions are bromide ions or iodide ions) respectively. The 2-position substitution products of HB are HB-53 to HB-56 respectively, and the 2-position substitution products of HC are HC-53 to HC-56 respectively. Their structural formulas are as follows:

[0167]

[0168] Example 27

[0169] HB (or HC) reacts with aminoethanol-different chain length quaternary ammonium salts and aminopropanol-different chain length quaternary ammonium salts respectively (counterions are Br - or I - ), the 2-position substitution products of HB are HB-57 to HB-60 respectively, and the 2-position substitution products of HC are HC-57 to HC-60 respectively. Their structural formulas are as follows:

[0170]

[0171] Example 28

[0172] HB (or HC) reacts with amino propionic acid-quaternary ammonium salt and amino butyric acid-quaternary ammonium salt respectively (counterions are Br - or I - ), the 2-position substitution products of HB are HB-61 to HB-64 respectively, and the 2-position substitution products of HC are HC-61 to HC-64 respectively. Their structural formulas are as follows:

[0173]

[0174] Example 29

[0175] HB (or HC) reacts with amino propionic acid-amino quaternary ammonium salt and amino butyric acid-amino quaternary ammonium salt respectively (counterions are Br- or I - Substances), the 2-position substitution products of HB are HB-65 to HB-68 respectively, and the 2-position substitution products of HC are HC-65 to HC-68 respectively. Their structural formulas are shown as follows:

[0176] Example 30

[0177] HB (or HC) reacts with aminopiperidine, morpholine, and aminopiperidine respectively. The 2-position substitution products of HB are HB-69 to HB-72 respectively, and the 2-position substitution products of HC are HC-69 to HC-72 respectively. Their structural formulas are shown as follows:

[0178]

[0179] Example 31

[0180] HB (or HC) reacts with ethylenediamine, methylethylenediamine, and dimethylethylenediamine respectively. The 2-position substitution products of HB are HB-73 to HB-76 respectively, and the 2-position substitution products of HC are HC-73 to HC-76 respectively. Their structural formulas are shown as follows:

[0181]

[0182] Example 32

[0183] HB (or HC) reacts with dimethylethylenediamine, butylethylenediamine, and cyclohexanediamine respectively. The 2-position substitution products of HB are HB-77 to HB-80 respectively, and the 2-position substitution products of HC are HC-77 to HC-80 respectively. Their structural formulas are shown as follows:

[0184]

[0185] Example 33

[0186] HB (or HC) reacts with hydroxyethyl ethylenediamine and hydroxyethyl ethylenediamine - polyethylene glycol respectively. The 2-position substitution products of HB are HB-81, HB-81-PEGn, HB-82, HB-82-PEGn respectively, and the 2-position substitution products of HC are HC-81, HC-81-PEGn, HC-82, HC-82-PEGn (n is an integer between 1 and 100). Their structural formulas are shown as follows:

[0187]

[0188] Example 34

[0189] HB (or HC) reacts with hydroxybutylethylenediamine and hydroxybutylethylenediamine - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 83, HB - 83 - PEGn, HB - 84, HB - 84 - PEGn respectively, and the 2 - position substitution products of HC are HC - 83, HC - 83 - PEGn, HC - 84, HC - 84 - PEGn respectively (n is an integer between 1 and 100). Their structural formulas are as follows:

[0190]

[0191] Example 35:

[0192] HB (or HC) reacts with dihydroxyethylethylenediamine and trihydroxyethylethylenediamine - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 85 to HB - 88 respectively, and the 2 - position substitution products of HC are HC - 85 to HC - 88 respectively. Their structural formulas are as follows:

[0193]

[0194] Example 36

[0195] HB (or HC) reacts with hydroxyethylethylenediamine, hydroxyethylethylenediamine methyl ether, and hydroxyethylethylenediamine - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 89, HB - 89 - CH 3 、HB - 89 - PEGn, and the 2 - position substitution products of HC are HC - 89, HC - 89 - CH 3 、HC - 89 - PEGn (n is an integer between 1 and 100). Their structural formulas are as follows:

[0196]

[0197] Example 37

[0198] HB (or HC) reacts with hydroxypropyl ethylenediamine, hydroxypropyl ethylenediamine methyl ether, and hydroxypropyl ethylenediamine - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 90, HB - 90 - CH 3 、HB - 90 - PEGn, and the 2 - position substitution products of HC are HC - 90, HC - 90 - CH 3 、HC - 90 - PEGn (n is an integer between 1 and 100). Their structural formulas are as follows:

[0199]

[0200] Example 38

[0201] HB (or HC) reacts with hydroxybutylethylenediamine, hydroxybutylethylenediamine methyl ether, and hydroxybutylethylenediamine - polyethylene glycol respectively. The 2 - position substitution products of HB are HB - 91, HB - 91 - CH 3 , HB - 91 - PEGn respectively. The 2 - position substitution products of HC are HC - 91, HC - 91 - CH 3 , HC - 91 - PEGn (n is an integer between 1 and 100), and their structural formulas are as follows:

[0202]

[0203] Example 39

[0204] The general structural formula of the hypocrellin derivatives involved in the present invention is as Figure 1 shown. Hypocrellin B (HB) (or deacetylhypocrellin C, HC) reacts with amino derivatives, mainly generating 2 - position amino - substituted hypocrellin derivatives, as shown in formula (I); hypocrellin reacts with ethylenediamine - type derivatives, and hypocrellin ethylenediamine - substituted derivatives can be obtained, as shown in formulas (II) and (III). Taking the reaction of HB and aminopentanol as an example, the mainly generated 2 - position amino - substituted derivative is HB - 4, and the obtained product continues to react with carboxyl - polyethylene glycol for esterification to obtain HB - 4 - PEGn. Its synthesis method is as Figure 2 shown; HC reacts with methylethylenediamine, mainly generating HC - 74 and HC - 75. Its synthesis method is as Figure 3 shown.

[0205] The derivatives shown in formulas (I), (II), and (III) of the present invention have a maximum absorption wavelength of about 580 - 650 nm, and a molar extinction coefficient of 15000 - 40000 M -1 cm -1 , and have a strong light absorption ability in the phototherapy window; under photosensitizing conditions, they can not only efficiently generate singlet oxygen, but also generate a small amount of superoxide radicals. Such derivatives have good light stability and pH stability. The specific test results are as follows:

[0206] 1) Absorption spectrum

[0207] Figure 4 shows the absorption spectra of the related compounds of the present invention. Figure 4 (a) shows that the commercial porphyrin - type photosensitizer PpIX has multi - band absorption, all of which are narrow absorption bands. The maximum absorption wavelength available for phototherapy is 570 nm, and its molar extinction coefficient is less than 8000 M -1 cm -1 ; the maximum absorption wavelength of the commercial photosensitizer Ce6 is 650 nm, and the molar extinction coefficient is about 15000 M -1 cm -1, which also has narrow absorption in the phototherapy window. Therefore, the light absorption ability of commercial PpIX and Ce6 in the phototherapy window is limited. However, the absorption spectral properties of the derivatives in the present invention are completely different. As Figure 4 (b) shows, the maximum absorption peak of HB is around 470 nm. The derivative HB-6-PEG1 in Example 6 of the present invention has a wide and strong absorption in the phototherapy window. It has a very wide absorption band between 500 and 750 nm, and the maximum absorption peak is around 580 nm, which is about 110 nm red-shifted compared to the maximum absorption peak of HB. The molar extinction coefficient is about 20000 M -1 cm -1 or so, showing a strong red light absorption ability. Similarly, HC-74 is the reaction product of deacetylated hypocrellin and 2-methyl ethylenediamine in Example 31 of the present invention. Its absorption spectrum has a greater degree of red shift and is located in the ideal phototherapy window. It has a very wide absorption band between 500 and 750 nm, and the main absorption peak is at 650 nm, showing a strong red light absorption ability. Other derivatives provided by the present invention also have absorption spectra similar to HB-6-PEG1 and HC-74, with a very wide absorption band between 500 and 750 nm, and the maximum absorption peak is at 580 - 650 nm. Therefore, the hypocrellin derivatives disclosed in the present invention have better light absorption ability in the phototherapy window than commercial photosensitizers PpIX and Ce6, showing more prominent red light absorption ability.

[0208] 2) Reactive oxygen species

[0209] Figure 5 shows the reactive oxygen species of the hypocrellin derivatives HB-45 and HC-45 (Example 23) related to the present invention tested by electron spin resonance (ESR). As Figure 5 shown, both HB-45 and HC-45 can efficiently generate reactive oxygen (ROS). Measured by singlet oxygen and superoxide radical scavengers respectively, such derivatives can efficiently generate photosensitizing active species, mainly generating singlet oxygen and also generating a small amount of superoxide radicals. Both of these two reactive oxygen species are beneficial to photodynamic therapy. The derivatives disclosed in other examples of the present invention also have the ability to efficiently generate singlet oxygen and assist in generating a small amount of superoxide radicals.

