Application of hypocrellin 2-site amino substituted or ethylenediamine substituted derivative in preparation of antitumor photodynamic drugs

Abstract

The invention relates to a hypocrellin 2-site amino substituted derivative (formula I) or ethylenediamine substituted derivative (formula II and formula III) as a photodynamic drug for treating the following tumors: esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer and colon cancer cells related to digestive tract tumors; brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer and brain glioma cells related to head, neck and face tumors; basal cell carcinoma, squamous skin carcinoma, skin T cell lymphoma and melanoma cells related to skin tumors; prostate cancer and bladder cancer cells related to genitourinary system tumors. Meanwhile, the derivative can locate the position of a tumor tissue through self-fluorescence and is used for fluorescence-guided tumor operation excision.

Description

[0001] This application is a divisional application of the patent application No. 202111372435.5, filed on November 18, 2021, with the State Intellectual Property Office of China, and entitled "Application of a derivative of amine or ethylenediamine substituted at position 2 of bambmerin in preparation of an anti-tumor photodynamic drug". TECHNICAL FIELD

[0002] The present application relates to the technical field of photosensitizer drugs, in particular to application of a bambmerin derivative in preparation of a photodynamic anti-tumor drug. BACKGROUND

[0003] Photodynamic action is that under the action of light, photosensitizer causes functional or morphological changes of body cells or biological molecules, and then causes cell damage and necrosis. The non-invasive or minimally invasive treatment technology for selectively destroying diseased tissues to achieve the treatment purpose is called photodynamic therapy (PDT). In photodynamic therapy, photosensitizer selectively accumulates in target cell tissues, is irradiated by light of appropriate wavelength, and absorbs photon energy to change from ground state to excited state. Because the excited state of photosensitizer is extremely unstable, it will subsequently release energy back to the ground state through physical deactivation or chemical deactivation process. Among them, the chemical deactivation process produces a large number of free radicals and singlet active oxygen (ROS), which oxidize a variety of biological macromolecules such as amino acids, unsaturated fatty acids, adenosine, etc. through cell production, damage cell structure, affect cell function, and then cause cell death. The photosensitizer absorbed by normal tissues has been metabolized and excreted, and does not produce photodynamic action. Photosensitizer, light source and oxygen are the three elements of photodynamic therapy. The specific accumulation of photosensitizer in the target, the chemical action of light and photosensitizer to produce singlet oxygen and other active substances, and the directional damage of active substances to target cells are the key links of photodynamic therapy.

[0004] Tumor is the most widely used field of photodynamic therapy. Currently, there are more than ten kinds of photodynamic therapy for tumors in clinical practice, which have achieved good results. PDT can cure early-stage malignant tumors in situ, and can be used for palliative treatment of intermediate tumors to improve symptoms and prolong life. PDT can be used for the treatment of tumor resection sites, and laser irradiation can prevent tumor recurrence. Malignant tumors of the body surface are particularly suitable for photodynamic therapy because the lesion is shallow and the laser can directly penetrate the lesion. For malignant tumors in deeper parts, endoscopy can be used for light treatment. In addition, through some special methods, photodynamic therapy can also be used for interventional treatment of liver cancer or bone marrow purification. Compared with traditional treatment methods (surgery, chemotherapy, and radiotherapy), photodynamic therapy is more targeted and can selectively eliminate primary and recurrent tumors with less toxicity and less damage to normal cells. It can treat various types of tumors. With the improvement of photosensitizers and the development of photodynamic therapy technology, the limitations and deficiencies of PDT are gradually improving, but its application in clinical practice still has certain limitations. For different patients, how to develop individualized treatment plans, determine the treatment range according to the size and depth of the tumor, and select the light source and power size need further research. At present, photodynamic therapy has been reported in the treatment of superficial tumors, 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 (brain glioma), reproductive tumors (prostate cancer, bladder cancer, cervical cancer, etc.), and digestive system tumors (cholangiocarcinoma, gastric cancer, lung cancer, liver cancer, colorectal cancer, etc.). Photodynamic therapy not only has good efficacy for many primary tumors, but also has unique technical advantages for many metastatic tumors. Studies have shown that the possible mechanism of photodynamic therapy in anti-tumor is direct killing of tumor cells or anti-vascular photodynamic therapy. Direct killing of tumor cells takes advantage of the dual selectivity of photodynamic therapy, selective retention of photosensitizers in tumor cells, and the generation of ROS near the lesion under the irradiation of specific wavelength light, leading to tumor cell lesions and death. Anti-vascular photodynamic therapy is a method in which tumor cell survival depends on blood supply. Photodynamic therapy can damage the blood vessels associated with tumors, leading to ischemic death of the tumor. If the concentration of photosensitizers in the blood vessels is at the peak value, light irradiation can cause damage to the microvessels, leading to insufficient blood supply in the lesion, and thus causing cell necrosis or apoptosis.

[0005] Photosensitizer is the most critical factor in photodynamic therapy. The first generation of photosensitizer developed in the 1970s and early 1980s, mainly porphyrin derivatives (HpD) photosensitizer, with porphyrin as a representative, a mixture extracted from pig or cow blood, its main effective component is double blood porphyrin ether or ester. Its clinical application history is the longest and the research is the most detailed. Including Canada, the United States, Germany, Russia, Belgium and our own research of cancer porphyrin, cancer light porphyrin, light porphyrin, etc. In the past 30 years, there are many HpD commercial products for clinical use, and thousands of patients have received PDT treatment. The research results of HpD have achieved success in the treatment of some body surface tumors, and the treatment effect on respiratory tract tumors such as bronchogenic carcinoma is also quite significant. In addition, HpD has good effect on the treatment of upper gastrointestinal malignant tumors, and even a few cases have reached clinical cure. Barrett's esophagus is a very important precancerous lesion of esophageal cancer, and HpD has become one of the preferred treatment methods. In addition, HpD has good therapeutic effect on head and neck tumors, brain tumors, bladder cancer, and bile duct cancer. The first generation of photosensitizer has definite effect in tumor treatment, but still has many shortcomings, such as complex composition, poor tissue selectivity, slow metabolism, long light-avoiding time, certain toxicity, etc. The second generation of photosensitizer developed in the late 1980s, the activity, absorption spectrum and tissue selectivity of the second generation of photosensitizer have been greatly improved compared with the first generation of photosensitizer. The second generation of photosensitizer is mostly monomer compound, including porphyrin derivatives, metal phthalocyanine, chlorophyll degradation derivatives, fused ring quinone compounds and porphine, etc. In 1990, endogenous porphyrin photosensitizer ALA has been successfully applied to the treatment of genital condyloma acuminatum, and now ALA mediated photodynamic therapy has been widely used in tumor skin diseases such as squamous cell carcinoma and basal cell carcinoma; Hypericin photosensitizer has been studied in anti-tumor in recent years, and hypericin mediated PDT can treat various tumors such as pancreatic cancer, bladder cancer, lymphoma, prostate cancer and basal cell carcinoma; Phthalocyanine photosensitizer has been applied in Russia and other countries and regions, and its anti-tumor and anti-infection effect is significant and also shows good safety in phase I / II clinical study. The chemical structure of the second generation of photosensitizer is clear, the yield of singlet oxygen is high, the photosensitive period is short, and the maximum absorption wavelength is red shifted, which increases the depth of photodynamic therapy, so the commercialization and clinical application prospect is very optimistic, but there are still some defects such as high difficulty in separation and purification and unsatisfactory targeting. The third generation of photosensitizer developed in the late 20th century, in order to improve the biocompatibility, targeting and develop photosensitive drug delivery system, the third generation of photosensitizer is to combine porphyrin or phthalocyanine with some chemicals with biological properties, such as amino acids, polymers, proteins, sugars, liposomes, tumor tissue expressed antigens, receptors corresponding antibodies or ligands, etc. to construct a photosensitive system that can both target tumor and play the effect of photodynamic therapy.For example, hematoxylin combined with monoclonal antibodies has a very large killing effect on target cells; phthalocyanine-lipid protein complexes have a greater uptake rate in tumor tissues than uncomplexed phthalocyanine complexes, both in vitro and in vivo experiments. The third generation of photosensitizers is still in the preclinical animal research stage, and there is still a long way to go before it can be used in clinical applications.

[0006] Although the above photosensitizers have undergone decades of development in different periods, and have made great progress in photodynamic therapy of tumors, the types of such photosensitizers are relatively single, and basically all are derivatives of porphyrin. The metabolic time of porphyrin derivatives in the body is relatively long, and the metabolic time of early HpD in the human body needs 1-3 months. Although the metabolic time of later developed huaporphyrin in the body is greatly shortened to one or two weeks, there are still problems such as weak light absorption ability of the phototherapy window, difficulty in chemical separation of stereoisomers, and therefore there is an urgent need to develop high-efficiency photosensitizers with new structures.

[0007] Hypocrellins are natural photosensitizers extracted and separated from the stroma of Hypocrella bambuase and Shiraiabambusicola, a bamboo parasitic fungus found in the bamboo forest above 3000 meters in Yunnan, China, and belong to perylene quinone compounds. Natural hypocrellins mainly include hypocrellin A, hypocrellin B and other components, among which hypocrellin A accounts for 95%. Under alkaline conditions, hypocrellin A can be dehydrated to convert to hypocrellin B, and the conversion rate can be as high as 99%. Compared with the phototherapy drug hematoxylin derivative (HpD) used in clinical practice, hypocrellins have many advantages, such as simple composition, easy purification of raw materials, high triplet state quantum yield and singlet oxygen quantum yield, high phototoxicity, low dark toxicity, dual photodynamic mechanism of Type I and Type II, fast excretion, etc. It is a very promising photosensitizer. In addition, the structure of hypocrellin is easy to modify, and the modified derivatives can meet the requirements of strong light absorption ability in the phototherapy window (600-900 nm) and water-soluble performance meeting the needs of clinical intravenous injection, so hypocrellins have broad application prospects as photodynamic drugs. However, hypocrellin photosensitizers for tumor treatment are still in the laboratory research stage, and there is no related hypocrellin drug for clinical research. Therefore, there is an urgent need to develop new hypocrellin-based photodynamic drugs for photodynamic therapy of tumors in clinical practice.

[0008] The inventors have disclosed a derivative of 2-amino or ethylenediamine substituted of shiitake mushroom red pigment (CN201610894129.0) for the first time. After 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, bile duct cancer, colon cancer cells related to digestive system tumors; head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, brain glioma cells related to head and neck tumors; basal cell carcinoma, squamous skin cancer, cutaneous T-cell lymphoma, melanoma cells related to skin tumors; prostate cancer, bladder cancer cells related to genitourinary system tumors. In addition, for some special tumors (such as brain glioma), there is no problem of guiding the resection of tumors during surgery without the mediation of drugs. The inventors found that such shiitake mushroom red pigment derivatives can specifically accumulate in brain glioma cells and tissues, and there is no photosensitizer accumulation in the brain area without tumors, which has good tumor targeting enrichment effect. At this time, the tumor tissue can be irradiated with a specific wavelength of light to excite detectable fluorescence to locate the position of the tumor tissue, which is used for fluorescence-guided tumor resection surgery (FGS). SUMMARY

[0009] To improve the above technical problems, the present application provides a compound as shown in formula (I), formula (II) or formula (III), a derivative of shiitake mushroom red pigment 2-amino or ethylenediamine substitution, its isomer, isotopically labeled, pharmaceutically acceptable salt or solvate in the preparation of photodynamic antitumor drugs, the tumor is esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, brain glioma, basal cell carcinoma, squamous skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer.

[0010]

[0011] The structural formula of R1 in formula (I) is shown in formula (IV), and R2 is -H or -COCH3:

[0012]

[0013] In formula (IV), 0≤m≤12, 0≤n≤500, 0≤p≤12, 0≤q≤12; m, n, p, q are zero or positive integers; Y is a linking group; Z is an end group; (OCH2CH2) n is a polyethylene glycol unit;

[0014] In formula (IV), the linking group Y is O, NH, S, carboxylate, amide, sulfocarboxylate, phenylene, 3-12 carbon atoms alkenylene or 3-12 carbon atoms cycloalkyl;

[0015] The 3-12 carbon atom cycloalkyl group includes substituted or unsubstituted cycloalkyl groups or cycloalkyl groups containing heteroatoms, such as oxygen, nitrogen or sulfur atoms; the substituents are alkyl groups of 1-12 carbon atoms;

[0016] The end group Z in formula (IV) is hydrogen, an alkyl group of 1-12 carbon atoms, an alkoxy group of 1-12 carbon atoms, phenyl, hydroxyl, amino, mercapto, carboxylic acid group, sulfonic acid group, pyridyl, quaternary ammonium salt or pyridine salt;

[0017] When the end group Z is a quaternary ammonium salt, the three substituents on the quaternary ammonium salt are independently or simultaneously alkyl groups of 1-12 carbon atoms; the anion in the quaternary ammonium salt is an anion allowed in pharmaceutical preparations;

[0018] When the end group Z is a pyridine salt, the substituents on the pyridine ring are in the ortho, meta or para position; the pyridine salt is formed by quaternization of pyridine with halogenated hydrocarbons containing 1-12 carbon atoms of different chain lengths; the anion in the pyridine salt is an anion allowed in pharmaceutical preparations;

[0019] The substituent R2 in formula (II) is -H or -COCH3:

[0020] R3-R8 in formula (II) are the same or different, as defined for the substituent R1 in formula (I), i.e. R3-R8 are the same or different and independently as shown in formula (IV).

[0021] Preferably, the linking group Y in formula (IV) is: -O-; -NH-; -S-; -COO-; -O-CO-; -CONH-; -NH-CO-; -SO3-; -SO2-NH-; -C6H4- (phenyl); -C3H4- (cyclopropyl); -C4H6- (cyclobutyl); -C5H8- (cyclopentyl); -C5H7(CH3)- (methylcyclopentyl); -C6H 10 (cyclohexyl); -C6H9(CH3)- (methylcyclohexyl); -C7H 12 (cycloheptyl); (piperazinyl).

[0022] Preferably, the end group Z in formula (IV) is: -H; -CH3; -C2H5; -C4H9; -C6H 13 ; -OCH3; -OC2H5; -OC4H9; -OC6H 13 ; -C6H5; -OH, -NH2; -SH; -COOH; -COOCH3; -SO3H; -C5H4N; -C5H4N + ; -N + (CH3)3; -N + (C2H5)3; -N + (C6H13 )3; -N + (CH3)2(C2H5); -N + (CH3)2(C6H 13 ); -N + (CH3)2(C8H 17 ).

[0023] 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, and q is an integer from 0 to 8, such as 0, 1, 2, 3, 4, etc.

[0024] In particular, the derivatives of formula I and I' are enol tautomers; the derivatives of formula II and II' are enol tautomers; the derivatives of formula III and III' are enol tautomers.

[0025]

[0026] Preferably, the substituent R1is an alcohol of different chain length, and a carboxylate with carboxyl polyethylene glycol: -(CH2) m -OH; -(CH2) m -OCH3; -(CH2) m -O-CO-CH2CH2-(OCH2CH2) n -OCH3[m is an integer from 1 to 8, n is an integer from 0 to 100];

[0027] Preferably, the substituent R1is a carboxylic acid of different chain length, and a carboxylate or amide with polyethylene glycol: -(CH2) m -COOH; -(CH2) m -COOCH3; -(CH2) m -CO-(OCH2CH2) n -OH; -(CH2) m -CO-(OCH2CH2) n -OCH3; -(CH2) m -CO-NH-CH2CH2-(OCH2CH2) n -OCH3[m is an integer from 1 to 8, n is an integer from 0 to 100];

[0028] Preferably, the substituent R1is a sulfonic acid group of different chain length, and a sulfonate or sulfonamide with polyethylene glycol: -(CH2) m -SO3H; -(CH2) m -SO2-(OCH2CH2) n -OH; -(CH2) m-SO2-(OCH2CH2) n -OCH3; -(CH2) m -SO2-NH-CH2CH2-(OCH2CH2) n -OH; -(CH2) m -SO2-NH-CH2CH2-(OCH2CH2) n -OCH3[m is an integer between 1 and 8, n is an integer between 0 and 100];

[0029] Preferably, said substituent R1 is a thio-polyethylene glycol: -CH2CH2-SH; -CH2CH2-S-CH2CH2OH; -CH2CH2-S-CH2CH2OCH3; -CH2CH2-S-CH2CH2-(OCH2CH2) n -OH;

[0030] Preferably, said substituent R1 is an alkyl, amino, hydroxyl, or a substituent containing a phenyl, pyridyl, olefin: -H; -CH3; -C2H5; -C3H7; -C4H9; -C5H 11 ; -C6H 13 ; -C8H 17 ; -NH2; -NHCH3; -NHC2H5; -OH; -CH2CH=CH2; -(CH2)2CH=CH2; -(CH2)3CH=CH2; -CH2C6H5; -C5H4N; -CH2C5H4N; -(CH2)2C5H4N; -NHC6H5; -NHC5H4N;

[0031] Preferably, said substituent R1 is a cycloalkyl-containing substituent: -C3H5 (cyclopropyl), -C4H7 (cyclobutyl), -C5H9 (cyclopentyl), -C6H 11 (cyclohexyl), -C6H 10 (CH3) (methylcyclohexyl), -C6H 10 (OH) (hydroxycyclohexyl), -C7H 13 (cycloheptyl), -CH2C6H 10 COOH, -CH2C6H 10 COOCH3, -CH2C6H 10 OH, -C6H 10 COOH;

[0032] More preferably, said substituent R1 is a cyclohexane containing substituent: -C6H 10 -OH; -CH2C6H 10 COOH; -CH2C6H 10 COOCH3; -CH2C6H10 OH; -C6H 10 COOH), said substituents being in ortho, para, meta position with respect to the cyclohexane ring;

[0033] Preferably, said substituent R1 is a substituent containing a quaternary ammonium salt: -(CH2) m -N + (CH3)3; -(CH2) m -N + (CH3)2(C2H5); -(CH2) m -N + (CH3)2(C3H7); -(CH2) m -N + (CH3)2(C4H9); -(CH2)3-N + (CH3)2(C6H 13 ); -(CH2) m -N + (CH3)2(C8H 17 ); -(CH2) m -N + (CH3)2(C 12 H 25 ); -(CH2) m -O-CO-(CH2)2-N + (CH3)3; -(CH2) m -O-CO-(CH2)3-N + (CH3)3; -(CH2) m -O-CO-(CH2)4-N + (CH3)3; -(CH2) m -O-CO-(CH2)5-N + (CH3)3; -(CH2) m -O-CO-(CH2)6-N + (CH3)3; -(CH2) m -COO-(CH2)2-N + (CH3)3; -(CH2) m -COO-(CH2)3-N + (CH3)3; -(CH2) m -COO-(CH2)4-N + (CH3)3; -(CH2) m -COO-(CH2)5-N + (CH3)3; -(CH2) m -COO-(CH2)6-N + (CH3)3; -(CH2)m -CONH-(CH2)2-N + (CH3)3; -(CH2) m -CONH-(CH2)3-N + (CH3)3; -(CH2) m -CONH-(CH2)4-N + (CH3)3[m is an integer between 1 and 8];

[0034] Preferably, the substituent R1 is a heterocycle-containing substituent:

[0035] According to an embodiment of the application, the tumor is an esophageal cancer, a gastric cancer, a lung cancer, a liver cancer, a cholangiocarcinoma, a colon cancer, a head and neck cancer, a brain cancer, a tongue cancer, a nose cancer, an oral cancer, a brain glioma, a basal cell carcinoma, a squamous skin cancer, a cutaneous T-cell lymphoma, a melanoma, a prostate cancer, a bladder cancer.

[0036] According to an embodiment of the application, the tumor cell is an esophageal cancer cell AKR, a gastric cancer cell MFC, a lung cancer cell A549, a liver cancer cell HepG2, a cholangiocarcinoma cell MCC, a colon cancer cell HCT116, a head and neck cancer cell SCC2, a brain cancer cell G442, a tongue cancer cell TSCCa, a nose cancer cell KB, an oral cancer cell CAL27, a brain glioma cell C6, a basal cell carcinoma cell BCC, a squamous skin cancer cell PECA, a cutaneous T-cell lymphoma HH, a melanoma cell B16, a prostate cancer cell LNCaP, a bladder cancer cell MBT-2.

[0037] According to an embodiment of the application, the drug can be a photodynamic drug, a fluorescence-mediated drug.

[0038] According to an embodiment of the application, the drug can be enriched in the tumor cell.

[0039] According to an embodiment of the application, the pharmaceutically acceptable salt includes a salt formed after esterification of the compound of formula (I) with 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, with an inorganic base, including sodium, potassium, calcium, aluminum and ammonium salts, or with an organic base, including methylamine, ethylamine and ethanolamine salts, or esterification of the compound of formula (I) with a basic amino acid selected from lysine, arginine and ornithine, followed by formation of a salt with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid and phosphoric acid, or with an organic acid selected from formic acid, acetic acid, picric acid, methanesulfonic acid.

[0040] The present application also provides a use of a compound represented by formula (I), formula (II) or formula (III), isomers, isotope-labeled compounds, pharmaceutically acceptable salts or solvates thereof in the preparation of a fluorescently mediated drug for guiding the resection of tumor boundaries.

[0041] The present application also provides a use of a compound represented by formula (I), formula (II) or formula (III), isomers, isotope-labeled compounds or pharmaceutically acceptable salts thereof in the treatment of tumor diseases, 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, brain glioma, basal cell carcinoma, squamous skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer.

[0042] According to an embodiment of the present application, the treatment is the photodynamic inactivation of tumor cells or the fluorescently mediated drug guiding the resection of tumor boundaries.

[0043] The present application 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 a compound represented by formula (I), formula (II) or formula (III), isomers, isotope-labeled compounds, pharmaceutically acceptable salts or solvates thereof.

[0044] In some embodiments, the patient is a human.

[0045] Advantages

[0046] 1) The compound of formula (I), formula (II) or formula (III) according to the present application can efficiently kill the following cancer or tumor cells as a photodynamic drug: esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, brain glioma, basal cell carcinoma, squamous skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer. A concentration of 20-30 nM of such a photodynamic drug can kill more than 90% of tumor cells, while having little effect on normal cells, and the drug is basically excreted from the body after one week;

[0047] 2) The present application discloses for the first time a shiitake mushroom derivative as a mediated drug for guiding the resection of tumor in clinical tumor surgery. Such a derivative can specifically accumulate in tumor tissue, and there is no photosensitizer accumulation in the area without tumor, which has a good tumor targeting and enrichment effect. When the tumor tissue is irradiated with light of a specific wavelength, it can excite the production of detectable fluorescence to locate the position of the tumor tissue, which is used for fluorescently guided tumor resection surgery. BRIEF DESCRIPTION OF DRAWINGS

[0048] The specific embodiments of the present application will be further described in detail below with reference to the accompanying drawings.

[0049] Figure 1A general structure of the derivatives of the 2-amino or ethylenediamine substituted derivatives of hypocrellin B.

[0050] Figure 2 A synthetic method of the derivative HB-4-PEGn of the 2-polyethylene glycol-amino pentanol substituted derivative of hypocrellin B.

[0051] Figure 3 HC-74 and HC-75 are the reaction products of deacetylhypocrellin HC and methylethylenediamine.

[0052] Figure 4 (a) is a comparison chart of the absorption spectra of the commercial porphyrin photosensitizer PpIX and Ce6. Figure 4 (b) is a comparison chart of the absorption spectra of hypocrellin B HB, the 2-substituted product HB-6-PEG1 (Example 6), and the ethylenediamine substituted product HC-74 (Example 31).

[0053] Figure 5 (a) is an ESR chart of the reaction of the derivatives HB-45, HC-45 (Example 23) with active oxygen trapping agents: (a) singlet oxygen trapping agent; (b) superoxide radical trapping agent.

[0054] Figure 6 (a) is a photodegradation curve of HB-73 in Example 31. Figure 6 (b) is a photodegradation curve of HC-73 in Example 31. Figure 6 (c) is a photodegradation curve of HC-80 in Example 32.

[0055] Figure 7 (a) is a comparison chart of the photostability of the 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 20 mW / cm 2 of laser light for 30 min.

[0056] Figure 8 (a) is a comparison chart of the pH stability of the derivatives HB-3-PEG4 (Example 3), HC-3-PEG8 (Example 3), HC-73 (Example 31), HB-77 (Example 32), and commercial photosensitizers (hematoporphyrin HpD).

[0057] Figure 9 (a) is a dark field and bright field superimposed image; (b) is a dark field image; (c) is a bright field image.

[0058] Figure 10-1Dark toxicity plot (a) and phototoxicity plot (b) of esophageal cancer cell AKR for HB-1-PEG4 and HC-1-PEG8 (Example 1), and commercial photosensitizer HpD.

[0059] Figure 10-2 Dark toxicity plot (a) and phototoxicity plot (b) of lung cancer cell A549 for HB-3-PEG8 and HC-3-PEG16 (Example 3), and commercial photosensitizer HpD.

[0060] Figure 10-3 Dark toxicity plot (a) and phototoxicity plot (b) of liver cancer cell HepG2 for HB-6-PEG2 and HC-6-PEG6 (Example 6), and commercial photosensitizer HpD.

[0061] Figure 10-4 Dark toxicity plot (a) and phototoxicity plot (b) of colon cancer cell HCT116 for HB-14-PEG6 and HC-14-PEG12 (Example 13), and commercial photosensitizer HpD.

[0062] Figure 10-5 Dark toxicity plot (a) and phototoxicity plot (b) of biliary cancer cell MCC for HB-19-PEG4 and HC-19-PEG8 (Example 15), and commercial photosensitizer HpD.

[0063] Figure 10-6 Dark toxicity plot (a) and phototoxicity plot (b) of gastric cancer cell MFC for HB-45 and HC-45 (Example 23), and commercial photosensitizer HpD.

[0064] Figure 11 (a) Phototoxicity plot of brain cancer cell G442 for HB-10-PEG4 and HC-10-PEG8 (Example 10), and HpD;

[0065] Figure 11 (b) Phototoxicity plot of head and neck cancer cell SCC2 for HB-12-PEG8 and HC-12-PEG12 (Example 12), and HpD;

[0066] Figure 11 (c) Phototoxicity plot of tongue cancer cell TSCCa for HB-20-PEG6 and HC-20-PEG12 (Example 16), and HpD.

[0067] Figure 12 (a) Phototoxicity plot of nose cancer cell KB for HB-29 and HC-29 (Example 19), and commercial photosensitizer HpD;

[0068] Figure 12(b) is a plot of the phototoxicity of HB-61 and HC-61 (Example 28), and HpD on oral cancer cells CAL27;

[0069] Figure 12 (c) is a plot of the phototoxicity of HB-73 (Example 31), HC-80 (Example 32), and HpD on brain glioma cells C6.

[0070] Figure 13 (a) is a plot of the phototoxicity of HB-4-PEG8 and HC-4-PEG16 (Example 4), HpD on basal cell carcinoma cells BCC;

[0071] Figure 13 (b) is a plot of the phototoxicity of HB-9-PEG4 and HC-9-PEG8 (Example 9), and HpD on melanoma cells B16;

[0072] Figure 13 (c) is a plot of the phototoxicity of HB-36 and HC-36 (Example 20), and HpD on squamous skin carcinoma cells PECA.

[0073] Figure 14 (a) is a plot of the phototoxicity of HB-70 and HC-71 (Example 30), and HpD on prostate cancer cells LNCaP;

[0074] Figure 14 (b) is a plot of the phototoxicity of HB-73 and HC-73 (Example 31), and HpD on bladder cancer cells MBT-2;

[0075] Figure 14 (c) is a plot of the phototoxicity of HC-80 (Example 32) and HC-81 (Example 33), and HpD on skin T-cell lymphoma cells HH.

[0076] Figure 15 Fluorescence imaging of mice in different tumor models after tail vein injection of the derivatives of the present application for 4 h: (a) HB-1-PEG4 (Example 1) for esophageal cancer tumor AKR cell imaging; (b) HC-3-PEG12 (Example 3) for gastric cancer MFC cell imaging; (c) HB-6-PEG8 (Example 6) for lung cancer A549 cell imaging; (d) HB-7-PEG16 (Example 7) for liver cancer HepG2 cell imaging; (e) HC-9-PEG10 (Example 9) for cholangiocarcinoma MCC cell imaging; (f) HC-1-PEG50 (Example 1) for colon cancer HCT116 cell imaging.

[0077] Figure 16Fluorescent imaging of mice in different tumor models 4h after tail vein injection of the derivatives of the present application: (a) HB-11-PEG6 (Example 11) for brain cancer G442 cell imaging; (b) HC-18-PEG8 (Example 15) for head and neck cancer SCC2 cell imaging; (c) HC-45 (Example 23) for tongue cancer TSCCa cell imaging; (d) HC-57 (Example 27) for oral cancer CAL27 cell imaging; (e) HB-65 (Example 29) for nose cancer KB cell imaging; (f) HC-81-PEG16 (Example 34) for brain glioma C6 cell imaging.

[0078] Figure 17 Fluorescent imaging of mice in different tumor models 4h after tail vein injection of the derivatives of the present application: (a) HB-11-PEG6 (Example 11) for brain cancer G442 cell imaging; (b) HC-18-PEG8 (Example 15) for head and neck cancer SCC2 cell imaging; (c) HC-45 (Example 23) for tongue cancer TSCCa cell imaging; (d) HC-57 (Example 27) for oral cancer CAL27 cell imaging; (e) HB-65 (Example 29) for nose cancer KB cell imaging; (f) HC-81-PEG16 (Example 34) for brain glioma C6 cell imaging.

[0079] Figure 18 Fluorescent imaging of mice in different tumor models 4h after intratumoral injection of the derivatives of the present application: (a) HB-11-PEG6 (Example 11) for brain cancer G442 cell imaging; (b) HC-18-PEG8 (Example 15) for head and neck cancer SCC2 cell imaging; (c) HC-45 (Example 23) for tongue cancer TSCCa cell imaging; (d) HC-57 (Example 27) for oral cancer CAL27 cell imaging; (e) HB-65 (Example 29) for nose cancer KB cell imaging; (f) HC-81-PEG16 (Example 34) for brain glioma C6 cell imaging.

[0080] Figure 19 Fluorescent imaging of mice in different tumor models 4h after tail vein injection of the derivatives of the present application: (a) HB-11-PEG6 (Example 11) for brain cancer G442 cell imaging; (b) HC-18-PEG8 (Example 15) for head and neck cancer SCC2 cell imaging; (c) HC-45 (Example 23) for tongue cancer TSCCa cell imaging; (d) HC-57 (Example 27) for oral cancer CAL27 cell imaging; (e) HB-65 (Example 29) for nose cancer KB cell imaging; (f) HC-81-PEG16 (Example 34) for brain glioma C6 cell imaging.

[0081] Figure 20 For 0.1 W / cm 2The photos of the photodynamic therapy effect of the mice inoculated with lung cancer cells (A549) after 6 days and 12 days of photodynamic therapy by irradiation of 635 nm laser for 10 min: the first row is the mice without injection of drugs, only injection of normal saline; the second row is the mice injected with drug HB-1-PEG8 (Example 1) through tail vein.