[0210] 3) Singlet oxygen efficiency

[0211] The singlet oxygen efficiency of the compounds described in the present invention in solution is detected by the DHPA method. Figure 6(a), 6(b) and 6(c) respectively show the photocatalytic degradation curves of HB-73, HC-73 (Example 31) and HC-80 (Example 32). It can be seen that the photosensitizers HB-73, HC-73, and HC-80 significantly generate singlet oxygen under light illumination, thereby significantly degrading DHPA. By comparing and calculating with the singlet oxygen efficiency curve of the reference rose bengal RB, the singlet oxygen efficiencies of HB-73 and HC-73 are 0.38 and 0.40 respectively, while HC-80 has the highest singlet oxygen efficiency reaching 0.45. The singlet oxygen efficiencies of other derivatives of the present invention are between 0.15 and 0.45, and all have the ability to efficiently generate reactive oxygen species.

[0212] 4) Water solubility

[0213] Most of the derivatives disclosed in the present invention contain hydrophilic groups such as polyethylene glycol, quaternary ammonium salts, sulfonic acids, and carboxylic acids, making the photosensitizer molecules have strong water solubility under physiological conditions. Such photosensitizer molecules can be well dissolved in each milliliter of normal saline or glucose injection solution, so that the photosensitive drug can be well transported in the blood vessels during intravenous injection without causing blood vessel blockage. For example, HB-1-PEG8 (Example 1) contains 8 ethylene glycol units and can dissolve more than 5 mg per milliliter of normal saline; HB-1-PEG16 contains 16 ethylene glycol units and can dissolve more than 10 mg per milliliter of normal saline; HC-3-PEG6 in Example 3 can dissolve more than 5 mg per milliliter of normal saline; HB-6-PEG16 in Example 6 can dissolve more than 15 mg per milliliter of normal saline; HC-8-PEG12 in Example 8 can dissolve more than 10 mg per milliliter of normal saline; these photosensitizer molecules all exhibit excellent water solubility. Other derivatives of the present invention containing strongly water-soluble groups also have good water solubility and biocompatibility, and can dissolve 1-20 mg or more of photosensitive drug molecules per milliliter of normal saline.

[0214] 5) Photostability

[0215] The comparison of the photostability between the derivatives of the present invention and commercial photosensitizers is as Figure 7 shown. It can be seen that when irradiated with a 635 nm laser at a light intensity of 20 mW / cm 2 for 30 min, the absorption spectra of HB-2-PEG4 and HC-2-PEG8 (Example 2) did not show a significant decrease, and the absorption intensity at the maximum wavelength decreased by less than 10%, having good photostability; the absorption intensity at the maximum wavelength of the derivatives HC-73 (Example 31) and HC-80 (Example 32) also decreased by less than 10%, also having good photostability. Under the same conditions, when irradiated with a 635 nm laser at 20 mW / cm 2Under light intensity, after 30 minutes of illumination, the maximum absorptions of the commercial photosensitizers Ce6 and HpD decreased by 30% and 50% respectively. Other derivatives of the present invention also have good photostability, and under the same conditions, the absorption intensity at the maximum wavelength decreased by less than 10% basically. Therefore, the hypocrellin derivatives of the present invention have better photostability than commercial photosensitizers.

[0216] 6) pH stability

[0217] The pH stability of the hypocrellin derivatives of the present invention is as Figure 8 shown. For such derivatives, in the range of pH 6.2 - 8.0, there is no obvious change in the absorption spectrum, indicating that such derivatives have good pH stability under physiological conditions. As shown in the figure, for HB-3-PEG2 and HC-3-PEG8 (Example 3), there is no obvious change in their absorption spectra in the range of pH 6.2 - 8.0; for HC-73 (Example 31) and HB-77 (Example 32), there is no obvious change in their absorption spectra in the range of pH 6.2 - 8.0. They have good pH stability in this range. The reason is that the two phenolic hydroxyl groups of hypocrellin are not easily deprotonated under these conditions. While the commercial hematoporphyrin HpD contains two carboxyl groups, which can be deprotonated in the range of pH 6.2 - 8.0, resulting in an obvious change in the absorption spectrum, thus showing the instability of the HpD photosensitizer. Other derivatives of the present invention also have good pH stability under physiological conditions, and the change in the range of pH 6.2 - 8.0 has little effect on the absorption spectrum of the photosensitive drug. Therefore, the hypocrellin derivatives of the present invention have better pH stability than the commercial porphyrin HpD.

[0218] The structures of some hypocrellin derivatives involved in the present invention are as shown in Examples 1 - 38. For some derivatives of hypocrellin with 2-position amino substitution or ethylenediamine substitution, their maximum absorption wavelengths are around 580 - 630 nm, and the molar extinction coefficients can reach 15000 - 40000 M -1 cm -1 , and they have strong light absorption ability in the phototherapy window; under photosensitive conditions, they can efficiently generate reactive oxygen species such as singlet oxygen, and the singlet oxygen efficiency can reach up to about 40% at most; such derivatives have good photostability and pH stability, and some of their photophysical data are shown in Table 1.

[0219] Table 1: Photophysical data of some hypocrellin derivatives of the present invention

[0220]

[0221] Example 40

[0222] Cultivation of tumor cells: Various cell lines (esophageal cancer cell line AKR, gastric cancer cell line MFC, lung cancer cell line A549, liver cancer cell line HepG2, cholangiocarcinoma cell line MCC, colon cancer cell line HCT116, brain cancer cell line G442, head and neck cancer cell line SCC2, tongue cancer cell line TSCCa, nasal cancer cell line KB, oral cancer cell line CAL27, glioblastoma cell line C6, basal cell carcinoma cell line BCC, squamous skin cancer cell line PECA, melanoma cell line B16, cutaneous T-cell lymphoma cell line HH, prostate cancer cell line LNCaP, bladder cancer cell line MBT-2) were provided by the Cell Center of Peking Union Medical College. The culture conditions for the above cells were as follows: RPMI-1640 medium supplemented with 10% FBS, 1% streptomycin (100 μg / mL) and penicillin (100 μg / mL), and cultured in an incubator at 37°C and 5% CO 2 2. The tumor cells were inoculated into a confocal dish with a glass substrate, and a solution of hypocrellin photosensitizing drug (100 μL) was added to the culture medium, and then incubated in an incubator for 4 h. The cells were carefully washed twice with pre-cooled PBS solution to remove the photosensitizing drug that did not enter the cells, and the cultured tumor cells were used for the MTT assay.

[0223] Example 41

[0224] Cell dark toxicity experiment, taking HB-1-PEG8 (Example 1) as an example: The cultured lung cancer cells (A549 cells) were digested with 0.25% trypsin to prepare a single-cell suspension, which was inoculated into a 96-well culture plate and cultured in an incubator. After the cells adhered to the wall, the supernatant culture medium was discarded, and different concentrations of photosensitizer (such as HB-1-PEG8) were added under light-shielded conditions and incubated for 1 h, and the cell viability was detected by the MTT method. 20 μL of MTT was added to each well, and the cells were cultured for another 4 h. Then the supernatant was discarded, and 150 μL of DMSO was added to each well, and the mixture was shaken with a micro shaker for 10 minutes to fully dissolve the purple crystals. The OD value (570 nm) of each well was measured on an enzyme-linked immunosorbent assay (ELISA) reader and the cell viability was calculated. Cell viability = OD value of the experimental group / OD value of the blank group × 100%. As Figure 10-2 shown, the cell toxicity (dark toxicity) study showed that the cell toxicity of HB-1-PEG8 was relatively low, similar to that of the commercial photosensitizing drug HpD. When incubated with a 20 μM concentration of the photosensitizing drug for half an hour, no obvious death of A549 cells was observed, indicating that such photosensitizing drugs basically had no cell toxicity. The dark toxicity of other derivatives of the present invention was similar to that of HB-1-PEG8, and the results were shown in Figures 10-14 and Tables 2-4. Such derivatives basically had no cell toxicity, and the cell viability was above 90% within a concentration of 20 μM.

[0225] Example 42

[0226] Cell phototoxicity experiment, taking HC-1-PEG8 synthesized in Example 1 as an example:

[0227] The experimental procedure was the same as the dark toxicity experiment. A semiconductor laser with a wavelength of 635 nm (20 mW / cm 2 ) was used for irradiation, and the light beam was uniformly and perpendicularly irradiated onto the 96-well culture plate for 1000 s. As Figure 10-1 shown by the cell phototoxicity experiment, HC-1-PEG8 showed very strong killing ability against A549 cells under red light irradiation. More than 90% of A549 cells could be killed in the concentration range of 50 nM, while only about 20% of A549 cells could be killed by the commercial photosensitizer under the same conditions, indicating that the photodynamic effect of such derivatives was significantly better than that of the commercial photosensitizer HpD. The phototoxicity of other derivatives of the present invention was similar to that of HC-1-PEG8, and the results were shown in Figures 10 - 14. The synthesized derivatives could kill more than 80 - 90% of tumor cells in the concentration range of 50 nM, and the half-lethal concentration IC 50 value was about 20 - 30 nM. Therefore, the hypocrellin derivatives disclosed in the present invention had better photodynamic effects than the commercial photosensitizer HpD.

[0228] Example 43

[0229] Animal experiment: All animal experiment operations complied with the regulations on animal use and feeding stipulated by the Chinese Animal Research Ethics Committee. Female Balb / c nude mice (about 20 g) at 4 - 6 weeks old were used to establish tumor models, and 50 μL of the suspension of various tumor cells (5×10 6 cells) were injected into the posterior side of the right thigh of the mice respectively. When the tumor volume grew to about 200 mm 3 , in vivo fluorescence imaging and photodynamic therapy experiments were carried out.