[0082] Figure 21 Different photosensitizing drugs were injected through tail vein, and then the tumor sites of the tumor-bearing mice after 6 days of photodynamic therapy by irradiation of 0.1 W / cm2 of 635 nm laser for 10 min were photographed: Fig. (a) is the mice inoculated with brain glioma cells (C6) without injection of drugs, only injection of normal saline; Fig. (b) is the mice inoculated with brain glioma cells (C6) injected with drug HB-6-PEG6 (Example 6) through tail vein; Fig. (c) is the mice inoculated with melanoma cells (B16) injected with drug HC-9-PEG8 (Example 9) through tail vein; Fig. (d) is the mice inoculated with bladder cancer cells (MBT-2) injected with drug HB-63 (Example 28) through tail vein. 2 2 Different photosensitizing drugs were injected through tail vein, and then the tumor sites of the tumor-bearing mice after 6 days of photodynamic therapy by irradiation of 0.1 W / cm2 of 635 nm laser for 10 min were photographed: Fig. (a) is the mice inoculated with brain glioma cells (C6) without injection of drugs, only injection of normal saline; Fig. (b) is the mice inoculated with brain glioma cells (C6) injected with drug HB-6-PEG6 (Example 6) through tail vein; Fig. (c) is the mice inoculated with melanoma cells (B16) injected with drug HC-9-PEG8 (Example 9) through tail vein; Fig. (d) is the mice inoculated with bladder cancer cells (MBT-2) injected with drug HB-63 (Example 28) through tail vein.

[0083] Figure 22 Different photosensitizing drugs were injected through tail vein, and then the tumor sites of the tumor-bearing mice after 6 days of photodynamic therapy by irradiation of 0.1 W / cm2 of 635 nm laser for 10 min were photographed: Fig. (a) is the mice inoculated with brain glioma cells (C6) without injection of drugs, only injection of normal saline; Fig. (b) is the mice inoculated with brain glioma cells (C6) injected with drug HB-6-PEG6 (Example 6) through tail vein; Fig. (c) is the mice inoculated with melanoma cells (B16) injected with drug HC-9-PEG8 (Example 9) through tail vein; Fig. (d) is the mice inoculated with bladder cancer cells (MBT-2) injected with drug HB-63 (Example 28) through tail vein.

[0084] Figure 23 The photos of the photodynamic effect of derivatives HB-1-PEG6 (Example 1), HB-73 (Example 31), and HC-80 (Example 32) on HeLa cells are shown. DETAILED DESCRIPTION

[0085] The technical solutions of the present application will be further described in detail below in combination with specific examples. It should be understood that the following examples are only illustratively described and explained, and should not be interpreted as a limitation on the scope of protection of the present application. Any technology achieved based on the above content of the present application is covered within the scope of protection intended by the present application.

[0086] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0087] ​In the present application, the experimental methods are all conventional methods unless otherwise specified. The raw materials used are all available from public commercial channels unless otherwise specified; the percentages are all mass percentages unless otherwise specified; and the M is all mol / L unless otherwise specified.

[0088] The raw materials used in the present application are hypusine HA, hypocrellin B HB, and deacetyl hypusine HC. All the hypusine derivatives involved in the present application are shown in Examples 1-38, and the characterization results of some of the derivatives are shown in Table 1.

[0089] It should be noted that the hypusine derivatives involved in the present application that need to be protected all contain two enol tautomers, and the chemical structures of the two isomers are shown in formula (I) and (I'), formula (II) and (II'), and formula (III) and (III'), which are of course within the scope of protection. For the sake of simplicity, only one enol tautomer is listed in all the examples of the present application, and the other enol tautomer and its corresponding general structure are described in detail in the specification, and the structure is of course within the scope of protection. In addition, the general structure of the hypusine derivatives involved in the present application contains a polyethylene glycol unit (PEGn), and the number n of the unit is any integer between 0 and 100, and the corresponding chemical structure is of course within the scope of protection. For the sake of simplicity, only some integers are listed in all the examples of the present application, and the corresponding general structure of the remaining part is described in detail in the specification, and the structure is of course within the scope of protection. Any range described in the present application includes the end values and any numerical value between the end values, as well as any sub-range formed by the end values or any numerical value between the end values.

[0090] Example 1

[0091] The reaction of hypocrellin B (HB) or deacetyl hypusine (HC) with aminoethanol, aminoethanol methyl ether, and aminoethanol-polyethylene glycol, respectively, gives the 2-substituted products HB-1, HB-1-CH3, and HB-1-PEGn, and HC-1, HC-1-CH3, and HC-1-PEGn (n is an integer between 1 and 100), respectively, and their structural formulas are shown below:

[0092]

[0093] Example 2

[0094] HB (or HC) reacts with aminopropanol, aminopropanol methyl ether, aminopropanol-polyethylene glycol respectively, the 2-position substituted products of HB are HB-2, HB-2-CH3, HB-2-PEGn respectively, the 2-position substituted products of HC are HC-2, HC-2-CH3, HC-2-PEGn respectively (n is an integer between 1 and 100), the structural formulae are as shown below:

[0095]

[0096] Example 3

[0097] HB (or HC) reacts with aminopropanol, aminopropanol methyl ether, aminopropanol-polyethylene glycol respectively, the 2-position substituted products of HB are HB-2, HB-2-CH3, HB-2-PEGn respectively, the 2-position substituted products of HC are HC-2, HC-2-CH3, HC-2-PEGn respectively (n is an integer between 1 and 100), the structural formulae are as shown below:

[0098]

[0099] Example 4

[0100] HB (or HC) reacts with aminopropanol, aminopropanol methyl ether, aminopropanol-polyethylene glycol respectively, the 2-position substituted products of HB are HB-2, HB-2-CH3, HB-2-PEGn respectively, the 2-position substituted products of HC are HC-2, HC-2-CH3, HC-2-PEGn respectively (n is an integer between 1 and 100), the structural formulae are as shown below:

[0101]

[0102] Example 5

[0103] HB (or HC) reacts with aminopropanol, aminopropanol methyl ether, aminopropanol-polyethylene glycol respectively, the 2-position substituted products of HB are HB-2, HB-2-CH3, HB-2-PEGn respectively, the 2-position substituted products of HC are HC-2, HC-2-CH3, HC-2-PEGn respectively (n is an integer between 1 and 100), the structural formulae are as shown below:

[0104]

[0105] Example 6

[0106] HB (or HC) is reacted with amino polyethylene glycol, amino polyethylene glycol methyl ether, respectively, the 2-position substitution products of HB are HB-6-PEGn, HB-6-PEGn-CH3, respectively, the 2-position substitution products of HC are HC-6-PEGn, HC-6-PEGn-CH3, respectively (n is an integer between 1 and 100), the structural formulas of which are shown below:

[0107]

[0108] Example 7

[0109] HB (or HC) is reacted with amino polyethylene glycol, amino polyethylene glycol methyl ether, respectively, the 2-position substitution products of HB are HB-6-PEGn, HB-6-PEGn-CH3, respectively, the 2-position substitution products of HC are HC-6-PEGn, HC-6-PEGn-CH3, respectively (n is an integer between 1 and 100), the structural formulas of which are shown below:

[0110]

[0111] Example 8

[0112] HB (or HC) is reacted with amino polyethylene glycol, amino polyethylene glycol methyl ether, respectively, the 2-position substitution products of HB are HB-6-PEGn, HB-6-PEGn-CH3, respectively, the 2-position substitution products of HC are HC-6-PEGn, HC-6-PEGn-CH3, respectively (n is an integer between 1 and 100), the structural formulas of which are shown below:

[0113]

[0114] Example 9

[0115] HB (or HC) is reacted with amino polyethylene glycol, amino polyethylene glycol methyl ether, respectively, the 2-position substitution products of HB are HB-6-PEGn, HB-6-PEGn-CH3, respectively, the 2-position substitution products of HC are HC-6-PEGn, HC-6-PEGn-CH3, respectively (n is an integer between 1 and 100), the structural formulas of which are shown below:

[0116]

[0117] Example 10

[0118] HB (or HC) reacts with amino valeric acid, amino valeric acid methyl ester, amino valeric acid-polyethylene glycol respectively, the 2-position substituted product of HB is HB-10, HB-10-CH3, HB-10-PEGn respectively, the 2-position substituted product of HC is HC-10, HC-10-CH3, HC-10-PEGn respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0119]

[0120] Example 11

[0121] HB (or HC) reacts with amino valeric acid, amino valeric acid methyl ester, amino valeric acid-polyethylene glycol respectively, the 2-position substituted product of HB is HB-10, HB-10-CH3, HB-10-PEGn respectively, the 2-position substituted product of HC is HC-10, HC-10-CH3, HC-10-PEGn respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0122]

[0123] Example 12

[0124] HB (or HC) reacts with amino valeric acid, amino valeric acid methyl ester, amino valeric acid-polyethylene glycol respectively, the 2-position substituted product of HB is HB-10, HB-10-CH3, HB-10-PEGn respectively, the 2-position substituted product of HC is HC-10, HC-10-CH3, HC-10-PEGn respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0125]

[0126] Example 13

[0127] HB (or HC) reacts with amino valeric acid, amino valeric acid methyl ester, amino valeric acid-polyethylene glycol respectively, the 2-position substituted product of HB is HB-10, HB-10-CH3, HB-10-PEGn respectively, the 2-position substituted product of HC is HC-10, HC-10-CH3, HC-10-PEGn respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0128]

[0129] Example 14

[0130] HB (or HC) is reacted with aminomethyl sulfonic acid-polyethylene glycol, aminoethyl sulfonic acid-polyethylene glycol, respectively, the 2-position substituted product of HB is HB-16-PEGn, HB-17-PEGn, respectively, the 2-position substituted product of HC is HC-16-PEGn, HC-17-PEGn, respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0131]

[0132] Example 15

[0133] HB (or HC) is reacted with aminomethyl sulfonic acid-polyethylene glycol, aminoethyl sulfonic acid-polyethylene glycol, respectively, the 2-position substituted product of HB is HB-16-PEGn, HB-17-PEGn, respectively, the 2-position substituted product of HC is HC-16-PEGn, HC-17-PEGn, respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0134]

[0135] Example 16

[0136] HB (or HC) is reacted with aminomethyl sulfonic acid-polyethylene glycol, aminoethyl sulfonic acid-polyethylene glycol, respectively, the 2-position substituted product of HB is HB-16-PEGn, HB-17-PEGn, respectively, the 2-position substituted product of HC is HC-16-PEGn, HC-17-PEGn, respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0137]

[0138] Example 17

[0139] HB (or HC) is reacted with aminomethyl sulfonic acid-polyethylene glycol, aminoethyl sulfonic acid-polyethylene glycol, respectively, the 2-position substituted product of HB is HB-16-PEGn, HB-17-PEGn, respectively, the 2-position substituted product of HC is HC-16-PEGn, HC-17-PEGn, respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0140]

[0141] Example 18

[0142] HB (or HC) is reacted with aminomethyl sulfonic acid-polyethylene glycol, aminoethyl sulfonic acid-polyethylene glycol, respectively, the 2-position substituted product of HB is HB-16-PEGn, HB-17-PEGn, respectively, the 2-position substituted product of HC is HC-16-PEGn, HC-17-PEGn, respectively (n is an integer between 1 and 100), the structural formula is as shown below:

[0143]

[0144] Example 19

[0145] HB (or HC) respectively reacts with alkylamine of different chain length (ethylamine, propylamine, butylamine, hexylamine, octylamine), the 2-position substituted products of HB are HB-26~HB-30 respectively, and the 2-position substituted products of HC are HC-26~HC-30 respectively, the structural formulae of which are shown as follows:

[0146]

[0147] Example 20

[0148] HB (or HC) respectively reacts with hydrazine, hydroxylamine, cyclopropylamine, cyclobutylamine, cyclopentylamine, cyclohexylamine, the 2-position substituted products of HB are HB-31~HB-36 respectively, and the 2-position substituted products of HC are HC-31~HC-36 respectively, the structural formulae of which are shown as follows:

[0149]

[0150] Example 21

[0151] HB (or HC) respectively reacts with propylene amine, butylene amine, hexylene amine, octylene amine, the 2-position substituted products of HB are HB-37~HB-40 respectively, and the 2-position substituted products of HC are HC-37~HC-40 respectively, the structural formulae of which are shown as follows:

[0152]

[0153] Example 22

[0154] HB (or HC) respectively reacts with benzylamine, aminomethylpyridine, aminobutylpyridine, aminobutylpyridine salt, the 2-position substituted products of HB are HB-41~HB-44 respectively, and the 2-position substituted products of HC are HC-41~HC-44 respectively, the structural formulae of which are shown as follows:

[0155]

[0156] Example 23

[0157] HB (or HC) respectively reacts with p-, m-, o-aminomethylcyclohexanoic acid, the 2-position substituted products of HB are HB-45~HB-48 respectively, and the 2-position substituted products of HC are HC-45~HC-48 respectively, the structural formulae of which are shown as follows:

[0158]

[0159] Example 24

[0160] HB (or HC) reacted with p-, m-, o-aminomethylcyclohexanoic acid methyl ester, respectively, the 2-substituted products of HB are HB-45-CH3~HB-48-CH3, respectively, and the 2-substituted products of HC are HC-45-CH3~HC-48-CH3, respectively, whose structural formulas are shown as follows:

[0161]

[0162] Example 25

[0163] HB (or HC) reacted with p-, m-, o-aminomethylcyclohexanol, respectively, the 2-substituted products of HB are HB-49~HB-52, respectively, and the 2-substituted products of HC are HC-49~HC-52, respectively, whose structural formulas are shown as follows:

[0164]

[0165] Example 26

[0166] HB (or HC) reacted with different chain length of amino quaternary ammonium salt (counterion is Br - or I - ), respectively, the 2-substituted products of HB are HB-53~HB-56, respectively, and the 2-substituted products of HC are HC-53~HC-56, respectively, whose structural formulas are shown as follows:

[0167]

[0168] Example 27

[0169] HB (or HC) reacted with aminoethanol-different chain length quaternary ammonium salt, aminopropanol-different chain length quaternary ammonium salt (counterion is Br - or I - ), respectively, the 2-substituted products of HB are HB-57~HB-60, respectively, and the 2-substituted products of HC are HC-57~HC-60, respectively, whose structural formulas are shown as follows:

[0170]

[0171]

[0172] Example 28

[0173] HB (or HC) reacted with amino propionic acid-quaternary ammonium salt, amino butyric acid-quaternary ammonium salt (counterion is Br - or I - ), respectively, the 2-substituted products of HB are HB-61~HB-64, respectively, and the 2-substituted products of HC are HC-61~HC-64, respectively, whose structural formulas are shown as follows:

[0174]

[0175] Example 29

[0176] HB (or HC) respectively reacts with amino propionic acid-amino quaternary ammonium salt, amino butyric acid-amino quaternary ammonium salt (counter ion is Br - or I - ), the 2-position substitution products of HB are HB-65~HB-68 respectively, and the 2-position substitution products of HC are HC-65~HC-68 respectively, the structural formulae of which are shown as follows:

[0177]

[0178] Example 30

[0179] HB (or HC) respectively reacts with amino piperidine, amino morpholine, amino piperidine, the 2-position substitution products of HB are HB-69~HB-72 respectively, and the 2-position substitution products of HC are HC-69~HC-72 respectively, the structural formulae of which are shown as follows:

[0180]

[0181] Example 31

[0182] HB (or HC) respectively reacts with ethylenediamine, methyl ethylenediamine, dimethyl ethylenediamine, the 2-position substitution products of HB are HB-73~HB-76 respectively, and the 2-position substitution products of HC are HC-73~HC-76 respectively, the structural formulae of which are shown as follows:

[0183]

[0184] Example 32

[0185] HB (or HC) respectively reacts with dimethyl ethylenediamine, butyl ethylenediamine, cyclohexanediamine, the 2-position substitution products of HB are HB-77~HB-80 respectively, and the 2-position substitution products of HC are HC-77~HC-80 respectively, the structural formulae of which are shown as follows:

[0186]

[0187] Example 33

[0188] HB (or HC) is reacted with hydroxyethyl ethylenediamine, hydroxyethyl ethylenediamine-polyethylene glycol, respectively, the 2-position substituted products of HB are HB-81, HB-81-PEGn, HB-82, HB-82-PEGn, respectively, and the 2-position substituted products of HC are HC-81, HC-81-PEGn, HC-82, HC-82-PEGn, respectively (n is an integer between 1 and 100), and the structural formulas are as shown below:

[0189]

[0190] Example 34

[0191] HB (or HC) is reacted with hydroxybutyl ethylenediamine, hydroxybutyl ethylenediamine-polyethylene glycol, respectively, the 2-position substituted products of HB are HB-83, HB-83-PEGn, HB-84, HB-84-PEGn, respectively, and the 2-position substituted products of HC are HC-83, HC-83-PEGn, HC-84, HC-84-PEGn, respectively (n is an integer between 1 and 100), and the structural formulas are as shown below:

[0192]

[0193] Example 35

[0194] HB (or HC) is reacted with dihydroxyethyl ethylenediamine, trihydroxyethyl ethylenediamine-polyethylene glycol, respectively, the 2-position substituted products of HB are HB-85 to HB-88, respectively, and the 2-position substituted products of HC are HC-85 to HC-88, respectively, and the structural formulas are as shown below:

[0195]

[0196] Example 36

[0197] HB (or HC) is reacted with hydroxyethyl ethylenediamine, hydroxyethyl ethylenediamine-polyethylene glycol, respectively, the 2-position substituted products of HB are HB-81, HB-81-PEGn, HB-82, HB-82-PEGn, respectively, and the 2-position substituted products of HC are HC-81, HC-81-PEGn, HC-82, HC-82-PEGn, respectively (n is an integer between 1 and 100), and the structural formulas are as shown below:

[0198]

[0199] Example 37

[0200] HB (or HC) is reacted with hydroxypropyl ethylenediamine, hydroxypropyl ethylenediamine methyl ether and hydroxypropyl ethylenediamine-polyethylene glycol respectively, and the 2-position substitution products of HB are HB-90, HB-90-CH3 and HB-90-PEGn respectively, and the 2-position substitution products of HC are HC-90, HC-90-CH3 and HC-90-PEGn respectively, and the structural formulae are as shown below:

[0201]

[0202] Example 38

[0203] HB (or HC) is reacted with hydroxybutyl ethylenediamine, hydroxybutyl ethylenediamine methyl ether and hydroxybutyl ethylenediamine-polyethylene glycol respectively, and the 2-position substitution products of HB are HB-91, HB-91-CH3 and HB-91-PEGn respectively, and the 2-position substitution products of HC are HC-91, HC-91-CH3 and HC-91-PEGn respectively, and the structural formulae are as shown below:

[0204]

[0205] Example 39

[0206] The structural general formula of the porphyra derivative involved in the application is as shown in Figure 1 HB (or deacetyl porphyra HC) is reacted with an amino derivative, and mainly generates a porphyra 2-position amino substitution derivative as shown in formula (I); porphyra is reacted with an ethylenediamine derivative, and a porphyra ethylenediamine substitution derivative can be obtained as shown in formula (II) and formula (III). Taking the reaction of HB and amino pentanol as an example, mainly a 2-position amino substitution derivative HB-4 is generated, and the obtained product is further esterified with carboxyl polyethylene glycol to obtain HB-4-PEGn, and the synthesis method is as shown in Figure 2 Taking the reaction of HC and methyl ethylenediamine as an example, mainly HC-74 and HC-75 are generated, and the synthesis method is as shown in Figure 3 .

[0207] The derivatives shown in formula (I), (II) and (III) have a maximum absorption wavelength of about 580-650 nm, a molar extinction coefficient of 15000-40000 M -1 cm -1 , and have a strong light absorption capacity in a phototherapy window; under photosensitive conditions, not only singlet oxygen can be efficiently generated, but also a small amount of superoxide radicals can be generated. Such derivatives have good light stability and pH stability, and the specific detection results are as follows:

[0208] 1) Absorption spectrum

[0209] Figure 4 The absorption spectrum of the compound related to the present application is shown. Figure 4 In (a), the commercial porphyrin photosensitizer PpIX is multi-band absorption, all of which are narrow absorption bands, and the maximum absorption wavelength for phototherapy is 570 nm, and the molar extinction coefficient is lower 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 , and it is also narrow absorption in the phototherapy window, so the light absorption capacity of commercial PpIX and Ce6 in the phototherapy window is limited. The absorption spectrum of the derivative in the present application is completely different. As shown in Figure 4 (b), the maximum absorption peak of HB is around 470 nm, and the derivative HB-6-PEG1 in Example 6 of the present application has a wide and strong absorption in the phototherapy window, and has a very wide absorption band between 500-750 nm, and the maximum absorption peak is around 580 nm, which is red-shifted by about 110 nm from the maximum absorption peak of HB, and the molar extinction coefficient is about 20000 M -1 cm -1 , showing strong red light absorption capacity. Similarly, HC-74 is the reaction product of deacetylbaccatin in Example 31 of the present application and 2-methyl ethylenediamine, and its absorption spectrum has a greater degree of red shift, which is in the ideal phototherapy window, and has a very wide absorption band between 500-750 nm, and the main absorption peak is at 650 nm, showing strong red light absorption capacity. Other derivatives provided by the present application have similar absorption spectra to HB-6-PEG1 and HC-74, and have a very wide absorption band between 500-750 nm, and the maximum absorption peak is at 580-650 nm. Therefore, the bambusin derivatives disclosed in the present application have better light absorption capacity in the phototherapy window than the commercial photosensitizer PpIX and Ce6, and show more prominent red light absorption capacity.

[0210] 2) Reactive oxygen species

[0211] Figure 5 The reactive oxygen species of bambusin derivatives HB-45 and HC-45 (Example 23) related to the present application are tested by electron spin resonance (ESR). As shown in Figure 5 , HB-45 and HC-45 can efficiently produce reactive oxygen (ROS), which is measured by singlet oxygen and superoxide radical trapping agents, respectively. Such derivatives can efficiently produce photosensitive active species, mainly singlet oxygen, and also produce a small amount of superoxide radicals. Both of these reactive oxygen species are beneficial to photodynamic therapy. The derivatives disclosed in other examples of the present application also have the ability to efficiently produce singlet oxygen and produce a small amount of superoxide radicals.

[0212] 3) Singlet Oxygen Efficiency

[0213] The singlet oxygen efficiency of the compounds of the present application in solution was determined by the DHPA method. Figure 6 (a), 6(b) and 6(c) show the photodegradation curves of HB-73, HC-73 (Example 31) and HC-80 (Example 32), respectively. It can be seen that the photosensitizers HB-73, HC-73, HC-80 generate singlet oxygen significantly under light conditions, which in turn significantly degrade DHPA. By comparison and calculation with the singlet oxygen efficiency curve of the reference rose Bengal RB, the singlet oxygen efficiency of HB-73 and HC-73 is 0.38 and 0.40, respectively, while HC-80 generates the highest singlet oxygen efficiency of 0.45. The singlet oxygen efficiency of other derivatives of the present application is between 0.15 and 0.45, all of which have the ability to generate active oxygen efficiently.

[0214] 4) Water Solubility

[0215] Most of the derivatives disclosed in the present application contain hydrophilic groups such as polyethylene glycol, quaternary ammonium salt, sulfonic acid, carboxylic acid, etc., so that the photosensitizer molecules have strong water solubility under physiological conditions. Each milliliter of physiological saline or glucose injection can better dissolve such photosensitizer molecules, so that the photosensitizer drug can be transported well in the blood vessels without causing blood vessel blockage when intravenously injected. For example, HB-1-PEG8 (Example 1) contains 8 ethylene glycol units, and more than 5 mg can be dissolved per milliliter of physiological saline; HB-1-PEG16 contains 16 ethylene glycol units, and more than 10 mg can be dissolved per milliliter of physiological saline; HC-3-PEG6 in Example 3 can dissolve more than 5 mg per milliliter of physiological saline; HB-6-PEG16 in Example 6 can dissolve more than 15 mg per milliliter of physiological saline; HC-8-PEG12 in Example 8 can dissolve more than 10 mg per milliliter of physiological saline; these photosensitizer molecules all show excellent water solubility. Other derivatives of the present application containing strong water-soluble groups also have good water solubility and biocompatibility, and can dissolve more than 1-20 mg of photosensitizer drug molecules per milliliter of physiological saline.

[0216] 5) Photostability

[0217] The photostability of the derivatives of the present application and commercial photosensitizers is compared as shown in Figure 7 It can be seen that the 635 nm laser at 20 mW / cm 2After 30 minutes of illumination at a given light intensity, the absorption spectra of HB-2-PEG4 and HC-2-PEG8 (Example 2) did not show a significant decrease, with the absorption intensity at the maximum wavelength decreasing by less than 10%, indicating good photostability. The derivatives HC-73 (Example 31) and HC-80 (Example 32) also showed a decrease in absorption intensity at the maximum wavelength of less than 10%, similarly exhibiting good photostability. Under the same conditions, a 635nm laser at 20mW / cm² was used. 2 After 30 minutes of illumination under light intensity, the maximum absorption of commercial photosensitizers Ce6 and HpD decreased by 30% and 50%, respectively. Other derivatives of this invention also exhibit good photostability, with their maximum wavelength absorption intensity decreasing by less than 10% under the same conditions. Therefore, the bamboo red mycotoxin derivatives of this invention have better photostability than commercial photosensitizers.

[0218] 6) pH stability

[0219] The pH stability of the bamboo red mycotoxin derivative of the present invention is as follows: Figure 8 As shown in the figure, the absorption spectra of these derivatives do not change significantly within the pH range of 6.2–8.0, indicating that they exhibit good pH stability under physiological conditions. As shown in the figure, HB-3-PEG2 and HC-3-PEG8 (Example 3) show no significant change in absorption spectra within the pH range of 6.2–8.0; HC-73 (Example 31) and HB-77 (Example 32) also show no significant change in absorption spectra within the pH range of 6.2–8.0. This good pH stability within this range is because the two phenolic hydroxyl groups of porphyrin are less prone to deprotonation under these conditions. In contrast, commercially available hematoporphyrin HpD contains two carboxyl groups, which can be deprotonated within the pH range of 6.2–8.0, resulting in a significant change in the absorption spectrum and thus demonstrating the instability of the HpD photosensitizer. Other derivatives of this invention also exhibit good pH stability under physiological conditions, with minimal impact on the absorption spectrum of the photosensitizing drugs from pH variations within the 6.2–8.0 range. Therefore, the bamboo red mycotoxin derivative of the present invention has better pH stability than commercial porphyrin HpD.

[0220] The structures of some bamboo red fungicide derivatives involved in this invention are shown in Examples 1-38. Some bamboo red fungicide derivatives with 2-amino or ethylenediamine substitution have a maximum absorption wavelength of approximately 580-630 nm and a molar extinction coefficient reaching 15000-40000 MHz. -1 cm -1 It has a strong light absorption capacity during phototherapy; under photosensitivity conditions, it can efficiently generate reactive oxygen species such as singlet oxygen, with singlet oxygen efficiency reaching up to about 40%; these derivatives have good photostability and pH stability, and some of their photophysical data are shown in Table 1.

[0221] Table 1: Photophysical data of some bamboo red fungin derivatives of the present invention

[0222]

[0223] Example 40

[0224] Tumor cell culture: Various cell lines (esophageal cancer cells AKR, gastric cancer cells MFC, lung cancer cells A549, liver cancer cells HepG2, bile duct cancer cells MCC, colon cancer cells HCT116, brain cancer cells G442, head and neck cancer cells SCC2, tongue cancer cells TSCCa, nasal cancer cells KB, oral cancer cells CAL27, glioma cells C6, basal cell carcinoma cells BCC, squamous cell carcinoma cells PECA, melanoma cells B16, cutaneous T-cell lymphoma cells HH, prostate cancer cells LNCaP, and bladder cancer cells MBT-2) were provided by the Cell Center of Peking Union Medical College. The culture conditions for the above cells were: RPMI-1640 medium supplemented with 10% FBS, 1% streptomycin (100 μg / mL), and penicillin (100 μg / mL), and cultured in a 37℃, 5% CO2 incubator. Tumor cells were seeded in glass-based confocal dishes, and a solution of 100 μL of the photosensitizing agent erythromycin was added to the culture medium, and the dishes were incubated for 4 hours. The cells were carefully washed twice with pre-cooled PBS solution to remove photosensitizing drugs that had not entered the cells. The cultured tumor cells were used for MTT assays.

[0225] Example 41

[0226] The cell toxicity assay, using HB-1-PEG8 (Example 1) as an example, was performed as follows: Cultured lung cancer cells (A549 cells) were digested with 0.25% trypsin to prepare a single-cell suspension, which was then seeded in 96-well culture plates and incubated. After cell attachment, the supernatant was discarded, and different concentrations of photosensitizers (such as HB-1-PEG8) were added under dark conditions for 1 hour of incubation. Cell viability was then assessed using the MTT assay. Alternatively, 20 μL of MTT was added to each well, and the cells were incubated for 4 hours. The supernatant was discarded, and 150 μL of DMSO was added to each well. The plates were shaken for 10 minutes with a micro-shaker to fully dissolve the purple crystals. The OD value (570 nm) of each well was measured using a microplate reader, and cell viability was calculated. Cell viability = (OD value of experimental group / OD value of blank group) × 100%. Figure 10-2As shown, cytotoxicity (dark toxicity) studies indicate that HB-1-PEG8 exhibits low cytotoxicity, similar to the commercial photosensitizer HpD. Incubation with a 20 μM concentration of the photosensitizer for half an hour did not result in significant A549 cell death, indicating that this type of photosensitizer is essentially non-cytotoxic. The dark toxicity of other derivatives of this invention is similar to that of HB-1-PEG8, as shown in Figures 10-14 and Tables 2-4. These derivatives are essentially non-cytotoxic, with cell viability exceeding 90% at concentrations up to 20 μM.

[0227] Example 42

[0228] Cell phototoxicity experiments, using HC-1-PEG8 synthesized in Example 1 as an example:

[0229] The experimental procedure was the same as the dark toxicity experiment, using a 635nm semiconductor laser (20mW / cm²). 2 Irradiate the plate so that the beam is evenly and perpendicularly directed onto the 96-well culture plate for 1000 seconds. Figure 10-1 The phototoxicity experiments shown demonstrate that HC-1-PEG8 exhibits a very strong killing effect on A549 cells under red light irradiation. A concentration of 50 nM can kill more than 90% of A549 cells, while commercial photosensitizers under the same conditions can only kill about 20% of A549 cells. This indicates that the photodynamic effect of this derivative is significantly superior to the commercial photosensitizer HpD. The phototoxicity of other derivatives of this invention is similar to that of HC-1-PEG8, and the results are shown in Figures 10-14. The synthesized derivatives can kill more than 80-90% of tumor cells at a concentration of 50 nM, with a median lethal concentration (IC50) of 1 / 3. 50 The value is approximately 20–30 nM. Therefore, the bamboo red mycotoxin derivative disclosed in this invention has better photodynamic effects than the commercial photosensitizer HpD.

[0230] Example 43

[0231] Animal experiments: All animal experimental procedures complied with the regulations concerning animal use and husbandry stipulated by the Laboratory Animal Use and Management Committee of the Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences. Tumor models were established using 4-6 week old female Balb / c nude mice (approximately 20g). 50μL of suspensions of various tumor cells (5×10⁻⁶) were injected into the right posterior thigh of each mouse. 6 (Number of tumors). The tumor volume grew to approximately 200 mm. 3 At that time, in vivo fluorescence imaging and photodynamic therapy experiments were conducted.

[0232] Small animal fluorescence imaging experiment: Tumor-bearing mice were injected intravenously with 40 μL of a photosensitizing drug in physiological saline solution (10 mg / mL) at a mouse dose (10 mg / kg). Tumor fluorescence images were then collected at 0, 1, 2, 4, 6, and 8 hours using in vivo small animal imaging. After 8 hours, the tumor-bearing mice were euthanized by cervical dislocation, and the tumors and major organs were removed for in vitro fluorescence imaging. The average fluorescence intensity within the ROS region was statistically analyzed for semi-quantitative analysis.