[0230] Small animal fluorescence imaging experiment: After injecting 40 μL of the physiological saline solution of the photosensitive drug (10 mg / mL) into the tumor-bearing mice via the tail vein according to the mouse dose (10 mg / kg), the tumor fluorescence imaging at 0, 1, 2, 4, 6, and 8 hours was collected using small animal in vivo imaging respectively. After 8 h, the tumor-bearing mice were sacrificed by cervical dislocation, and the tumors and main organs were taken out for in vitro fluorescence imaging, and the average fluorescence intensity in the ROS region was statistically analyzed for semi-quantitative analysis.

[0231] Photodynamic therapy experiment: When the tumor volume grew to 200 mm 3 , the tumor-bearing mice were randomly divided into 3 groups (5 mice in each group): without any treatment (Group I); only injecting the photosensitive drug via the tail vein (100 μL, 500 μM) (Group II); injecting the photosensitive drug via the tail vein (100 μL, 500 μM) and irradiating the tumor site with a 635 nm laser (10 min, 0.1 W cm -2)(Group III). The tumor volume and body weight were measured every other day after laser treatment, and the survival rate was recorded.

[0232] Example 44

[0233] As described above, the drug activities of the hypocrellin derivatives of the present invention were tested, and the specific results are as follows:

[0234] 1) Confocal fluorescence imaging of hypocrellin derivatives in cancer cells

[0235] The cellular fluorescence imaging of the hypocrellin derivatives of the present invention is as Figure 9 shown. The photosensitizer HB-1-PEG12 has good water solubility and biocompatibility. When co-incubated with lung cancer cells (A549), it was found that the photosensitizer could rapidly enter the lysosomes of the cells and produce red fluorescence in the cells, indicating that HB-1-PEG12 can be used for fluorescence imaging of lung cancer cells to track the enrichment, distribution, and metabolism of the drug in vivo through fluorescence detection. DCFH-DA was used to detect singlet oxygen in the cells. The photosensitizer HB-1-PEG12 and the fluorescent probe DCFH-DA were co-incubated in the cells. As the irradiation time increased to 120 s, the green fluorescence gradually increased, indicating an increase in singlet oxygen in the cells. Other derivatives of the present invention can also well enter the lysosomes of lung cancer cells and perform cellular fluorescence imaging to track the enrichment, distribution, and metabolism of the drug in vivo through fluorescence detection.

[0236] 2) Dark toxicity and phototoxicity tests of hypocrellin derivatives on digestive tract tumor cells

[0237] Digestive tract tumor cells mainly include esophageal cancer cells, gastric cancer cells, lung cancer cells, liver cancer cells, cholangiocarcinoma cells, and colon cancer cells.

[0238] HB-1-PEG4 and HC-1-PEG8 (Example 1) were co-incubated with esophageal cancer AKR cells. As Figure 10-1 (a) shows, the study on the cytotoxicity (dark toxicity) of the photosensitizing drug without light irradiation indicates that whether it has 4 or 8 PEG units, the cytotoxicity of HB-1-PEG4 and HC-1-PEG8 is relatively small, similar to that of the commercial photosensitizing drug HpD. Incubating with a 20 μM concentration of the photosensitizer HB-1-PEG4 or HC-1-PEG8 for half an hour, no obvious death of esophageal cancer cells was observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 10-1 (b) shows, the study on the cell phototoxicity indicates that the photosensitizing drug shows very strong lethality to esophageal cancer cells under red light irradiation at 635 nm (10 min, 0.1 W cm -2 ). A 50 nM concentration of HB-1-PEG4 can kill more than 90% of esophageal cancer cells, and the half-lethal concentration IC50 The value (the drug concentration required to kill half of the tumor cells) is approximately 25 nM; HC-1-PEG8 at a concentration of 50 nM can kill more than 90% of esophageal cancer cells, and the half-lethal concentration IC 50 value is approximately 25 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 30% of esophageal cancer cells, indicating that the photodynamic killing effect of such derivatives on esophageal cancer is significantly better than that of commercial photosensitizers.

[0239] The derivatives in the present invention were incubated with lung cancer cell line A549. As Figure 10-2 (a) shows, the dark toxicity study of the cells indicated that HB-3-PEG8 and HC-3-PEG16 (Example 3) had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. Lung cancer cells were incubated with a photosensitizer at a concentration of 20 μM of HB-3-PEG8 or HC-3-PEG16 for half an hour, and no obvious death of lung cancer cells was observed, indicating that such photosensitizers have no cytotoxicity. As Figure 10-2 (b) shows, the cell phototoxicity experiment indicated that the photosensitizing drug showed very strong killing power to lung cancer cells under red light irradiation. HB-3-PEG8 or HC-3-PEG16 at a concentration of 50 nM could kill more than 90% of lung cancer cells, and the half-lethal concentration IC 50 value was approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD could only kill about 20% of lung cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives was significantly better than that of commercial photosensitizers.

[0240] The derivatives in the present invention were incubated with liver cancer cell line HepG2. As Figure 10-3 (a) shows, the dark toxicity study of the photosensitizing drug indicated that both HB-6-PEG2 and HC-6-PEG6 prepared in Example 6 had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug HpD. Liver cancer cells were incubated with a photosensitizer at a concentration of 20 μM for half an hour, and no obvious death of liver cancer cells was observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 10-3 (b) shows, the cell phototoxicity study indicated that the photosensitizing drug showed very strong killing power to liver cancer cells under red light irradiation. HB-6-PEG2 or HC-6-PEG12 at a concentration of 50 nM could kill more than 85% of liver cancer cells, and the half-lethal concentration IC 50 value was approximately 30 nM; while under the same conditions, the commercial hematoporphyrin derivative HpD could only kill about 20% of liver cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives was significantly better than that of commercial photosensitizers.

[0241] The derivatives in the present invention were incubated with colon cancer cell line HCT116. As Figure 10-4As shown in (a), the dark toxicity study of the photosensitizing drugs indicates that both HB-14-PEG6 and HC-14-PEG12 prepared in Example 13 have relatively low cytotoxicity, similar to that of the commercial photosensitizing drug HpD. When colon cancer cells were incubated with a photosensitizer at a concentration of 20 μM for half an hour, no obvious death of colon cancer cells was observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 10-4 As shown in (b), the cell phototoxicity study indicates that the photosensitizing drugs show very strong killing power against colon cancer cells under red light irradiation. Both HB-14-PEG6 and HC-14-PEG12 at a concentration of 50 nM can kill more than 85% of colon cancer cells, and the half-lethal concentration IC 50 value is approximately 25 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of colon cancer cells, indicating that the photodynamic effect of such derivatives is significantly better than that of the commercial photosensitizer HpD.

[0242] When the derivatives in the present invention were incubated with cholangiocarcinoma cells MCC, as Figure 10-5 As shown in (a), the dark toxicity study of the photosensitizing drugs indicates that both HB-19-PEG4 and HC-19-PEG8 prepared in Example 15 have relatively low cytotoxicity, similar to that of the commercial photosensitizing drug HpD. When cholangiocarcinoma cells were incubated with a photosensitizer at a concentration of 20 μM for half an hour, no obvious death of cholangiocarcinoma cells was observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 10-5 As shown in (b), the cell phototoxicity study experiment indicates that the photosensitizing drugs show very strong killing power against cholangiocarcinoma cells under red light irradiation. Both HB-19-PEG4 and HC-19-PEG8 at a concentration of 50 nM can kill more than 85% of cholangiocarcinoma cells, and the half-lethal concentration IC 50 value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of cholangiocarcinoma cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of the commercial photosensitizer HpD.

[0243] When the derivatives in the present invention were incubated with gastric cancer cells MFC, as Figure 10-6 As shown in (a), the dark toxicity study of the photosensitizing drugs indicates that both HB-45 and HC-45 prepared in Example 23 have relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. When gastric cancer cells were incubated with a photosensitizer at a concentration of 20 μM for half an hour, no obvious death of MFC cells was observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 10-6 As shown in (b), the cell phototoxicity study indicates that the photosensitizing drugs show very strong killing power against gastric cancer cells under red light irradiation. Both HB-45 and HC-45 at a concentration of 50 nM can kill more than 85% of gastric cancer cells, and the IC 50The value is about 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of gastric cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of commercial photosensitizers.

[0244] The above details illustrate that some of the hypocrellin derivatives disclosed in the present invention can efficiently kill digestive tract tumor cells such as esophageal cancer AKR, gastric cancer MFC, lung cancer A549, liver cancer HepG2, cholangiocarcinoma MCC, and colon cancer HCT116. Whether it is a derivative of hypocrellin (HB) or deacetylated hypocrellin (HC), there is basically no damage to cells without light irradiation, while it has a strong ability to inactivate the above-mentioned digestive tract tumor cells under light irradiation. In addition, hypocrellin derivatives with different PEG chain lengths, different sulfonic acid chain lengths, and different quaternary ammonium salt chain lengths all show good photodynamic cell-inactivating ability against tumor cells. Therefore, such derivatives can all efficiently kill digestive tract tumor cells such as esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, and colon cancer, and the relevant data are shown in Table 2.

[0245] Table 2: MTT data of some hypocrellin derivatives of the present invention against digestive tract tumor cells

[0246]

[0247] 3) Dark toxicity and phototoxicity tests of hypocrellin derivatives on head and neck tumor cells

[0248] Head and neck tumor cells mainly include head and neck cancer cells, brain cancer cells, tongue cancer cells, nasal cancer cells, oral cancer cells, and glioma cells.