[0233] Photodynamic therapy experiment: When the tumor volume reaches 200mm 3 At that time, tumor-bearing mice were randomly divided into 3 groups (n=5 per group): no treatment (Group I); only tail vein injection of photosensitizing drug (100 μL, 500 μM) (Group II); and tail vein injection of photosensitizing drug (100 μL, 500 μM) followed by 635 nm laser irradiation of the tumor site (10 min, 0.1 W cm⁻¹) 4 h later. -2 (Group III). Tumor volume and body weight were measured every other day after laser treatment, and survival rate was recorded.

[0234] Example 44

[0235] As described above, the pharmacological activity of the bamboo red fungicide derivative of the present invention was tested, and the specific results are as follows:

[0236] 1) Confocal fluorescence imaging of bamboo red fungus derivatives in cancer cells

[0237] Cell fluorescence imaging of the bamboo red fungin derivative of the present invention, as shown in the figure Figure 9 As shown, the photosensitizer HB-1-PEG12 exhibits good water solubility and biocompatibility. Co-incubation with lung cancer cells (A549) revealed that the photosensitizer rapidly entered the lysosomes of the cells and produced red fluorescence, indicating that HB-1-PEG12 can be used for fluorescence imaging of lung cancer cells. Fluorescence detection can be used to track the accumulation, distribution, and metabolism of the drug in vivo. Using DCFH-DA to detect intracellular singlet oxygen, co-incubation of the photosensitizer HB-1-PEG12 and the fluorescent probe DCFH-DA in cells showed that the green fluorescence gradually increased with irradiation time up to 120 s, indicating an increase in intracellular singlet oxygen. Other derivatives of this invention also effectively entered the lysosomes of lung cancer cells and were used for cell fluorescence imaging, allowing for the tracking of drug accumulation, distribution, and metabolism in vivo.

[0238] 2) Dark toxicity and phototoxicity tests of bamboo red fungicide derivatives on gastrointestinal tumor cells

[0239] Digestive tract tumor cells mainly include esophageal cancer cells, gastric cancer cells, lung cancer cells, liver cancer cells, bile duct cancer cells, and colon cancer cells.

[0240] The derivatives of the present application were incubated with esophageal cancer cells AKR cells, as shown in Figure 10-1 (a), the cell dark toxicity study of the photosensitizer showed that the cell toxicity of HB-1-PEG4 and HC-1-PEG8 was small, similar to the commercial photosensitizer HpD, whether there were 4 PEG units or 8 PEG units. There was no obvious death of esophageal cancer cells after incubation with 20 μM photosensitizer HB-1-PEG4 or HC-1-PEG8 for half an hour, indicating that the photosensitizer had no cell toxicity. As shown in Figure 10-1 (b), the cell phototoxicity study showed that the photosensitizer had very strong killing effect on esophageal cancer cells under red light irradiation (10 min, 0.1 W cm -2 ). More than 90% of esophageal cancer cells were killed at a concentration of 50 nM HB-1-PEG4, and the half lethal concentration IC 50 was about 25 nM; more than 90% of esophageal cancer cells were killed at a concentration of 50 nM HC-1-PEG8, and the half lethal concentration IC 50 was about 25 nM; and the commercial photosensitizer HpD could only kill about 30% of esophageal cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives on esophageal cancer was significantly better than that of the commercial photosensitizer.

[0241] The derivatives of the present application were incubated with lung cancer cells A549, as shown in Figure 10-2 (a), the cell dark toxicity study showed that the cell toxicity of HB-3-PEG8 and HC-3-PEG16 (Example 3) was small, similar to the commercial photosensitizer blood porphyrin HpD. There was no obvious death of lung cancer cells after incubation with 20 μM photosensitizer HB-3-PEG8 or HC-3-PEG16 for half an hour, indicating that the photosensitizer had no cell toxicity. As shown in Figure 10-2 (b), the cell phototoxicity experiment showed that the photosensitizer had very strong killing effect on lung cancer cells under red light irradiation. More than 90% of lung cancer cells were killed at a concentration of 50 nM HB-3-PEG8 or HC-3-PEG16, and the half lethal concentration IC 50 was about 30 nM; and the commercial photosensitizer HpD could only kill about 20% of lung cancer cells under the same conditions, indicating that the photodynamic effect of the phytomenadione derivatives was significantly better than that of the commercial photosensitizer.

[0242] The derivatives of the present application were incubated with liver cancer cells HepG2, as shown in Figure 10-3(a) shows, the dark toxicity study of photosensitizer shows that the cytotoxicity of HB-6-PEG2 and HC-6-PEG6 prepared in Example 6 is small, similar to the commercial photosensitizer HpD. The hepatoma cells are incubated with 20 μM concentration of photosensitizer for half an hour, and no obvious death of hepatoma cells is observed, indicating that the photosensitizer basically has no cytotoxicity. As shown in Figure 10-3 (b) shows the cell phototoxicity study, which shows that the photosensitizer exhibits very strong killing effect on hepatoma cells under red light irradiation. Both HB-6-PEG2 and HC-6-PEG12 at 50 nM concentration can kill more than 85% of hepatoma cells, and the half lethal concentration IC 50 is about 30 nM; while the commercial hematoporphytin derivative HpD can only kill about 20% of hepatoma cells under the same conditions, indicating that the photodynamic effect of the bamboo red bacterium derivative is obviously better than the commercial photosensitizer.

[0243] The derivative in the application is incubated with colon cancer cells HCT116, as shown in Figure 10-4 (a) shows the dark toxicity study of photosensitizer, which shows that the cytotoxicity of HB-14-PEG6 or HC-14-PEG12 prepared in Example 13 is small, similar to the commercial photosensitizer HpD. The colon cancer cells are incubated with 20 μM concentration of photosensitizer for half an hour, and no obvious death of colon cancer cells is observed, indicating that the photosensitizer basically has no cytotoxicity. As shown in Figure 10-4 (b) shows the cell phototoxicity study, which shows that the photosensitizer exhibits very strong killing effect on colon cancer cells under red light irradiation. Both HB-14-PEG6 and HC-14-PEG12 at 50 nM concentration can kill more than 85% of colon cancer cells, and the half lethal concentration IC 50 is about 25 nM; while the commercial photosensitizer HpD can only kill about 20% of colon cancer cells under the same conditions, indicating that the photodynamic effect of the derivative is obviously better than the commercial photosensitizer HpD.

[0244] The derivative in the application is incubated with cholangiocarcinoma cells MCC, as shown in Figure 10-5 (a) shows the dark toxicity study of photosensitizer, which shows that the cytotoxicity of HB-19-PEG4 and HC-19-PEG8 prepared in Example 15 is small, similar to the commercial photosensitizer HpD. The cholangiocarcinoma cells are incubated with 20 μM concentration of photosensitizer for half an hour, and no obvious death of cholangiocarcinoma cells is observed, indicating that the photosensitizer basically has no cytotoxicity. As shown in Figure 10-5 (b) shows the cell phototoxicity study, which shows that the photosensitizer exhibits very strong killing effect on cholangiocarcinoma cells under red light irradiation. Both HB-19-PEG4 and HC-19-PEG8 at 50 nM concentration can kill more than 85% of cholangiocarcinoma cells, and the half lethal concentration IC 50The IC50value is about 30 nM; while the commercial photosensitizer HpD can only kill about 20% of the cholangiocarcinoma cells under the same conditions, indicating that the photodynamic effect of the bamboo red bacterium derivatives is obviously better than that of the commercial photosensitizer HpD.

[0245] The derivatives in the present application were incubated with gastric cancer cells MFC, as shown in Figure 10-6 As shown in (a), the dark toxicity study of the photosensitive drug shows that the cell toxicity of HB-45 and HC-45 prepared in Example 23 is small, similar to that of the commercial photosensitive drug hematoxylin HpD. The photosensitizer was incubated with gastric cancer cells at a concentration of 20 μM for half an hour, and no obvious death of MFC cells was observed, indicating that the photosensitizer has little cytotoxicity. As shown in Figure 10-6 (b), the cell phototoxicity study shows that the photosensitive drug exhibits very strong killing power on gastric cancer cells under red light irradiation. 50 nM concentration of HB-45 and HC-45 can kill more than 85% of gastric cancer cells, and the IC 50 The IC50value is about 30 nM; while the commercial photosensitizer HpD can only kill about 20% of the cholangiocarcinoma cells under the same conditions, indicating that the photodynamic effect of the bamboo red bacterium derivatives is obviously better than that of the commercial photosensitizer HpD.

[0246] The above detailed examples illustrate that part of the bamboo red bacterium derivatives disclosed in the present application can efficiently kill esophageal cancer AKR, gastric cancer MFC, lung cancer A549, liver cancer HepG2, cholangiocarcinoma MCC, colon cancer HCT116 and other digestive tract tumor cells. Whether it is a bamboo red bacterium (HB) or a derivative of deacetyl bamboo red bacterium (HC), the light has little damage to the cells, and under light irradiation, it has a very strong ability to inactivate the above-mentioned digestive tract tumor cells. In addition, bamboo red bacterium derivatives with different lengths of PEG chains, different lengths of sulfonic acid groups and different lengths of quaternary ammonium salt chains all exhibit good photodynamic inactivation ability to tumor cells. Therefore, such derivatives can efficiently kill esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer and other digestive tract tumor cells, and the related data are shown in Table 2.

[0247] Table 2: MTT data of part of the bamboo red bacterium derivatives of the present application on digestive tract tumor cells

[0248]

[0249]

[0250] 3) Dark toxicity and phototoxicity test of bamboo red bacterium derivatives on head and neck facial tumor cells

[0251] The head and neck facial tumor cells mainly include head and neck cancer cells, brain cancer cells, tongue cancer cells, nasal cancer cells, oral cancer cells and brain glioma cells.

[0252] The bamboo red bacterium derivative disclosed in the present application can efficiently kill brain cancer, head and neck cancer, tongue cancer, nose cancer, oral cancer, brain glioma and other head and neck facial tumor cells under 635 nm laser irradiation. The bamboo red bacterium derivative disclosed in the examples is taken as an example to illustrate the phototoxicity and dark toxicity of the bamboo red bacterium derivative on head and neck facial tumor cells.

[0253] The dark toxicity of the photosensitizer is studied by incubating the derivative in the present application with brain cancer cells G442, and it is shown that the cell toxicity of HB-10-PEG4 and HC-10-PEG8 (Example 10) is small, similar to that of the commercial photosensitizer hematoporphyrin HpD, whether there are 4 PEG units or 8 PEG units. The brain cancer cells are incubated with 20 μM concentration of the photosensitizer HB-10-PEG4 and HC-10-PEG8 for half an hour, and no obvious death of the brain cancer cells is observed, indicating that the photosensitizer has no cell toxicity. Figure 11 The cell phototoxicity study shown in (a) shows that the photosensitizer exhibits very strong killing effect on brain cancer cells under red light irradiation. The 50 nM concentration of HB-10-PEG4 and HC-10-PEG8 can kill more than 85% of the brain cancer cells, and the half lethal concentration IC 50 is about 30 nM; and the commercial photosensitizer HpD can only kill about 20% of the brain cancer cells under the same conditions, indicating that the photodynamic effect of the bamboo red bacterium derivative is obviously better than that of the commercial photosensitizer HpD.

[0254] The dark toxicity of the photosensitizer is studied by incubating the derivative in the present application with head and neck cancer cells SCC2, and it is shown that the cell toxicity of HB-12-PEG8 and HC-12-PEG12 prepared in Example 12 is small, similar to that of the commercial photosensitizer hematoporphyrin HpD. The head and neck cancer cells are incubated with 20 μM concentration of the photosensitizer for half an hour, and no obvious death of the head and neck cancer cells is observed, indicating that the photosensitizer has no cell toxicity. Figure 11 The cell phototoxicity study shown in (b) shows that the photosensitizer exhibits very strong killing effect on head and neck cancer cells under red light irradiation. The 50 nM concentration of HB-12-PEG8 and HC-12-PEG12 can kill more than 85% of the head and neck cancer cells, and the half lethal concentration IC 50 is about 30 nM; and the commercial photosensitizer HpD can only kill about 20% of the head and neck cancer cells under the same conditions, indicating that the photodynamic effect of the bamboo red bacterium derivative is obviously better than that of the commercial photosensitizer HpD.

[0255] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the present application with tongue cancer cells TSCCa. The results showed that the cell toxicity of HB-20-PEG6 and HC-20-PEG12 prepared in Example 16 was small, similar to that of the commercial photosensitizer HpD. The tongue cancer cells were incubated with 20 μM photosensitizer for half an hour, and no obvious death of the tongue cancer cells was observed, indicating that the photosensitizer had no cell toxicity. Figure 11 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on the tongue cancer cells under red light irradiation. HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM could kill more than 85% of the tongue cancer cells, and the half lethal concentration IC50was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of the tongue cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives was obviously superior to that of the commercial photosensitizer. 50 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on the tongue cancer cells under red light irradiation. HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM could kill more than 85% of the tongue cancer cells, and the half lethal concentration IC50was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of the tongue cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives was obviously superior to that of the commercial photosensitizer.

[0256] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the present application with tongue cancer cells TSCCa. The results showed that the cell toxicity of HB-20-PEG6 and HC-20-PEG12 prepared in Example 16 was small, similar to that of the commercial photosensitizer HpD. The tongue cancer cells were incubated with 20 μM photosensitizer for half an hour, and no obvious death of the tongue cancer cells was observed, indicating that the photosensitizer had no cell toxicity. Figure 12 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on the tongue cancer cells under red light irradiation. HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM could kill more than 85% of the tongue cancer cells, and the half lethal concentration IC50was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of the tongue cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives was obviously superior to that of the commercial photosensitizer. 50 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on the tongue cancer cells under red light irradiation. HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM could kill more than 85% of the tongue cancer cells, and the half lethal concentration IC50was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of the tongue cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives was obviously superior to that of the commercial photosensitizer.

[0257] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the present application with tongue cancer cells TSCCa. The results showed that the cell toxicity of HB-20-PEG6 and HC-20-PEG12 prepared in Example 16 was small, similar to that of the commercial photosensitizer HpD. The tongue cancer cells were incubated with 20 μM photosensitizer for half an hour, and no obvious death of the tongue cancer cells was observed, indicating that the photosensitizer had no cell toxicity. Figure 12 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on the tongue cancer cells under red light irradiation. HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM could kill more than 85% of the tongue cancer cells, and the half lethal concentration IC50was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of the tongue cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives was obviously superior to that of the commercial photosensitizer. 50 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on the tongue cancer cells under red light irradiation. HB-20-PEG6 and HC-20-PEG12 at a concentration of 50 nM could kill more than 85% of the tongue cancer cells, and the half lethal concentration IC50was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of the tongue cancer cells under the same conditions, indicating that the photodynamic effect of the derivatives was obviously superior to that of the commercial photosensitizer.

[0258] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the application with C6 glioma cells. The results showed that the derivatives of the application, HB-73 prepared in Example 31 and HC-80 prepared in Example 32, had less cytotoxicity to the glioma cells than the commercial photosensitizer, HpD. For example, Figure 12 The results of the cell phototoxicity study shown in Figure (c) showed that the photosensitizer had very strong killing effect on the glioma cells under red light irradiation. HB-73 and HC-80 at a concentration of 50 nM could kill more than 85% of the glioma cells, and the half lethal concentration IC50was about 30 nM. However, the commercial photosensitizer, HpD, could only kill about 20% of the C6 cells under the same conditions, which indicated that the photodynamic effect of the derivatives of the application was significantly better than that of the commercial photosensitizer, HpD. 50 The results of the cell phototoxicity study shown in Figure (c) showed that the photosensitizer had very strong killing effect on the glioma cells under red light irradiation. HB-73 and HC-80 at a concentration of 50 nM could kill more than 85% of the glioma cells, and the half lethal concentration IC50was about 30 nM. However, the commercial photosensitizer, HpD, could only kill about 20% of the C6 cells under the same conditions, which indicated that the photodynamic effect of the derivatives of the application was significantly better than that of the commercial photosensitizer, HpD.

[0259] The above detailed examples show that some of the derivatives of the application can efficiently kill head and neck tumor cells such as brain cancer G442, head and neck cancer SCC2, tongue cancer TSCCa, nose cancer KB, oral cancer CAL27, and glioma C6. Neither HB nor HC derivatives have any damage to the cells without light, but have very strong ability to inactivate the above-mentioned digestive tract tumor cells under light. In addition, the derivatives of the application with different lengths of PEG chains, different lengths of sulfonic acid chains, and different lengths of quaternary ammonium salt chains all show good photodynamic inactivation ability to tumor cells. Therefore, the derivatives of the application can efficiently kill brain cancer, head and neck cancer, tongue cancer, nose cancer, oral cancer, glioma, and other head and neck tumor cells. The related data are shown in Table 3.

[0260] Table 3: MTT data of some derivatives of the application on head and neck tumor cells

[0261]

[0262]

[0263] 4) Dark toxicity and phototoxicity test of the derivatives of the application on reproductive and urinary system tumor cells

[0264] The derivatives of the application disclosed in the application can efficiently kill reproductive and urinary system tumor cells such as basal cell carcinoma, skin T-cell lymphoma, melanoma, squamous skin cancer, prostate cancer, and bladder cancer under 635 nm laser irradiation. The phototoxicity and dark toxicity of the derivatives of the application disclosed in the examples on skin tumor and urinary system tumor cells are described.

[0265] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the present application with basal cell carcinoma cells BCC. The results showed that the cell toxicity of HB-4-PEG8 and HC-4-PEG16 (Example 4) was small, similar to that of the commercial photosensitizer hematoporphyrin HpD. No obvious death of basal cell carcinoma cells was observed after incubation with 20 μM photosensitizer for half an hour, indicating that the photosensitizer had little cell toxicity. As shown in Figure 13 The cell phototoxicity study shown in (a) indicated that the photosensitizer had very strong killing effect on basal cell carcinoma cells under red light irradiation. HB-4-PEG8 and HC-4-PEG16 at a concentration of 50 nM could kill more than 90% of basal cell carcinoma cells, and the half lethal concentration IC50 was about 30 nM; while the commercial photosensitizer hematoporphyrin derivative HpD could only kill about 20% of basal cell carcinoma cells under the same conditions, indicating that the photodynamic effect of the bambusurin polyethylene glycol derivatives was obviously superior to that of the commercial photosensitizer hematoporphyrin HpD. 50 The cell phototoxicity study shown in (a) indicated that the photosensitizer had very strong killing effect on basal cell carcinoma cells under red light irradiation. HB-4-PEG8 and HC-4-PEG16 at a concentration of 50 nM could kill more than 90% of basal cell carcinoma cells, and the half lethal concentration IC50 was about 30 nM; while the commercial photosensitizer hematoporphyrin derivative HpD could only kill about 20% of basal cell carcinoma cells under the same conditions, indicating that the photodynamic effect of the bambusurin polyethylene glycol derivatives was obviously superior to that of the commercial photosensitizer hematoporphyrin HpD.

[0266] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the present application with melanoma cells B16. The results showed that the cell toxicity of HB-9-PEG4 and HC-9-PEG8 prepared in Example 9 was small, similar to that of the commercial photosensitizer hematoporphyrin HpD. As shown in Figure 13 The cell phototoxicity study shown in (b) indicated that the photosensitizer had very strong killing effect on melanoma cells under red light irradiation. HB-9-PEG4 and HC-9-PEG8 at a concentration of 50 nM could kill more than 85% of melanoma cells, and the half lethal concentration IC50 was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of melanoma cells under the same conditions, indicating that the photodynamic effect of the bambusurin derivatives was obviously superior to that of the commercial photosensitizer HpD. 50 The cell phototoxicity study shown in (b) indicated that the photosensitizer had very strong killing effect on melanoma cells under red light irradiation. HB-9-PEG4 and HC-9-PEG8 at a concentration of 50 nM could kill more than 85% of melanoma cells, and the half lethal concentration IC50 was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of melanoma cells under the same conditions, indicating that the photodynamic effect of the bambusurin derivatives was obviously superior to that of the commercial photosensitizer HpD.

[0267] The dark toxicity of the photosensitizer was studied by incubating the derivatives of the present application with squamous skin cancer cells PECA. The results showed that the cell toxicity of HB-36 and HC-36 prepared in Example 20 was small, similar to that of the commercial photosensitizer hematoporphyrin HpD. As shown in Figure 13 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on PECA cells under red light irradiation. 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 IC50 was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of PECA cells under the same conditions, indicating that the photodynamic effect of the bambusurin derivatives was obviously superior to that of the commercial photosensitizer HpD. 50 The cell phototoxicity study shown in (c) indicated that the photosensitizer had very strong killing effect on PECA cells under red light irradiation. 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 IC50 was about 30 nM; while the commercial photosensitizer HpD could only kill about 20% of PECA cells under the same conditions, indicating that the photodynamic effect of the bambusurin derivatives was obviously superior to that of the commercial photosensitizer HpD.

[0268] The derivatives of this invention were incubated with prostate cancer cells LNCaP. A dark toxicity study of the photosensitizing drug showed that the HB-70 and HC-71 cells prepared in Example 30 exhibited low cytotoxicity, similar to the commercially available photosensitizing drug hematoporphyrin HpD. Figure 14 The phototoxicity study shown in (a) indicates that the photosensitizing drugs exhibit a very strong killing effect on prostate cancer cells under red light irradiation. Both HB-70 and HC-71 at a concentration of 50 nM can kill more than 85% of prostate cancer cells, with a median lethal concentration (IC50) of 1 / 3. 50 The value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of prostate cancer cells, indicating that the photodynamic effect of this type of bamboo red fungicide derivative is significantly better than that of the commercial photosensitizer HpD.

[0269] The derivatives of this invention were incubated together with MBT-2 bladder cancer cells. A dark cytotoxicity study of the photosensitizing drug showed that the HB-73 and HC-73 cells prepared in Example 31 exhibited low cytotoxicity, similar to the commercially available photosensitizing drug hematoporphyrin HpD. Figure 14 (b) The phototoxicity study showed that the photosensitizing drugs exhibited a very strong killing effect on bladder cancer cells under red light irradiation. Both HB-73 and HC-73 at a concentration of 50 nM could kill more than 85% of bladder cancer cells, with an IC50 (median lethal concentration) of [missing value]. 50 The value is approximately 25 nM; while under the same conditions, the commercial photosensitizer HpD can only kill about 20% of bladder cancer cells, indicating that the photodynamic effect of this type of bamboo red fungus polyglycol derivative is significantly better than that of commercial photosensitizers.

[0270] The derivatives of this invention were incubated with skin T-cell lymphoma cells HH. Dark toxicity studies of the photosensitizing drugs showed that HC-80 prepared in Example 32 and HC-81 prepared in Example 33 exhibited low cytotoxicity, similar to the commercially available photosensitizing drug hematoporphyrin HpD. Figure 14 (c) The phototoxicity study showed that the photosensitizing drugs exhibited very strong killing power against skin T-cell lymphoma cells under red light irradiation. Both HC-80 and HC-81 at a concentration of 50 nM could kill more than 85% of HH cells, with a median lethal concentration (IC50) of 100%. 50 The value is approximately 30 nM; while under the same conditions, the commercial photosensitizer HpD can only kill 20% of HH cells, indicating that the photodynamic effect of this type of bamboo red mycotoxin derivative is significantly better than that of commercial photosensitizers.

[0271] The above detailed examples show that some of the porfimer derivatives disclosed in the present application can efficiently kill basal cell carcinoma BCC, squamous cell carcinoma PECA, melanoma B16, cutaneous T-cell lymphoma HH, prostate cancer LNCaP, bladder cancer MBT-2 and other skin and urogenital tumor cells. Whether the porfimer or the derivative of deacetyl porfimer, the light has little damage to the cells, and the light has a strong ability to inactivate the above-mentioned digestive tract tumor cells. In addition, the porfimer derivatives with different PEG chain lengths, different sulfonic acid group chain lengths, and different quaternary ammonium salt chain lengths all show good photodynamic inactivation of tumor cells. Therefore, such derivatives can efficiently kill basal cell carcinoma, squamous cell carcinoma, melanoma, cutaneous T-cell lymphoma, prostate cancer, bladder cancer and other reproductive and urogenital tumor cells, and the detailed data are shown in Table 4.

[0272] Table 4: MTT data of some porfimer derivatives of the present application on skin tumor and reproductive and urogenital tumor cells

[0273]

[0274]

[0275] The above studies show that by changing the molecular structure and different PEG chain lengths and end groups, the porfimer derivatives of the present application have good photodynamic killing effect on various tumor cells: Figures 10-14 and Tables 2-4 show that such derivatives have little cytotoxicity without light, and have obvious photodynamic effect under light, and 50nM concentration of photosensitizer can efficiently kill most of various tumor cells, and the half lethal concentration IC 50 is about 20-30nM, which is 1-2 orders of magnitude lower than the IC 50 of the commercial photosensitizer HpD under the same conditions. Other derivatives of the present application also have dark toxicity and phototoxicity as shown in Figures 10-14. Therefore, the porfimer derivatives disclosed in the present application with 2-amino substitution or ethylenediamine substitution have better dark toxicity and photodynamic effect than the commercial photosensitizer HpD.

[0276] Example 45 Animal imaging

[0277] Fluorescence imaging of the foregoing cells Figure 9) show that the derivatives provided by the present application can enter tumor cells well to produce near-infrared fluorescence imaging. In order to study the enrichment and metabolism of such derivatives at the tumor in vivo, the present application uses tumor-bearing mice of various different tumor cells as models to study the enrichment process of such derivatives at the tumor in mice by small animal live fluorescence imaging. The derivatives with good water solubility such as polyethylene glycol, sulfonic acid group, carboxylic acid group, quaternary ammonium salt, etc. achieve the targeted enrichment of drugs at the tumor by tail vein injection; and for derivatives with slightly poor water solubility, intratumoral injection of tumor-bearing mice is used to achieve the enrichment of photosensitizing drugs in the tumor area.

[0278] 1) Drug enrichment in gastrointestinal tumor mice (esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, colon cancer)

[0279] The esophageal cancer AKR cells were used to inoculate subcutaneous tumors to obtain an esophageal cancer subcutaneous mouse tumor model. HB-1-PEG4 (Example 1) was used as a photosensitizing drug, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg to observe its fluorescence imaging behavior in the esophageal cancer tumor-bearing mice. The fluorescence signals at the tumor site for 4 hours were collected using a multispectral small animal live imaging system. As shown in Figure 15 (a), after several cycles in vivo by intravenous injection, the drug molecule HB-1-PEG4 has a certain drug enrichment at the tumor site by passive targeting for about 4 hours, which belongs to moderate 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 moderate enrichment at the tumor site.

[0280] The gastric cancer MFC cells were used to inoculate subcutaneous tumors to obtain a gastric cancer subcutaneous mouse tumor model. HC-3-PEG12 (Example 3) was used as a photosensitizing drug, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg to observe the fluorescence imaging behavior, and the fluorescence signals at the tumor site for 4 hours were collected. As shown in Figure 15 (b), after several cycles in vivo by intravenous injection, the drug molecule HC-3-PEG12 has very strong drug enrichment at the tumor site by passive targeting for about 4 hours. The reason is that this derivative molecule contains 12 polyethylene glycol units, making the water solubility of the drug molecule good, and the hydrophilic-hydrophobic ratio of the whole molecule appropriate, resulting in strong enrichment at the tumor site.

[0281] The lung cancer A549 cells were used to inoculate subcutaneous tumors to obtain a lung cancer subcutaneous mouse tumor model. HB-6-PEG8 (Example 6) was used as a photosensitizing drug, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg to observe its fluorescence imaging behavior in the lung cancer tumor-bearing mice, and the fluorescence signals at the tumor site for 4 hours were collected. As shown in Figure 15(c) shows that the drug molecule HB-6-PEG8 has a strong enrichment in the tumor site after several cycles in vivo by passive targeting for about 4 h after intravenous injection, because this derivative molecule contains 8 polyethylene glycol units, making the water solubility of the molecule better, and the whole molecule has a suitable hydrophilic-hydrophobic ratio, resulting in a stronger enrichment in the tumor site.

[0282] A subcutaneous tumor model of hepatocarcinoma was obtained by subcutaneous inoculation of HepG2 cells. HC-9-PEG10 (Example 9) was used as a photosensitizing drug, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-9-PEG10 in the tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 h. As shown in Figure 15 (d) shows that the drug molecule HB-7-PEG16 has a strong enrichment in the tumor site after several cycles in vivo by passive targeting for about 4 h after intravenous injection, because this derivative molecule contains 16 polyethylene glycol units, making the water solubility of the whole drug molecule better, and the whole molecule has a suitable hydrophilic-hydrophobic ratio, resulting in a stronger enrichment in the tumor site.

[0283] A subcutaneous tumor model of hepatocarcinoma was obtained by subcutaneous inoculation of HepG2 cells. HC-9-PEG10 (Example 9) was used as a photosensitizing drug, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-9-PEG10 in the tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 h. As shown in Figure 15 (e) shows that the drug molecule HC-9-PEG10 has a strong enrichment in the tumor site after several cycles in vivo by passive targeting for about 4 h after intravenous injection, because this derivative molecule contains 10 polyethylene glycol units, making the water solubility of the whole drug molecule better, and the whole molecule has a suitable hydrophilic-hydrophobic ratio, resulting in a stronger enrichment in the tumor site.

[0284] A subcutaneous tumor model of hepatocarcinoma was obtained by subcutaneous inoculation of HepG2 cells. HC-9-PEG10 (Example 9) was used as a photosensitizing drug, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-9-PEG10 in the tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 h. As shown in Figure 15 (f) shows that the drug molecule HC-1-PEG50 has a strong enrichment in the tumor site after several cycles in vivo by passive targeting for about 4 h after intravenous injection, because this derivative molecule contains 50 polyethylene glycol units, making the water solubility of the whole drug molecule better, and the whole molecule has a suitable hydrophilic-hydrophobic ratio, resulting in a stronger enrichment in the tumor site.

[0285] 2) Drug enrichment in head and neck facial tumor mice (brain cancer, head and neck cancer, tongue cancer, oral cancer, nasal cancer, brain glioma)

[0286] The brain cancer G442 cells were used to inoculate subcutaneous tumor to get the brain cancer subcutaneous mouse tumor model. HB-11-PEG6 (Example 11) was used as photosensitizer and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HB-11-PEG6 in the brain cancer tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As shown in Fig. 2(a), HB-11-PEG6 has a certain enrichment ability in the tumor site through passive targeting after several cycles in vivo by intravenous injection, which belongs to moderate enrichment. The reason is that the derivative molecule only contains 6 polyethylene glycol units, so the water solubility of the whole drug molecule is not enough, resulting in moderate enrichment in the tumor site. Figure 16

[0287] The head and neck cancer SCC2 cells were used to inoculate subcutaneous tumor to get the head and neck cancer subcutaneous mouse tumor model. HC-18-PEG8 (Example 15) was used as photosensitizer and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-18-PEG8 in the head and neck cancer tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As shown in Fig. 2(b), HC-18-PEG8 has very strong enrichment in the tumor site through passive targeting after several cycles in vivo by intravenous injection. The reason is that the derivative contains 8 polyethylene glycol units, so the water solubility of the whole drug molecule is good, and the hydrophilic-hydrophobic ratio is appropriate, resulting in strong enrichment in the tumor site. Figure 16

[0288] The tongue cancer TSCCa cells were used to inoculate subcutaneous tumor to get the tongue cancer subcutaneous mouse tumor model. HC-45 (Example 23) was used as photosensitizer and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-45 in the tongue cancer tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As shown in Fig. 2(c), HC-45 has moderate intensity enrichment in the tumor site through passive targeting after several cycles in vivo by intravenous injection. The reason is that the derivative molecule does not contain polyethylene glycol units, only contains one carboxyl group, and its water solubility is not enough, resulting in moderate intensity enrichment in the tumor site. Figure 16

[0289] The oral cancer CAL27 cells were used to inoculate subcutaneous tumor to get the oral cancer subcutaneous mouse tumor model. HC-57 (Example 27) was used as photosensitizer and injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-57 in the oral cancer tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4 hours. As shown in Fig. 2(d), HC-57 has moderate intensity enrichment in the tumor site through passive targeting after several cycles in vivo by intravenous injection. The reason is that the derivative molecule does not contain polyethylene glycol units, only contains one carboxyl group, and its water solubility is not enough, resulting in moderate intensity enrichment in the tumor site. Figure 16 ​​​(d) shows, HC-57 has a strong enrichment ability at the tumor site by passive targeting about 4h after several cycles in vivo by intravenous injection, the reason is that this derivative contains water-soluble groups such as carboxyl and quaternary ammonium salt, so that the water-solubility of the whole drug molecule is good, and the hydrophilic-hydrophobic ratio of the whole molecule is appropriate, thus leading to its strong enrichment at the tumor site.