[0249] The hypocrellin derivatives disclosed in the present invention can efficiently kill head and neck tumor cells such as brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer, and glioma under 635 nm laser irradiation. Taking the derivatives of hypocrellin disclosed in the examples as an example, the phototoxicity and dark toxicity effects on head and neck tumor cells are described.

[0250] The derivatives in the present invention were incubated with brain cancer cells G442. The study on the dark toxicity of the photosensitizing drug showed that whether it is linked with 4 PEG or 8 PEG units, the cytotoxicity of HB-10-PEG4 and HC-10-PEG8 (Example 10) is relatively small, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. Brain cancer cells were incubated with 20 μM concentration of the photosensitizers HB-10-PEG4 and HC-10-PEG8 for half an hour, and no obvious death of brain cancer cells was observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 11(a) The cell phototoxicity study shows that under red light irradiation, the photosensitizing drugs exhibit very strong killing power against brain cancer cells. Both HB-10-PEG4 and HC-10-PEG8 at a concentration of 50 nM can kill more than 85% of brain cancer cells, and the half-lethal concentration IC 50 value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of brain cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of the commercial photosensitizer HpD.

[0251] When the derivatives in the present invention are incubated with head and neck cancer cells SCC2, the dark toxicity study of the photosensitizing drugs shows that both HB-12-PEG8 and HC-12-PEG12 prepared in Example 12 have relatively low cytotoxicity, similar to that of the commercial photosensitizing drug HpD. When head and neck cancer cells are incubated with a photosensitizer at a concentration of 20 μM for half an hour, no obvious death of head and neck cancer cells is observed, indicating that such photosensitizers have no cytotoxicity. As Figure 11 (b) The cell phototoxicity study shows that under red light irradiation, the photosensitizing drugs exhibit very strong killing power against head and neck cancer cells. Both HB-12-PEG8 and HC-12-PEG12 at a concentration of 50 nM can kill more than 85% of head and neck cancer cells, and the half-lethal concentration IC 50 value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of head and neck cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of the commercial photosensitizer HpD.

[0252] When the derivatives in the present invention are incubated with tongue cancer cells TSCCa, the dark toxicity study of the photosensitizing drugs shows that both HB-20-PEG6 and HC-20-PEG12 prepared in Example 16 have relatively low cytotoxicity, similar to that of the commercial photosensitizing drug HpD. When tongue cancer cells are incubated with a photosensitizer at a concentration of 20 μM for half an hour, no obvious death of tongue cancer cells is observed, indicating that such photosensitizers basically have no cytotoxicity. As Figure 11 (c) The cell phototoxicity study shows that under red light irradiation, the photosensitizing drugs exhibit very strong killing power against tongue cancer cells. Both HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM can kill more than 85% of tongue cancer cells, and the half-lethal concentration IC 50 value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of tongue cancer cells, indicating that the photodynamic effect of such derivatives is significantly better than that of the commercial photosensitizer.

[0253] When the derivatives in the present invention are incubated with nasal cancer cells KB, the dark toxicity study of the photosensitizing drugs shows that both HB-29 and HC-29 prepared in Example 19 have relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. As Figure 12(a) The cell phototoxicity study experiment shows that the photosensitizing drug exhibits very strong killing power against nasal cancer cells under red light irradiation. Both HB-29 and HC-29 at a concentration of 50 nM can kill more than 90% of KB cells, and the half-lethal concentration IC 50 value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of KB cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of the commercial photosensitizer HpD.

[0254] When the derivatives in the present invention are incubated with oral cancer cells CAL27, the dark toxicity study of the photosensitizing drug shows that both HB-61 and HC-61 prepared in Example 28 have relatively low cytotoxicity, similar to the commercial photosensitizing drug hematoporphyrin HpD. As Figure 12 (b) The cell phototoxicity study shows that the photosensitizing drug exhibits very strong killing power against oral cancer cells under red light irradiation. Both HB-61 and HC-61 at a concentration of 50 nM can kill more than 90% of oral cancer cells, and the half-lethal concentration IC 50 value is approximately 25 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 25% of oral cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of the commercial photosensitizer HpD.

[0255] When the derivatives in the present invention are incubated with glioma C6 cells, the dark toxicity study of the photosensitizing drug shows that the hypocrellin derivatives HB-73 prepared in Example 31 and HC-80 prepared in Example 32 have relatively low cytotoxicity against glioma, similar to the commercial photosensitizing drug hematoporphyrin HpD. As Figure 12 (c) The cell phototoxicity study shows that the photosensitizing drug exhibits very strong killing power against glioma cells under red light irradiation. HB-73 and HC-80 at a concentration of 50 nM can kill more than 85% of glioma cells, and the half-lethal concentration IC 50 value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of C6 cells, indicating that the photodynamic effect of such hypocrellin derivatives is significantly better than that of the commercial photosensitizer HpD.

[0256] The above has detailedly illustrated with examples that some of the hypocrellin derivatives disclosed in the present invention can efficiently kill head and neck facial tumor cells such as brain cancer G442, head and neck cancer SCC2, tongue cancer TSCCa, nasal cancer KB, oral cancer CAL27, and glioma C6. Whether it is a derivative of HB or HC, without light irradiation, there is basically no damage to the cells, while under light irradiation, it has a strong ability to inactivate the above-mentioned digestive tract tumor cells. In addition, hypocrellin derivatives with different lengths of PEG chains, different lengths of sulfonic acid chains, and different lengths of quaternary ammonium salt chains all show good ability to photodynamically inactivate tumor cells. Therefore, such derivatives can all efficiently kill head and neck facial tumor cells such as brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer, and glioma. The relevant data are shown in Table 3.

[0257] Table 3: MTT data of some hypocrellin derivatives of the present invention on head and neck facial tumor cells

[0258]

[0259] 4) Dark toxicity and phototoxicity tests of hypocrellin derivatives on reproductive and urinary system tumor cells

[0260] The hypocrellin derivatives disclosed in the present invention can efficiently kill reproductive and urinary system tumor cells such as basal cell carcinoma, cutaneous T-cell lymphoma, melanoma, squamous cell carcinoma of the skin, prostate cancer, and bladder cancer under 635 nm laser irradiation. Taking the hypocrellin derivatives disclosed in the examples as an example, the phototoxicity and dark toxicity effects on skin tumors and urinary system tumor cells are described.

[0261] The derivatives in the present invention were incubated with basal cell carcinoma cells BCC. The study on the dark toxicity of the photosensitizing drug showed that the cytotoxicities of HB-4-PEG8 and HC-4-PEG16 (Example 4) were both small, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. Incubating with a photosensitizer at a concentration of 20 μM for half an hour, no obvious death of basal cell carcinoma cells was seen, indicating that such photosensitizers basically had no cytotoxicity. As Figure 13 (a) shows, the study on the phototoxicity of the photosensitizing drug showed that under red light irradiation, the photosensitizing drug showed very strong killing power on basal cell carcinoma cells. HB-4-PEG8 and HC-4-PEG16 at a concentration of 50 nM could both kill more than 90% of basal cell carcinoma cells, and the half-lethal concentration IC 50 value was about 30 nM; while under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD could only kill about 20% of basal cell carcinoma cells, indicating that the photodynamic effect of such hypocrellin polyethylene glycol derivatives was significantly better than that of the commercial photosensitizer hematoporphyrin HpD.

[0262] The derivatives in the present invention were incubated with melanoma cells B16. The study on the dark toxicity of the photosensitizing drugs showed that both HB-9-PEG4 and HC-9-PEG8 prepared in Example 9 had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. As Figure 13 (b) shows, the study on the phototoxicity of the photosensitizing drugs indicated that under red light irradiation, the photosensitizing drugs showed very strong killing power against melanoma cells. HB-9-PEG4 and HC-9-PEG8 at a concentration of 50 nM could both kill more than 85% of melanoma cells, and the half-lethal concentration IC 50 value was approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD could only kill about 20% of melanoma cells, indicating that the photodynamic effect of such hypocrellin derivatives was significantly better than that of the commercial photosensitizer HpD.

[0263] The derivatives in the present invention were incubated with squamous skin cancer cells PECA. The study on the dark toxicity of the photosensitizing drugs showed that both HB-36 and HC-36 prepared in Example 20 had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug HpD. As Figure 13 (c) shows, the study on the phototoxicity of the photosensitizing drugs indicated that under red light irradiation, the photosensitizing drugs showed very strong killing power against PECA cells. HB-36 and HC-36 at a concentration of 50 nM could kill more than 85% of skin cancer cells, and the half-lethal concentration IC 50 value was approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD could only kill about 20% of PECA cells, indicating that the photodynamic effect of such hypocrellin derivatives was significantly better than that of the commercial photosensitizer HpD.

[0264] The derivatives in the present invention were incubated with prostate cancer cells LNCaP. The study on the dark toxicity of the photosensitizing drugs showed that both HB-70 and HC-71 prepared in Example 30 had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. As Figure 14 (a) shows, the study on the phototoxicity of the photosensitizing drugs indicated that under red light irradiation, the photosensitizing drugs showed very strong killing power against prostate cancer cells. HB-70 and HC-71 at a concentration of 50 nM could both kill more than 85% of prostate cancer cells, and the half-lethal concentration IC 50 value was approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD could only kill about 20% of prostate cancer cells, indicating that the photodynamic effect of such hypocrellin derivatives was significantly better than that of the commercial photosensitizer HpD.