[0290] The KB cell of nasopharyngeal carcinoma was used to inoculate subcutaneous tumor to obtain a nasopharyngeal carcinoma subcutaneous mouse tumor model. HB-65 (Example 29) was used as a photosensitizer, and was injected into tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HB-65 in tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4h. As shown in Figure 16 (e) shows, HB-65 has a strong enrichment ability at the tumor site by passive targeting about 4h after several cycles in vivo by intravenous injection, the reason is that this derivative contains hydrophilic groups such as quaternary ammonium salt and amide group, so that the water-solubility of the whole drug molecule is good, and the hydrophilic-hydrophobic ratio of the whole molecule is appropriate, thus leading to its strong enrichment at the tumor site.

[0291] The C6 cell of brain glioma was used to inoculate subcutaneous tumor to obtain a brain glioma subcutaneous mouse tumor model. HC-81-PEG16 (Example 33) was used as a photosensitizer, and was injected into tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-81-PEG16 in tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4h. As shown in Figure 16 (f) shows, HC-81-PEG16 has a strong enrichment ability at the tumor site by passive targeting about 4h after several cycles in vivo by intravenous injection, the reason is that this derivative contains 16 polyethylene glycol units, so that the water-solubility of the whole drug molecule is good, and the hydrophilic-hydrophobic ratio of the whole molecule is appropriate, thus leading to its strong enrichment at the tumor site.

[0292] 3) Drug enrichment in skin and urinary system tumor mice (basal cell carcinoma, squamous skin carcinoma, melanoma, cutaneous T-cell lymphoma, prostate cancer, bladder cancer)

[0293] The BCC cell of basal cell carcinoma was used to inoculate subcutaneous tumor to obtain a basal cell carcinoma subcutaneous mouse tumor model. HB-5-PEG10 (Example 5) was used as a photosensitizer, and was injected into tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HB-5-PEG10 in tumor-bearing mice was observed, and the fluorescence signal at the tumor site was collected at 4h. As shown in Figure 17(a) shows that HB-5-PEG10 has a strong enrichment in tumor site by passive targeting after several cycles in vivo by intravenous injection, and the reason is that this derivative contains 10 PEG units, which makes the whole drug molecule have good water solubility and suitable hydrophilic-hydrophobic ratio, thus leading to a strong enrichment in tumor site.

[0294] The squamous skin cancer PECA cells were used to inoculate subcutaneous tumors to obtain a squamous skin cancer subcutaneous mouse model. HC-8-PEG12 (Example 8) was used as a photosensitizer, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HC-8-PEG12 in the tumor-bearing mice was observed, and the fluorescence signal at the tumor site at 4 hours was collected. As shown in Figure 17 (b) shows that HC-8-PEG12 has a strong enrichment in tumor site by passive targeting after several cycles in vivo by intravenous injection, and the reason is that this molecule contains 8 PEG units, which makes the whole drug molecule have good water solubility and suitable hydrophilic-hydrophobic ratio, thus leading to a strong enrichment in tumor site.

[0295] The melanoma B16 cells were used to inoculate subcutaneous tumors to obtain a melanoma subcutaneous mouse tumor model. HB-64 (Example 28) was used as a photosensitizer, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HB-64 in the tumor-bearing mice was observed, and the fluorescence signal at the tumor site at 4 hours was collected. As shown in Figure 17 (c) shows that HB-64 has a strong enrichment in tumor site by passive targeting after several cycles in vivo by intravenous injection, and the reason is that this molecule, although does not contain PEG units, contains water-soluble groups such as carboxylate and quaternary ammonium salt, which makes the whole drug molecule have good water solubility and suitable hydrophilic-hydrophobic ratio, thus leading to a strong enrichment in tumor site.

[0296] The cutaneous T-cell lymphoma HH cells were used to inoculate subcutaneous tumors to obtain a lymphoma subcutaneous mouse tumor model. HB-89-PEG16 (Example 36) was used as a photosensitizer, and was injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of HB-89-PEG16 in the tumor-bearing mice was observed, and the fluorescence signal at the tumor site at 4 hours was collected. As shown in Figure 17 (d) shows that HB-89-PEG16 has a strong enrichment in tumor site by passive targeting after several cycles in vivo by intravenous injection, and the reason is that this derivative contains 16 PEG units, which makes the whole drug molecule have good water solubility and suitable hydrophilic-hydrophobic ratio, thus leading to a strong enrichment in tumor site.

[0297] A subcutaneous prostate cancer mouse model was obtained by inoculating prostate cancer LNCaP cells into subcutaneous tumors. HC-90-PEG30 (Example 37) was used as a photosensitizing agent and injected into 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 fluorescence signals at the tumor site were collected after 4 hours. Figure 17 As shown in (e), after several cycles of intravenous injection, HC-90-PEG30 showed strong enrichment at the tumor site after about 4 hours of passive targeting. This is because the molecule contains 30 polyethylene glycol units, and the entire drug molecule has good water solubility and a suitable ratio of hydrophilicity to hydrophobicity, which leads to its strong enrichment at the tumor site.

[0298] A subcutaneous mouse model of bladder cancer was obtained by inoculating bladder cancer MBT-2 cells into subcutaneous tumors. HC-91-PEG16 (Example 38) was used as a photosensitizing agent and injected into 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 fluorescence signals at the tumor site were collected after 4 hours. The animal fluorescence imaging results are shown below. Figure 17 As shown in (f), after several cycles of intravenous injection, the drug molecule HC-91-PEG16 exhibits a strong accumulation capacity at the tumor site through passive targeting for approximately 4 hours. This is because the derivative molecule contains 16 polyethylene glycol units, resulting in good water solubility and a suitable hydrophilic-hydrophobic ratio, which leads to its strong accumulation at the tumor site.

[0299] The above Figures 15-17 The results show that the bamboo red fungicide derivatives provided in this invention can be administered to tumor-bearing mice via tail vein injection. In vivo imaging of small animals revealed that these drugs produced ultra-strong or moderate fluorescence at the tumor sites in mice, indicating that these derivatives can be used as photosensitizing drugs for fluorescence imaging at tumor sites in tumor-bearing mice. This can be used to guide the surgical removal of tumor boundaries using fluorescence-mediated materials. The tumors include: esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, glioma, basal cell carcinoma, squamous cell carcinoma, melanoma, cutaneous T-cell lymphoma, prostate cancer, and bladder cancer.

[0300] For the water-soluble slightly worse bamboo red fungus derivative in the application, intratumoral injection can be used for administration. The bamboo red fungus derivative is dissolved in DMSO (dimethyl sulfoxide), and physiological saline is used for dilution by 5-10 times for intratumoral injection. Taking the basal cell carcinoma (BCC cell) subcutaneous mouse tumor model as an example, the derivative (HB-28, Example 19) is used as a photosensitizer, and is injected into the tumor-bearing mouse at a dose of 10 mg / kg by intratumoral injection. The fluorescence imaging behavior of the tumor-bearing mouse in the tumor is observed, and the fluorescence signal of the tumor at 4 hours is collected by using a small animal imaging system. As shown in Fig. 18(a), the drug molecule HB-28 is injected into the tumor cells and tissues by intratumoral injection, and has a strong enrichment capacity at the tumor. Figure 18 (a), the drug molecule HB-28 is injected into the tumor cells and tissues by intratumoral injection, and has a strong enrichment capacity at the tumor.

[0301] The application also discloses the enrichment of other water-soluble slightly worse derivatives in different tumor cells by intratumoral administration. By using a similar administration method, the fluorescence imaging behavior of the tumor-bearing mouse in the tumor is observed, and the fluorescence signal of the tumor at 4 hours is collected by using a multispectral small animal in vivo imaging system. For example, the fluorescence imaging of the photosensitizer HC-36 (Example 20) in the squamous skin cancer PECA cell mouse in Fig. 18(b); the fluorescence imaging of the photosensitizer HB-45 (Example 23) in the melanoma B16 cell mouse in Fig. 18(c); the fluorescence imaging of the photosensitizer HC-73 (Example 31) in the skin T cell lymphoma HH cell mouse in Fig. 18(d); the fluorescence imaging of HC-77 (Example 32) in the lung cancer cell A549 cell mouse in Fig. 18(e); and the fluorescence imaging of HC-80 (Example 32) in the cholangiocarcinoma cell MCC cell mouse in Fig. 18(f). From the fluorescence imaging of the photosensitizer in the tumor-bearing mouse, it can be seen that the photosensitizer has a strong enrichment capacity at the tumor. Figure 18 It can be seen that the photosensitizer HB-28 has a strong enrichment capacity at the tumor in the basal cell carcinoma, the photosensitizer HC-36 has a strong enrichment capacity at the tumor in the squamous skin cancer, the photosensitizer HB-45 has a strong enrichment capacity at the tumor in the melanoma, the photosensitizer HC-73 has a strong enrichment capacity at the tumor in the skin T cell lymphoma, the photosensitizer HC-77 has a strong enrichment capacity at the tumor in the lung cancer, and the photosensitizer HC-80 has a strong enrichment capacity at the tumor in the cholangiocarcinoma.

[0302] By changing the molecular structure and different substituents, the various bamboo red fungus derivatives of the application also have similar enrichment effects on other different tumors. The derivatives have a good drug enrichment in the esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, brain glioma, basal cell carcinoma, squamous skin cancer, skin T cell lymphoma, melanoma, prostate cancer, and bladder cancer mice.

[0303] The derivatives of the present application not only have good enrichment in the subcutaneous tumor of mice, but also have good drug enrichment effect in the orthotopic tumor of mice. In order to study the in situ enrichment of such derivatives in tumors, the present experiment inoculates glioma C6 cells in the brain of mice to obtain an orthotopic model of glioma for fluorescence imaging. The derivative HB-3-PEG12 in Example 3 is used as a photosensitizing drug, and is injected into the tumor-bearing mice through the tail vein at a dose of 10 mg / kg. The fluorescence imaging behavior of the tumor-bearing mice is observed, and the fluorescence signals of the tumor site at 0, 2, 3.5 and 5 hours are collected by using a small animal living imaging system. As shown in Figure 23, the fluorescence signal of the tumor area gradually increases with time, and reaches the highest at 5h, indicating that the photosensitizing drug HB-3-PEG12 is highly enriched in the tumor site, and there is no drug enrichment in the non-tumor area of the brain. The photosensitizing drug has very good targeting enrichment effect on glioma. In addition, the brain of the blank mice without drug administration has no fluorescence. Figure 19

[0304] The present application aims at the lack of mediated drugs to locate and guide the resection of tumors in the surgery of glioma. It is found that such derivatives can specifically accumulate in glioma tissue, and there is no photosensitizer accumulation in the brain area without tumors, which has very good tumor targeting enrichment effect. At this time, the tumor tissue is irradiated with light of a specific wavelength to 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 produce singlet oxygen under photosensitive conditions, which leads to tumor cell damage, which is used for photodynamic therapy of glioma, and has no damage to normal tissues. The derivatives provided by the present application not only have good enrichment effect on glioma, but also have good drug enrichment on other tumors, such as esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, colon cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, basal cell carcinoma, squamous skin cancer, melanoma, prostate cancer, and bladder cancer. The present application discloses a derivative of bambusin for the first time as a fluorescence-mediated material to guide the resection of solid tumors.

[0305] The above Figures 15-19 ​The results show that the derivatives provided in the present application can be used for fluorescence imaging of tumor-bearing mice. It can be seen that these photosensitive drugs produce super or moderate fluorescence imaging at the tumors of mice, indicating that such derivatives can be used as photosensitive drugs for fluorescence imaging of tumor cells in tumor-bearing mice, and can be used for fluorescence-mediated material to guide the resection of tumor boundaries in 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, brain glioma, basal cell carcinoma, squamous skin cancer, cutaneous T-cell lymphoma, melanoma, prostate cancer, bladder cancer. Other derivatives in this application also have the effect of in situ enrichment of the above tumor cells, can be used for fluorescence imaging of the above tumor cells in vivo, and can be used for material-mediated tumor boundary resection in surgery. The fluorescence imaging results of the above part of the bambusin derivatives in the tumor are shown in Table 5.

[0306] Table 5: In vivo enrichment of part of the derivatives of the present application on digestive tract tumors, head and neck tumors, skin tumors, and genitourinary tumors

[0307]

[0308]

[0309] Example 46 Photodynamic therapy

[0310] In view of the fact that the bambusin derivatives of the present application can efficiently kill a variety of tumor cells in cell experiments, and can be well enriched in tumors in mice as photosensitive drugs, the inventors further tested the derivatives as photodynamic drugs to kill tumor cells in mice. Using mice as a model, lung cancer A549 cells were inoculated in mice to form tumors, and when the tumor volume reached 200 mm 3 , lung cancer animal models were obtained for photodynamic therapy. The photosensitive drug HB-1-PEG8 (Example 1) was injected into the tumor-bearing mice at a dose of 10 mg / kg, and the in vivo fluorescence imaging behavior was observed. The fluorescence signal at the tumor site was collected for 4 h using a small animal live imaging system. 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. The second day after photodynamic therapy of lung cancer in mice, scab appeared, and gradually shrank to the 6th day, and completely disappeared after the 12th day, indicating that the lung cancer in mice was completely inhibited by photodynamic therapy Figure 20 . While the lung cancer-bearing mice in the control group (only injected with normal saline without photosensitive drug) were irradiated with a 635 nm laser at 0.1 W / cm 2Irradiating the tumor site with a 635nm laser for 10 minutes showed rapid tumor growth after 6 days, and the tumor was already severely affected after 12 days, with no effect on inhibiting tumor growth. Therefore, HB-1-PEG8, as a photosensitizing drug, is used in photodynamic therapy for lung cancer, and it has a significant effect in killing tumor cells and inhibiting tumor regeneration and recurrence.

[0311] The experiment also used a subcutaneous mouse model of glioma (inoculated with C6 cells) for photodynamic therapy. A dose of 10 mg / kg of the photosensitizing drug HB-6-PEG6 (Example 6) was injected via the tail vein into tumor-bearing mice, and the in vivo fluorescence imaging behavior was observed. Four hours later, 0.1 W / cm² of the drug was administered. 2 A 635nm laser was used to irradiate the tumor site for 10 minutes, and the data was recorded. For example... Figure 21 (b) On day 2 after photodynamic therapy (PDT) of glioma in mice, scabs appeared, gradually shrank on day 6, and completely disappeared after day 14, indicating that PDT completely inhibited the growth of gliomas in mice. In contrast, in the control group of glioma mice (injected only with saline, without photosensitizer), under the same experimental conditions, the tumors were growing rapidly after 6 days, showing no tumor-inhibiting effect. Figure 21 (a). Therefore, HB-6-PEG6, as a photosensitizing drug, is used in photodynamic therapy for glioma and has a significant effect in killing tumor cells and inhibiting tumor regeneration and recurrence.