[0265] The derivatives in the present invention were incubated with bladder cancer cells MBT-2. The study on the dark toxicity of the photosensitizing drugs showed that both HB-73 and HC-73 prepared in Example 31 had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. As Figure 14(b) The study on the phototoxicity of cells showed that under red light irradiation, the photosensitizing drugs exhibited very strong killing effects on bladder cancer cells. Both HB-73 and HC-73 at a concentration of 50 nM could kill more than 85% of bladder cancer cells, and the half-lethal concentration IC 50 value was approximately 25 nM; while under the same conditions, the commercial photosensitizer HpD could only kill about 20% of bladder cancer cells, indicating that the photodynamic effects of such hypocrellin polyethylene glycol derivatives were significantly superior to those of commercial photosensitizers.

[0266] When the derivatives in the present invention were incubated with skin T cell lymphoma cells HH, the study on the dark toxicity of the photosensitizing drugs showed that both HC-80 prepared in Example 32 and HC-81 prepared in Example 33 had relatively low cytotoxicity, similar to that of the commercial photosensitizing drug hematoporphyrin HpD. As Figure 14 (c) The study on the phototoxicity of cells showed that under red light irradiation, the photosensitizing drugs exhibited very strong killing effects on skin T cell lymphoma cells. Both HC-80 and HC-81 at a concentration of 50 nM could kill more than 85% of HH cells, and the half-lethal concentration IC 50 value was approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD could only kill 20% of HH cells, indicating that the photodynamic effects of such hypocrellin derivatives were significantly superior to those of commercial photosensitizers.

[0267] The above details illustrate by examples that some of the hypocrellin derivatives disclosed in the present invention can efficiently kill skin and urinary system tumor cells such as basal cell carcinoma BCC, squamous cell carcinoma of the skin PECA, melanoma B16, skin T cell lymphoma HH, prostate cancer LNCaP, and bladder cancer MBT-2. Whether it is a derivative of hypocrellin or deacetyhypocrellin, without light irradiation, there is basically no damage to cells, while under light irradiation, it has a strong ability to inactivate the above-mentioned digestive tract tumor cells. In addition, hypocrellin derivatives with different PEG chain lengths, different sulfonic acid chain lengths, and different quaternary ammonium salt chain lengths all exhibit good photodynamic cell-inactivating ability against tumor cells. Therefore, such derivatives can all efficiently kill reproductive and urinary system tumor cells such as basal cell carcinoma, squamous cell carcinoma of the skin, melanoma, skin T cell lymphoma, prostate cancer, and bladder cancer, and the detailed data are shown in Table 4.

[0268] Table 4: MTT data of some hypocrellin derivatives of the present invention on skin tumors and reproductive and urinary tumors

[0269]

[0270] The above research shows that: by changing the molecular structure, as well as different PEG chain lengths and end groups, the hypocrellin derivatives of the present invention have good photodynamic killing effects on various tumor cells. Figures 10-14 and Tables 2-4 illustrate that such derivatives have almost no cytotoxicity without light irradiation, and have obvious photodynamic effects under light irradiation. The photosensitizing drug at a concentration of 50 nM can efficiently kill most various tumor cells, and the half-lethal concentration IC 50 value is about 20-30 nM, which is 1-2 orders of magnitude lower than the IC 50 value of the commercial photosensitizer HpD under the same conditions. Other derivatives of the present invention also have the dark toxicity and phototoxicity as shown in Figures 10-14. Therefore, the hypocrellin derivatives substituted with amino groups or ethylenediamine at the 2-position disclosed in the present invention have better dark toxicity and photodynamic effects than the commercial photosensitizer HpD.

[0271] Example 45 Animal Imaging

[0272] The fluorescence imaging of the aforementioned cells ( Figure 9 ) shows that the derivatives provided by the present invention can enter tumor cells well and produce near-infrared fluorescence imaging. In order to study the enrichment and metabolism processes of such derivatives at the tumor in vivo, this application uses tumor-bearing mice with various different tumor cells as models, and studies the enrichment process of such derivatives at the tumors of mice through small animal in vivo fluorescence imaging. Derivatives with good water solubility such as those containing polyethylene glycol, sulfonic acid group, carboxylic acid group, quaternary ammonium salt, etc. achieve targeted enrichment of the drug at the tumor through tail vein injection; while for derivatives with slightly worse water solubility, the photosensitizing drug is enriched in the tumor area through intratumoral injection of the tumor-bearing mice.

[0273] 1) Drug Enrichment in Digestive Tract Tumor Mice (Esophageal Cancer, Gastric Cancer, Lung Cancer, Liver Cancer, Cholangiocarcinoma, Colon Cancer)

[0274] Subcutaneous tumors were inoculated with esophageal cancer AKR cells to obtain a subcutaneous mouse tumor model of esophageal cancer. HB-1-PEG4 (Example 1) was used as the photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg, and its fluorescence imaging behavior in the esophageal cancer-bearing mice was observed. The fluorescence signals at the tumor sites at 4 hours were collected using a multi-spectral small animal in vivo imaging system. As Figure 15 (a) shows, after several cycles in vivo through intravenous injection, the drug molecule HB-1-PEG4 has a certain drug enrichment at the tumor after about 4 hours through passive targeting, which belongs to medium-level enrichment. The reason is that this derivative molecule only contains 4 polyethylene glycol units, and the water solubility of the whole drug molecule is not enough, resulting in medium-level enrichment at the tumor.

[0275] Subcutaneous tumors were inoculated with gastric cancer MFC cells to obtain a subcutaneous mouse tumor model of gastric cancer. HC-3-PEG12 (Example 3) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 15 (b) shows, after several cycles in vivo by intravenous injection, the drug molecule HC-3-PEG12 has strong drug accumulation at the tumor after about 4 hours through passive targeting. The reason is that this derivative molecule contains 12 polyethylene glycol units, making the water solubility of the drug molecule better, and the hydrophilic-hydrophobic ratio of the whole molecule appropriate, resulting in strong accumulation at the tumor.

[0276] Subcutaneous tumors were inoculated with lung cancer A549 cells to obtain a subcutaneous mouse tumor model of lung cancer. HB-6-PEG8 (Example 6) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior in the lung cancer-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 15 (c) shows, after several cycles in vivo by intravenous injection, the drug molecule HB-6-PEG8 has strong accumulation at the tumor after about 4 hours through passive targeting. The reason is that this derivative molecule contains 8 polyethylene glycol units, making the water solubility of the molecule better, and the hydrophilic-hydrophobic ratio of the whole molecule appropriate, resulting in strong accumulation at the tumor.

[0277] Subcutaneous tumors were inoculated with liver cancer HepG2 cells to obtain a subcutaneous mouse tumor model of liver cancer. HB-7-PEG16 (Example 7) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior in the liver cancer-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 15 (d) shows, after several cycles in vivo by intravenous injection, HB-7-PEG16 has strong accumulation at the tumor after about 4 hours through passive targeting. The reason is that this molecule contains 16 polyethylene glycol units, making the water solubility of the whole drug molecule better and the hydrophilic-hydrophobic ratio appropriate, resulting in strong accumulation at the tumor.

[0278] Subcutaneous tumors were inoculated with cholangiocarcinoma MCC cells to obtain a subcutaneous mouse tumor model of cholangiocarcinoma. HC-9-PEG10 (Example 9) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior in the cholangiocarcinoma-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 15(e), After several cycles in vivo by intravenous injection, HC-9-PEG10 has a strong enrichment at the tumor site after about 4 h through passive targeting. The reason is that this molecule contains 10 polyethylene glycol units, making the whole drug molecule have good water solubility and a suitable hydrophilic-hydrophobic ratio, thus resulting in a strong enrichment at the tumor site.

[0279] Subcutaneous tumors were inoculated with colon cancer HCT116 cells to obtain a subcutaneous mouse tumor model of colon cancer. HC-1-PEG50 (Example 1) was used as a photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the colon cancer-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 15 (f) shows, After several cycles in vivo by intravenous injection, HC-1-PEG50 has a strong enrichment at the tumor site after about 4 h through passive targeting. The reason is that this derivative contains 50 polyethylene glycol units, making the whole drug molecule have good water solubility and a suitable hydrophilic-hydrophobic ratio, thus resulting in a strong enrichment at the tumor site.

[0280] 2) Drug enrichment in mice with head and neck tumors (brain cancer, head and neck cancer, tongue cancer, oral cancer, nasal cancer, glioma)

[0281] Subcutaneous tumors were inoculated with brain cancer G442 cells to obtain a subcutaneous mouse tumor model of brain cancer. HB-11-PEG6 (Example 11) was used as a photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the brain cancer-bearing mice was observed, and the fluorescence imaging at the tumor site was collected at 4 h. As Figure 16 (a) shows, After several cycles in vivo by intravenous injection, HB-11-PEG6 has a certain enrichment ability at the tumor site after about 4 h through passive targeting, belonging to moderate enrichment. The reason is that this derivative molecule only contains 6 polyethylene glycol units, making the water solubility of the whole drug molecule insufficient, thus resulting in a moderate enrichment at the tumor site.

[0282] Subcutaneous tumors were inoculated with head and neck cancer SCC2 cells to obtain a subcutaneous mouse model of head and neck cancer. HC-18-PEG8 (Example 15) was used as a photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the head and neck cancer-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 16As shown in (b), after several cycles in vivo by intravenous injection, HC-18-PEG8 has a strong enrichment at the tumor site after about 4 h through passive targeting. The reason is that this derivative contains 8 polyethylene glycol units, making the whole drug molecule have good water solubility and an appropriate hydrophilic-hydrophobic ratio, thus resulting in a strong enrichment at the tumor site.

[0283] Subcutaneous tumors were inoculated with tongue cancer TSCCa cells to obtain a subcutaneous mouse tumor model of tongue cancer. HC-45 (Example 23) was used as a photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the tongue cancer-bearing mice was observed, and the fluorescence signals at the tumor site were collected at 4 h. As Figure 16 As shown in (c), after several cycles in vivo by intravenous injection, HC-45 has a medium-strength enrichment at the tumor site after about 4 h through passive targeting. The reason is that this derivative molecule does not contain polyethylene glycol units and only contains one carboxyl group, and its water solubility is also insufficient, thus resulting in a medium-strength enrichment at the tumor site.

[0284] Subcutaneous tumors were inoculated with oral cancer CAL27 cells to obtain a subcutaneous mouse tumor model of oral cancer. HC-57 (Example 27) was used as a photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the oral cancer-bearing mice was observed, and the fluorescence signals at the tumor site were collected at 4 h. As Figure 16 As shown in (d), after several cycles in vivo by intravenous injection, HC-57 has a strong enrichment ability at the tumor site after about 4 h through passive targeting. The reason is that although this derivative does not contain polyethylene glycol units, it contains water-soluble groups such as carboxyl groups and quaternary ammonium salts, making the whole drug molecule have good water solubility and an appropriate hydrophilic-hydrophobic ratio of the whole molecule, thus resulting in a strong enrichment at the tumor site.

[0285] Subcutaneous tumors were inoculated with nasal cancer KB cells to obtain a subcutaneous mouse tumor model of nasal cancer. HB-65 (Example 29) was used as a photosensitizing drug and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the nasal cancer-bearing mice was observed, and the fluorescence signals at the tumor site were collected at 4 h. As Figure 16 As shown in (e), after several cycles in vivo by intravenous injection, HB-65 has a strong enrichment at the tumor site after about 4 h through passive targeting. The reason is that this derivative contains hydrophilic groups such as quaternary ammonium salts and amide groups, and the whole molecule has good water solubility and an appropriate hydrophilic-hydrophobic ratio, thus resulting in a strong enrichment at the tumor site.

[0286] The subcutaneous glioma mouse tumor model was established by inoculating subcutaneous tumors with glioma C6 cells. HC-81-PEG16 (Example 33) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the glioma-bearing mice was observed, and the fluorescence signals at the tumor sites were collected at 4 hours. As Figure 16 (f) shows, after several cycles in vivo by intravenous injection, HC-81-PEG16 has strong enrichment at the tumor site around 4 h through passive targeting. The reason is that this derivative contains 16 polyethylene glycol units, making the whole drug molecule have good water solubility and an appropriate hydrophilic-hydrophobic ratio, resulting in strong enrichment at the tumor site.

[0287] 3) Drug enrichment in mice with skin and urinary system tumors (basal cell carcinoma, squamous cell carcinoma of the skin, melanoma, cutaneous T-cell lymphoma, prostate cancer, bladder cancer)

[0288] The subcutaneous basal cell carcinoma mouse tumor model was established by inoculating subcutaneous tumors with basal cell carcinoma BCC cells. HB-5-PEG10 (Example 5) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the basal cell carcinoma-bearing mice was observed, and the fluorescence signals at the tumor sites were collected at 4 hours. As Figure 17 (a) shows, after several cycles in vivo by intravenous injection, HB-5-PEG10 has strong enrichment at the tumor site around 4 h through passive targeting. The reason is that this derivative contains 10 polyethylene glycol units, making the whole drug molecule have good water solubility and an appropriate hydrophilic-hydrophobic ratio, resulting in strong enrichment at the tumor site.

[0289] The subcutaneous squamous cell carcinoma mouse model was established by inoculating subcutaneous tumors with squamous cell carcinoma PECA cells. HC-8-PEG12 (Example 8) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the squamous cell carcinoma-bearing mice was observed, and the fluorescence signals at the tumor sites were collected at 4 hours. As Figure 17 (b) shows, after several cycles in vivo by intravenous injection, HC-8-PEG12 has strong enrichment at the tumor site around 4 h through passive targeting. The reason is that this molecule contains 8 polyethylene glycol units, and the whole drug molecule has good water solubility and an appropriate hydrophilic-hydrophobic ratio, resulting in strong enrichment at the tumor site.

[0290] A subcutaneous tumor was inoculated with melanoma B16 cells to obtain a subcutaneous mouse tumor model of melanoma. HB-64 (Example 28) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in melanoma-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 17 (c) shows, after several cycles in vivo by intravenous injection, HB-64 has strong enrichment at the tumor site around 4 h through passive targeting. The reason is that although this molecule does not contain a polyethylene glycol unit, it contains water-soluble groups such as carboxylate and quaternary ammonium salts, making the water solubility of the whole drug molecule good and the hydrophilic-hydrophobic ratio appropriate, thus resulting in strong enrichment at the tumor site.

[0291] A subcutaneous tumor was inoculated with cutaneous T-cell lymphoma HH cells to obtain a subcutaneous mouse tumor model of lymphoma. HB-89-PEG16 (Example 36) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in cutaneous T-cell lymphoma-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 17 (d) shows, after several cycles in vivo by intravenous injection, HB-89-PEG16 has strong enrichment at the tumor site around 4 h through passive targeting. The reason is that this derivative contains 16 polyethylene glycol units, making the water solubility of the whole drug molecule good and the hydrophilic-hydrophobic ratio appropriate, thus resulting in strong enrichment at the tumor site.

[0292] A subcutaneous tumor was inoculated with prostate cancer LNCaP cells to obtain a subcutaneous mouse tumor model of prostate cancer. HC-90-PEG30 (Example 37) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in prostate cancer-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As Figure 17 (e) shows, after several cycles in vivo by intravenous injection, HC-90-PEG30 has strong enrichment at the tumor site around 4 h through passive targeting. The reason is that this molecule contains 30 polyethylene glycol units, and the water solubility of the whole drug molecule is good and the hydrophilic-hydrophobic ratio is appropriate, thus resulting in strong enrichment at the tumor site.

[0293] A subcutaneous tumor was inoculated with bladder cancer MBT-2 cells to obtain a subcutaneous mouse tumor model of bladder cancer. HC-91-PEG16 (Example 38) was used as a photosensitizing drug and injected into the tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in bladder cancer-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. Its animal fluorescence imaging is as Figure 17(f). It can be seen that after several cycles in vivo through intravenous injection, the drug molecule HC-91-PEG16 has a strong enrichment ability at the tumor site after about 4 h through passive targeting. The reason is that this derivative molecule contains 16 polyethylene glycol units, making the water solubility of the whole drug molecule better and the hydrophilic-hydrophobic ratio of the whole molecule appropriate, thus resulting in a strong enrichment at the tumor site.

[0294] The above Figures 15-17 results show that the hypocrellin derivative provided in the present invention can be administered to tumor-bearing mice by the method of tail vein injection. Through small animal in vivo imaging, it can be seen that these drugs produce ultra-strong or moderate fluorescence imaging at the tumor sites of mice, indicating that such derivatives can be used as photosensitizing drugs for fluorescence imaging at the tumor sites of tumor-bearing mice and can be used for the resection of tumor boundaries under the guidance of fluorescence-mediated materials during surgery. The tumors include: esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell skin cancer, melanoma, cutaneous T-cell lymphoma, prostate cancer, bladder cancer.

[0295] For the hypocrellin derivatives with slightly poor water solubility in the present invention, they can be administered by intratumoral injection. Dissolve the hypocrellin derivative in DMSO (dimethyl sulfoxide) and dilute it 5-10 times with physiological saline for intratumoral injection. Taking the subcutaneous mouse tumor model of basal cell carcinoma (BCC cells) as an example, the derivative (Example 19 of HB-28) is used as a photosensitizing drug and injected into tumor-bearing mice by intratumoral injection at a dose of 10 mg / kg, and the fluorescence imaging behavior in the tumor-bearing mice is observed. The fluorescence signals at the tumor sites are collected respectively using a small animal imaging system for 4 hours. As Figure 18 (a) shows, the drug molecule HB-28 is injected into tumor cells and tissues by intratumoral injection, and it has a strong enrichment ability at the tumor site.

[0296] The present invention also discloses the enrichment of other derivatives with poor water solubility through intratumoral administration in different tumor cells. By a similar administration method, the fluorescence imaging behavior in tumor-bearing mice in tumor cells was observed, and the fluorescence signals at the tumor site at 4 hours were collected using a multispectral small animal in vivo imaging system. For example, the fluorescence imaging of the photosensitizing drug HC-36 (Example 20) in mice with squamous skin cancer PECA cells as shown in 18(b); the fluorescence imaging of the photosensitizing drug HB-45 (Example 23) in mice with melanoma B16 cells as shown in 18(c); the fluorescence imaging of the photosensitizing drug HC-73 (Example 31) in mice with cutaneous T-cell lymphoma HH cells as shown in 18(d); the fluorescence imaging of HC-77 (Example 32) in mice with lung cancer cells A549 cells as shown in 18(e); the fluorescence imaging of HC-80 (Example 32) in mice with cholangiocarcinoma MCC cells as shown in 18(f). From Figure 18 It can be seen that the photosensitizing drugs HB-28 in basal cell carcinoma, HC-36 in squamous skin cancer, HB-45 in melanoma, HC-73 in cutaneous T-cell lymphoma, HC-77 in lung cancer, and HC-80 in cholangiocarcinoma have strong enrichment ability at the tumor site when injected intratumorally into tumor cells and tissues.

[0297] By changing the molecular structure and different substituents, various hypocrellin derivatives of the present invention also have similar enrichment effects on other different tumors. These derivatives have good drug enrichment in mice with esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, and bladder cancer.

[0298] In addition to having good enrichment in subcutaneous tumors of mice, what is the drug enrichment effect of the derivatives of this application at the site of orthotopic tumors in mice? To study the orthotopic enrichment of such derivatives in tumors, an orthotopic model of glioblastoma was obtained by inoculating orthotopic tumors in the brains of mice with glioblastoma C6 cells for fluorescence imaging. Using the derivative HB-3-PEG12 in Example 3 as a photosensitizing drug, it was injected into tumor-bearing mice through the tail vein at a dose of 10 mg / kg, and the fluorescence imaging behavior in tumor-bearing mice was observed. The fluorescence signals at the tumor site at 0, 2, 3.5, and 5 hours were collected using a small animal in vivo imaging system. As Figure 19 shown, as time prolongs, the fluorescence signal in the tumor area gradually increases and reaches the highest at 5 h, indicating that the photosensitizing drug HB-3-PEG12 is highly enriched at the tumor site, while there is no drug enrichment in the non-tumor area of the brain. The photosensitizing drug has very good targeted enrichment for glioblastoma. In addition, there is no fluorescence in the brains of blank mice without administration.

[0299] In the operation of glioma, there is no mediated drug for positioning to guide the resection of tumors. It is found that such derivatives can specifically accumulate in glioma tissues, and there is no photosensitizer accumulation in the brain regions without tumors, showing good tumor-targeted enrichment. At this time, irradiating the tumor tissue with light of a specific wavelength can excite detectable fluorescence to locate the position of the tumor tissue, which is used for fluorescence-guided glioma resection surgery; on the other hand, such derivatives can generate singlet oxygen under photosensitive conditions, resulting in damage to tumor cells and being used for photodynamic therapy of glioma, with basically no damage to normal tissues. The derivatives provided in this application not only have good enrichment effects on glioma, but also have good drug enrichment in mice with other tumors, such as esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, basal cell carcinoma, squamous cell skin cancer, melanoma, prostate cancer, and bladder cancer. The present invention discloses for the first time that a derivative of hypocrellin is used as a fluorescence-mediated material to guide the resection of solid tumors.

[0300] The above Figures 15-19 The above results show that the various derivatives provided in the present invention are used for fluorescence imaging of tumor-bearing mice. It can be seen that these photosensitive drugs produce strong or moderate fluorescence imaging at the tumors of mice, indicating that such derivatives can be used as photosensitive drugs for fluorescence imaging in tumor cells of tumor-bearing mice and can be used as fluorescence-mediated materials to guide the resection of tumor boundaries during surgery. The tumors include: esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioma, basal cell carcinoma, squamous cell skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, and bladder cancer. Other derivatives in this application also have in-situ enrichment effects on the above tumor cells, can be used for fluorescence imaging of the above tumor cells in vivo, and can be used as mediated materials to guide the resection of tumor boundaries during surgery. The fluorescence imaging results of some of the above hypocrellin derivatives in tumors are shown in Table 5.

[0301] Table 5: In-vivo enrichment of some derivatives of the present invention in digestive tract tumors, head and neck tumors, skin tumors, and genitourinary tumors

[0302]

[0303]

[0304] Example 46 Photodynamic therapy

[0305] In view of the fact that the hypocrellin derivatives of the present invention can efficiently kill various tumor cells in cell experiments and can be well enriched in mouse tumors as photosensitizing drugs, the inventors further tested such derivatives as photodynamic drugs to kill tumor cells in mice. Using mice as a model, tumors were inoculated in mice with A549 lung cancer cells. When the tumor volume reached about 200 mm 3 or so, a lung cancer animal model was obtained for photodynamic therapy. The photosensitizing drug HB-1-PEG8 (Example 1) at a dose of 10 mg / kg was injected into the tail vein of tumor-bearing mice, and the in vivo fluorescence imaging behavior was observed. The fluorescence signal at the tumor site was collected using a small animal in vivo imaging system for 4 h. After 4 h, the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 for 10 min, and the data were recorded. On the second day after photodynamic therapy for mouse lung cancer, scabs appeared, gradually shrank by the sixth day, and completely disappeared after the twelfth day, indicating that the lung cancer in mice was completely inhibited by photodynamic therapy ( Figure 20 ). For the control group of lung cancer-bearing mice (only injected with normal saline, without injecting the photosensitizing drug), the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 for 10 min. After 6 days, it was found that the tumor was growing rapidly, and after 12 days, the tumor was very serious, without the effect of inhibiting the tumor. Therefore, HB-1-PEG8, as a photosensitizing drug for photodynamic therapy of lung cancer, has an obvious effect of killing tumor cells and can inhibit tumor regeneration and recurrence.

[0306] The experiment also used a subcutaneous mouse model of glioma (inoculated with C6 cells) for photodynamic therapy. The photosensitizing drug HB-6-PEG6 (Example 6) at a dose of 10 mg / kg was injected into the tail vein of tumor-bearing mice, and the in vivo fluorescence imaging behavior was observed. After 4 h, the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 for 10 min, and the data were recorded. As Figure 21 (b), scabs appeared on the second day after photodynamic therapy for mouse glioma, gradually shrank by the sixth day, and completely disappeared after the fourteenth day, indicating that the growth of glioma in mice was completely inhibited by photodynamic therapy. For the control group of glioma mice (only injected with normal saline, without injecting the photosensitizing drug), under the same experimental conditions, the tumor was growing rapidly after 6 days, without any effect of inhibiting the tumor, Figure 21 (a). Therefore, HB-6-PEG6, as a photosensitizing drug for photodynamic therapy of glioma, has an obvious effect of killing tumor cells and can inhibit tumor regeneration and recurrence.

[0307] The experiment also used a subcutaneous mouse model of melanoma (inoculated with B16 cells) for photodynamic therapy. HC-9-PEG8 (Example 9) was injected into the tail vein of tumor-bearing mice, and in vivo fluorescence imaging was observed. After 4 h, the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 for 10 min, and the data were recorded. As shown in Figure 21 (c), scabbing occurred on the second day after photodynamic therapy for mouse melanoma, gradually shrank on the sixth day, and completely disappeared after the 14th day, indicating that the tumor in the mice was completely inhibited by photodynamic therapy. For the tumor-bearing mice in the control group, under the same conditions, but only normal saline was injected and no photosensitizing drug was injected. After 6 days, the tumor was growing rapidly and there was no effect of inhibiting the tumor. Therefore, HC-9-PEG8 used as a photosensitizing drug for photodynamic therapy of melanoma has the effect of killing tumors and can inhibit the regeneration and recurrence of tumors.

[0308] The experiment also used a subcutaneous mouse model of bladder cancer (inoculated with MBT-2 cells) for photodynamic therapy. HB-63 (Example 28) was injected into the tail vein of tumor-bearing mice, and in vivo fluorescence imaging was observed. After 4 h, the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 for 10 min, and the data were recorded. As shown in Figure 21 (d), scabbing occurred on the second day after photodynamic therapy for mouse bladder cancer, gradually shrank on the sixth day, and completely disappeared after the 14th day, indicating that the tumor in the mice was completely inhibited after photodynamic therapy. For the tumor-bearing mice in the control group, under the same experimental conditions, but only normal saline was injected and no photosensitizing drug was injected. After 6 days, it was found that the tumor was growing rapidly and there was no effect of inhibiting the tumor. Therefore, HB-63 used as a photosensitizing drug for photodynamic therapy of bladder cancer has an obvious effect of killing tumors and can inhibit tumor regeneration and recurrence.

[0309] H&E staining and pathological analysis were performed on the tumor tissues of mice after different treatments. It was found that the tumor cells in the blank control group were not damaged, and the tumor cells in the photodynamic therapy group were completely dead. Statistical analysis of the survival rate of mice found that compared with the control group, none of the mice treated with photodynamic therapy died within 30 days, indicating that photodynamic therapy has a good anti-tumor effect on tumor-bearing mice.

[0310] Taking mice as the model, the experiment also inoculated subcutaneous tumors of mice with cholangiocarcinoma (MCC cells) to obtain a cholangiocarcinoma model for photodynamic therapy. The photosensitizing drug HB-29 (Example 19) was injected into the tumor of tumor-bearing mice, and its fluorescence imaging behavior in tumor-bearing mice was observed. After 4 h, the tumor site was irradiated with a 635 nm laser at 0.1 W / cm 2 for 10 min, and the data were recorded ([ Figure 22a). Crusting occurred on the 2nd day after photodynamic therapy for cholangiocarcinoma in mice, gradually shrank on the 6th day, and completely disappeared after the 14th day, indicating that after photodynamic therapy, cholangiocarcinoma in mice was almost completely inhibited. For the control group of mice, under the same experimental conditions, only normal saline was injected without injecting photosensitizing drugs. After 6 days, it was found that the tumor was growing rapidly without any inhibitory effect on the tumor. Therefore, HB-29, as a photosensitizing drug, has an obvious tumor-killing effect and can inhibit the regeneration and recurrence of tumors in photodynamic therapy for cholangiocarcinoma.

[0311] The experiment also used a subcutaneous mouse model of colon cancer (inoculated with HCT116 cells) for photodynamic therapy. The photosensitizing drug HB-45 (Example 23) was injected into the tumor of tumor-bearing mice, and in vivo fluorescence imaging was observed. After 4 h, a 635 nm laser with 0.1 W / cm 2 irradiated the tumor site for 10 min. As Figure 22 (b), crusting occurred on the 2nd day after photodynamic therapy for colon cancer in mice, gradually shrank on the 6th day, and completely disappeared after the 14th day, indicating that photodynamic therapy completely inhibited colon cancer in mice. For the control group of mice, under the same experimental conditions, only normal saline was injected without injecting photosensitizing drugs. After 6 days, it was found that the tumor was growing rapidly without any inhibitory effect on the tumor. Therefore, HB-45, as a photosensitizing drug, has an obvious tumor cell-killing effect and can inhibit tumor regeneration and recurrence in photodynamic therapy for colon cancer.

[0312] The experiment used a subcutaneous mouse model of oral cancer (inoculated with CAL27 cells) for photodynamic therapy. The photosensitizing drug HC-73 (Example 31) was injected into the tumor of tumor-bearing mice, and in vivo fluorescence imaging was observed. After 4 h, a 635 nm laser with 0.1 W / cm 2 irradiated the tumor site for 10 min, and the data was recorded. As Figure 22 (c), crusting occurred on the 2nd day after photodynamic therapy for oral cancer in mice, gradually shrank on the 6th day, and completely disappeared after the 14th day, indicating that after photodynamic therapy, oral cancer in mice was completely inhibited. For the control group of mice, under the same experimental conditions, only normal saline was injected without injecting photosensitizing drugs. After 6 days, the tumor was growing rapidly without any inhibitory effect on the tumor. Therefore, HC-73, as a photosensitizing drug, has an obvious tumor-killing effect and can inhibit the regeneration and recurrence of tumors in photodynamic therapy for oral cancer.

[0313] The experiment used a subcutaneous mouse model of basal cell carcinoma (inoculated with BCC cells) for photodynamic therapy. The photosensitizing drug HC-80 (Example 32) was injected into the tumor of tumor-bearing mice, and in vivo fluorescence imaging was observed. After 4 h, a 635 nm laser with 0.1 W / cm 2 irradiated the tumor site for 10 min, and the data was recorded. As Figure 22(d), Scabbing occurred on the second day of photodynamic therapy, gradually shrank on the sixth day, and completely disappeared after 14 days, indicating that after photodynamic therapy, the tumors in the mice were completely inhibited. For the control group of mice, under the same experimental conditions, but only injected with normal saline and not injected with photosensitizing drugs, it was found that the tumors were growing rapidly after 6 days, without any effect of inhibiting tumors. Therefore, as a photosensitizing drug for photodynamic therapy of basal cell carcinoma, HC-80 has an obvious effect of killing tumors and can inhibit the regeneration and recurrence of tumors.

[0314] The above details illustrate that the hypocrellin derivatives of the present invention can efficiently kill various tumor cells in the body of tumor-bearing mice (such as lung cancer, cholangiocarcinoma, colon cancer, oral cancer cells, basal cell carcinoma cells, melanoma cells, glioma cells, bladder cancer cells. By injecting a certain dose of photosensitizing drug into the tumor-bearing mice and cooperating with laser irradiation of a certain intensity and wavelength, good photodynamic therapy effects have been achieved. Since such photosensitizing drugs have a killing effect on tumor cells, it is believed that for other tumor mice, such as esophageal cancer, gastric cancer, liver cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, squamous cell skin cancer, cutaneous T-cell lymphoma, prostate cancer, etc., good photodynamic therapy effects can also be obtained. Other compounds disclosed in the present invention, whether they are derivatives of hypocrellin or deacetyhypocrellin, connected with different substituted amino groups, all show good ability to inactivate tumor cells by photodynamic under photodynamic conditions. Therefore, the 2-position amino-substituted or ethylenediamine-substituted derivatives of hypocrellin disclosed in the present invention can all treat tumors in the body of mice well.

[0315] Comparative Example 1

[0316] The photodynamic effect of the hypocrellin derivative in the present invention on HeLa is as Figure 23 shown. Under red light irradiation of the photosensitizing drug, HB-1-PEG6 at a concentration of 200 nM can kill more than 80% of HeLa cells, and the half-lethal concentration IC 50 value is about 120 nM; similarly, HB-73 at a concentration of 200 nM can kill more than 80% of HeLa cells, and the half-lethal concentration IC 50 value is about 80 nM; HC-80 at a concentration of 200 nM can kill more than 80% of HeLa cells, and the half-lethal concentration IC 50 value is about 80 nM;

[0317] Under the same conditions, HB-1-PEG6 can kill 80% of esophageal cancer cells AKR, gastric cancer cells MFC, lung cancer cells A549, liver cancer cells HCC, cholangiocarcinoma cells MCC, and colon cancer cells HCT116 only at a concentration of 50 nM, and the half-lethal concentration IC 50The value is about 20-30 nM (Table 2); under the same conditions, HB-73 can kill 85% of head and neck cancer cells SCC2, brain cancer cells G442, tongue cancer cells TSCCa, nasal cancer cells KB, oral cancer cells CAL27, and glioma cells C6 at a concentration of only 50 nM, and the half-lethal concentration IC 50 The value is about 20-30 nM (Table 3); under the same conditions, HC-80 can kill 85% of basal cell carcinoma cells BCC, squamous skin cancer cells PECA, melanoma cells B16, prostate cancer cells LNCaP, and bladder cancer cells MBT-2 at a concentration of only 50 nM, and the half-lethal concentration IC 50 The value is about 20-30 nM (Table 4). Therefore, from Figure 23 Comparing with Tables 2-4, it can be seen that the phototoxic effect of the hypocrellin derivative disclosed in the present invention on the above-mentioned tumor cells is significantly higher than that on HeLa cells.

[0318] Obviously, the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, rather than limitations on the implementation manners of the present invention. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is impossible to list all the implementation manners here. Any obvious changes or modifications derived from the technical solutions of the present invention still fall within the protection scope of the present invention.

[0319] In addition, it should be noted that the hypocrellin derivatives to be protected in the present invention all contain two enol tautomers, and the chemical structures of the two isomers are as shown in Formula (I) and Formula (I’), which of course are within the protection scope. For the sake of simplicity, only one enol tautomer is listed in all the embodiments of the present invention, and the other enol tautomer and its corresponding general structural formula are described in detail in the specification, and its structure is of course within the protection scope. In addition, the general structural formula of the hypocrellin derivatives involved in the present invention contains a polyethylene glycol unit (PEGn), and the unit number n is any integer between 1 and 50, and the corresponding chemical structures are of course within the protection scope. For the sake of simplicity, only some integers are listed in all the embodiments of the present invention, and the corresponding general structural formulas of the rest are described in detail in the specification, and their structures are of course within the protection scope. Any range recorded in the present invention includes the end values and any numerical values between the end values, as well as any sub-ranges formed by the end values or any numerical values between the end values.

[0320] The above describes the implementation manners of the present invention. However, the present invention is not limited to the above implementation manners. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of a hypocrellin derivative or a mixture thereof in the preparation of a photodynamic anti-tumor drug, wherein, the tumor is esophageal cancer, gastric cancer, lung cancer, cholangiocarcinoma, brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell skin cancer, cutaneous T-cell lymphoma, prostate cancer, bladder cancer, and the hypocrellin derivative is a compound represented by formula (II), its isomer or a pharmaceutically acceptable salt; The compound represented by formula (II) is the following specific compound: n is an integer between 1 and 100.

2. The use according to claim 1, characterized in that, the derivatives in formula II and formula II' are enol tautomers; 3. The use according to claim 1 or 2, characterized in that, the cancer cells corresponding to the tumor are: esophageal cancer cells, gastric cancer cells, lung cancer cells, cholangiocarcinoma cells, head and neck cancer cells, brain cancer cells, tongue cancer cells, nasal cancer cells, oral cancer cells, glioblastoma cells, basal cell carcinoma cells, squamous cell skin cancer cells, cutaneous T-cell lymphoma cells, prostate cancer cells, bladder cancer cells.

4. The use according to claim 1 or 2, characterized in that, the drug is a photodynamic drug, a fluorescence-mediated drug.

5. Use of a compound as shown below, its isomer or a pharmaceutically acceptable salt in the preparation of a fluorescence-mediated drug for guiding the boundary resection of tumors; the tumors are esophageal cancer, gastric cancer, lung cancer, cholangiocarcinoma, brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer, glioblastoma, basal cell carcinoma, squamous cell skin cancer, cutaneous T-cell lymphoma, prostate cancer, bladder cancer; n is an integer between 4 and 100.

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