[0312] The experiment also used a subcutaneous mouse model of melanoma (inoculated with B16 cells) for photodynamic therapy. HC-9-PEG8 (Example 9) was injected intravenously into tumor-bearing mice, and in vivo fluorescence imaging was observed. Four hours later, 0.1 W / cm² was used... 2 A 635nm laser was used to irradiate the tumor site for 10 minutes, and the data was recorded. For example... Figure 21 (c) On day 2 after photodynamic therapy (PDT) of melanoma in mice, scabs appeared, gradually shrank on day 6, and completely disappeared after day 14, indicating that PDT completely suppressed the tumors in the mice. In contrast, the tumor-bearing mice in the control group, under the same conditions but only injected with saline without the photosensitizer, showed rapid tumor growth after 6 days, with no tumor-inhibiting effect. Therefore, HC-9-PEG8, as a photosensitizer, can kill tumor cells and inhibit tumor regeneration and recurrence in PDT of melanoma.

[0313] 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 intravenously into tumor-bearing mice, and in vivo fluorescence imaging was observed. Four hours later, 0.1 W / cm² was used... 2 A 635nm laser was used to irradiate the tumor site for 10 minutes, and the data was recorded. For example... Figure 21(d), scab appeared on the 2nd day after the mice were treated with the photodynamic therapy, and gradually reduced on the 6th day, and completely disappeared after the 14th day, indicating that the tumor of the mice was completely inhibited after the photodynamic therapy. For the tumor mice in the control group, under the same experimental conditions, but only physiological saline was injected, and no photosensitive drug was injected, it was found that the tumor was growing rapidly after 6 days, and had no effect on inhibiting the tumor. Therefore, HB-63 as a photosensitive drug for photodynamic therapy of bladder cancer has obvious tumor killing effect and can inhibit tumor regeneration and recurrence.

[0314] The tumor tissues of the mice after different treatments were subjected to H&E staining and pathological analysis. 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 the mice found that compared with the control group, no mice died within 30 days after the photodynamic therapy, indicating that the photodynamic therapy had good anti-tumor effect on tumor-bearing mice.

[0315] The experiment used mice as a model, and also used cholangiocarcinoma (MCC cells) to inoculate subcutaneous tumors in mice to obtain a cholangiocarcinoma model for photodynamic therapy. Photosensitive drug HB-29 (Example 19) was injected intratumorally in tumor-bearing mice, and the fluorescence imaging behavior in the tumor-bearing mice was observed. After 4h, the tumor site was irradiated with a 635nm laser of 0.1W / cm 2 Figure 22 a). Scab appeared on the 2nd day after the mice were treated with the photodynamic therapy, and gradually reduced on the 6th day, and completely disappeared after the 14th day, indicating that the tumor of the mice was completely inhibited after the photodynamic therapy. For the tumor mice in the control group, under the same experimental conditions, but only physiological saline was injected, and no photosensitive drug was injected, it was found that the tumor was growing rapidly after 6 days, and had no effect on inhibiting the tumor. Therefore, HB-29 as a photosensitive drug in photodynamic therapy of cholangiocarcinoma has obvious tumor killing effect and can inhibit tumor regeneration and recurrence.

[0316] The experiment also used a subcutaneous colon cancer mouse model (inoculated with HCT116 cells) for photodynamic therapy. Photosensitive drug HB-45 (Example 23) was injected intratumorally in tumor-bearing mice, and the fluorescence imaging in vivo was observed. After 4h, the tumor site was irradiated with a 635nm laser of 0.1W / cm 2 Figure 22 ​​(b), scab appeared on the 2nd day after the photodynamic therapy, and gradually reduced on the 6th day, and completely disappeared after the 14th day, indicating that the colon cancer of the mice was completely inhibited by the photodynamic therapy. However, the control mice, under the same experimental conditions, but only injected with normal saline without injection of photosensitive drugs, were found to have rapidly growing tumors after 6 days without any effect of inhibiting tumors. Therefore, HB-45 as a photosensitive drug for photodynamic therapy of colon cancer has obvious killing of tumor cells and can inhibit tumor regeneration and recurrence.

[0317] A subcutaneous mouse model of oral cancer (inoculated with CAL27 cells) was used for photodynamic therapy. HC-73 (Example 31) was injected intratumorally into 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 / cm2for 10 min, and the data were recorded. As shown in 2 Fig. 2(c), scab appeared on the 2nd day after the photodynamic therapy, and gradually reduced on the 6th day, and completely disappeared after the 14th day, indicating that the oral cancer of the mice was completely inhibited by the photodynamic therapy. However, the control mice, under the same experimental conditions, but only injected with normal saline without injection of photosensitive drugs, were found to have rapidly growing tumors after 6 days without any effect of inhibiting tumors. Therefore, HC-73 as a photosensitive drug for photodynamic therapy of oral cancer has obvious killing of tumor cells and can inhibit tumor regeneration and recurrence. Figure 22

[0318] A subcutaneous mouse model of basal cell carcinoma (inoculated with BCC cells) was used for photodynamic therapy. HC-80 (Example 32) was injected intratumorally into 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 / cm2for 10 min, and the data were recorded. As shown in 2 Fig. 3(d), scab appeared on the 2nd day after the photodynamic therapy, and gradually reduced on the 6th day, and completely disappeared after the 14th day, indicating that the tumor of the mice was completely inhibited by the photodynamic therapy. However, the control mice, under the same experimental conditions, but only injected with normal saline without injection of photosensitive drugs, were found to have rapidly growing tumors after 6 days without any effect of inhibiting tumors. Therefore, HC-80 as a photosensitive drug for photodynamic therapy of basal cell carcinoma has obvious killing of tumor cells and can inhibit tumor regeneration and recurrence. Figure 22

[0319] ​​The bamboo red bacterium derivative of the present application can efficiently kill various tumor cells (such as lung cancer, bile duct cancer, colon cancer, oral cancer cells, basal cell cancer cells, melanoma cells, brain glioma cells, bladder cancer cells) in tumor-bearing mice. By injecting a certain dose of photosensitive drug into the tumor-bearing mice and irradiating with a laser of a certain intensity and wavelength, a good photodynamic therapy effect is achieved. Since the photosensitive drug has a killing effect on tumor cells, it is believed that a good photodynamic therapy effect can also be achieved for other tumor mice, such as esophageal cancer, gastric cancer, liver cancer, head and neck cancer, brain cancer, tongue cancer, nasal cancer, squamous skin cancer, skin T cell lymphoma, prostate cancer, etc. The other compounds disclosed in the present application, whether they are derivatives of bamboo red bacterium or deacetyl bamboo red bacterium, have a good photodynamic inactivation cell ability under photodynamic conditions. Therefore, the 2-amino substituted or ethylenediamine substituted derivatives of bamboo red bacterium disclosed in the present application can well treat tumors in mice.

[0320] Comparative Example 1

[0321] The photodynamic effect of the bamboo red bacterium derivative in the present application on HeLa is shown in Table 1. Figure 23 Under red light irradiation, 200 nM concentration of HB-1-PEG6 can kill more than 80% of HeLa cells, and the half lethal concentration IC 50 is about 120 nM; similarly, 200 nM concentration of HB-73 can kill more than 80% of HeLa cells, and the half lethal concentration IC 50 is about 80 nM; 200 nM concentration of HC-80 can kill more than 80% of HeLa cells, and the half lethal concentration IC 50 is about 80 nM.

[0322] Under the same conditions, 50 nM concentration of HB-1-PEG6 can kill 80% of esophageal cancer cells AKR, gastric cancer cells MFC, lung cancer cells A549, liver cancer cells HCC, bile duct cancer cells MCC, and colon cancer cells HCT116, and the half lethal concentration IC 50 is about 20-30 nM (Table 2); under the same conditions, 50 nM concentration of 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 brain glioma cells C6, and the half lethal concentration IC 50 is about 20-30 nM (Table 3); under the same conditions, 50 nM concentration of HC-80 can kill 85% of basal cell cancer cells BCC, squamous skin cancer cells PECA, melanoma cells B16, prostate cancer cells LNCaP, and bladder cancer cells MBT-2, and the half lethal concentration IC50 The value is about 20-30 nM (Table 4). Therefore, from Figure 23 As can be seen from Tables 2-4, the phototoxic effect of the porphyra derivative of the present application on the above tumor cells is significantly higher than that on HeLa cells.

[0323] Obviously, the above embodiments of the present application are only examples for clearly illustrating the present application, and are not intended to limit the embodiments of the present application. Based on the above description, other different forms of changes or variations can be made by those skilled in the art, and it is impossible to enumerate all the embodiments here. Any changes or variations falling within the scope of the technical solutions of the present application are still within the protection scope of the present application.

[0324] It should be noted that the porphyra derivative to be protected in the present application contains two enol tautomers, and the chemical structures of the two isomers are shown in formula (I) and formula (I'), which are of course within the protection scope. For the sake of simplicity, only one enol tautomer is listed in all the embodiments of the present application, and the other enol tautomer and its corresponding general structure are described in detail in the description, and the structure is of course within the protection scope. In addition, the general structure of the porphyra derivative in the present application contains a polyethylene glycol unit (PEGn), and the number n of the unit is any integer between 1 and 50, and the corresponding chemical structure is of course within the protection scope. For the sake of simplicity, only some integers are listed in all the embodiments of the present application, and the corresponding general structure of the rest is described in detail in the description, and the structure is of course within the protection scope. Any range described in the present application includes the end value and any numerical value between the end values, and any sub-range formed by the end value or any numerical value between the end values.

[0325] The embodiments of the present application are described above. However, the present application is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. The use of a bamboo red fungicide derivative or a mixture thereof in the preparation of a photodynamic antitumor drug, wherein, The tumor is esophageal cancer, gastric cancer, lung cancer, liver cancer, cholangiocarcinoma, colon cancer, brain cancer, head and neck cancer, tongue cancer, nasal cancer, oral cancer, brain glioma, basal cell carcinoma, squamous skin cancer, melanoma, cutaneous T-cell lymphoma, prostate cancer, bladder cancer, and the bambuterol derivative is a compound as shown in formula (I), an isomer, an isotopically labeled compound, a pharmaceutically acceptable salt or a solvate thereof. The structural formula of R1 in formula (I) is shown in formula (IV), and R2 is -H or -COCH3. In formula (IV), 0≤m≤12, 0≤n≤500, 0≤p≤12, 0≤q≤12; m, n, p, q are zero or positive integers; Y is a linking group; Z is an end group; (OCH2CH2) n is a polyethylene glycol unit; The linking group Y in formula (IV) is O, NH, S, carboxylate, amide, sulfonate, sulfonamide, phenylene, or a cycloalkyl group with 3-12 carbon atoms. The cycloalkyl group with 3-12 carbon atoms includes substituted or unsubstituted cycloalkyl groups, and the substituent is an alkyl group with 1-12 carbon atoms. The end group Z in formula (IV) is hydrogen, an alkyl group with 1-12 carbon atoms, an alkoxy group with 1-12 carbon atoms, phenyl, hydroxyl, amino, sulfhydryl, carboxylic acid, sulfonic acid, pyridyl, quaternary ammonium salt or pyridine salt. When the end group Z is a quaternary ammonium salt, the three substituents on the quaternary ammonium salt are independently selected from an alkyl group with 1-12 carbon atoms; and the anion in the quaternary ammonium salt is an anion allowed in pharmaceutical preparations. When the end group Z is a pyridine salt, the substituents on the pyridine ring are in ortho, meta or para position; the group connected to the pyridine N in the pyridine salt is a C1-8 alkyl group; and the anion in the pyridine salt is an anion allowed in pharmaceutical preparations.

2. Use according to claim 1, characterized in that, The linking group Y in the above formula (IV) is: -O-; -NH-; -S-; -COO-; -O-CO-; -CONH-; -NH-CO-; -SO3-; -C6H4- (phenyl); -C3H4- (cyclopropyl); -C4H6- (cyclobutyl); -C5H8- (cyclopentyl); -C5H7(CH3)- (methylcyclopentyl); -C6H 10 (cyclohexyl); -C6H9(CH3)- (methylcyclohexyl); -C7H 12 (cycloheptyl).

3. Use according to claim 1 or 2, characterized in that, The end group Z in the above formula (IV) is: -H; -CH3; -C2H5; -C4H9; -C6H 13 ; -OCH3; -OC2H5; -OC4H9; -OC6H 13 ; -C6H5; -OH, -NH2; -SH; -COOH; -COOCH3; -SO3H; -C5H4N; -C5H4N + ; -N + (CH3)3; -N + (C2H5)3; -N + (C6H 13 )3; -N + (CH3)2(C2H5); -N + (CH3)2(C6H 13 ); -N + (CH3)2(C8H 17 ).

4. Use according to any one of claims 1 to 3, characterized in that, The derivatives of formula I and formula I' are enol tautomers.

5. The use according to any one of claims 1 to 4, characterized in that, said R1is an alcohol of different chain length, and carboxylate formed with carboxyl polyethylene glycol: -(CH2) m -OH; -(CH2) m -OCH3; -(CH2) m -O-CO-CH2CH2-(OCH2CH2) n -OCH3[m is an integer between 1 and 8, n is an integer between 0 and 100] or R1is a carboxylic acid of varying chain length, and carboxylate or amide thereof with polyethylene glycol: -(CH2) m -COOH; -(CH2) m -COOCH3; -(CH2) m -CO-(OCH2CH2) n -OH; -(CH2) m -CO-(OCH2CH2) n -OCH3; -(CH2) m -CO-NH-CH2CH2-(OCH2CH2) n -OCH3[m is an integer between 1 and 8, n is an integer between 0 and 100] or R1 is a sulfonic acid group of different chain length, and a sulfonate or sulfonamide formed from a sulfonic acid group and a polyethylene glycol: -(CH2) m -SO3H; -(CH2) m -SO2-(OCH2CH2) n -OH; -(CH2) m -SO2-(OCH2CH2) n -OCH3; -(CH2) m -SO2-NH-CH2CH2-(OCH2CH2) n -OH; -(CH2) m -SO2-NH-CH2CH2-(OCH2CH2) n -OCH3[m is an integer between 1 and 8, n is an integer between 0 and 100] or R1is a thio-polyethylene glycol: -CH2CH2-SH; -CH2CH2-S-CH2CH2OH; -CH2CH2-S-CH2CH2OCH3; -CH2CH2-S-CH2CH2-(OCH2CH2) n -OH; or R1 is alkyl, amino, hydroxyl, or a phenyl, pyridyl containing substituent: -H; -CH3; -C2H5; -C3H7; -C4H9; -C5H 11 ; -C6H 13 ; -C8H 17 ; -NH2; -NHCH3; -NHC2H5; -OH; -CH2C6H5; -C5H4N; -CH2C5H4N; -(CH2)2C5H4N; -NHC6H5; -NHC5H4N; or R1is a cycloalkyl-containing substituent: -C3H5(cyclopropyl), -C4H7(cyclobutyl), -C5H9(cyclopentyl), -C6H 11 (cyclohexyl), -C6H 10 (CH3)(methylcyclohexyl), -C6H 10 (OH)(hydroxycyclohexyl), -C7H 13 (cycloheptyl), -CH2C6H 10 COOH, -CH2C6H 10 COOCH3, -CH2C6H 10 OH, -C6H 10 COOH; or R1is a substituted cyclohexane (-C6H 10 -OH; -CH2C6H 10 COOH; -CH2C6H 10 COOCH3; -CH2C6H 10 OH; -C6H 10 COOH); or R1is a quaternary ammonium salt containing substituent: -(CH2) m -N + (CH3)3; -(CH2) m -N + (CH3)2(C2H5); -(CH2) m -N + (CH3)2(C3H7); -(CH2) m -N + (CH3)2(C4H9); -(CH2) m -N + (CH3)2(C5H 11 ); -(CH2)3-N + (CH3)2(C6H 13 ); -(CH2) m -N + (CH3)2(C8H 17 ); -(CH2) m -N + (CH3)2(C 10 H 21 ); -(CH2) m -N + (CH3)2(C 12 H 25 ); -(CH2) m -O-CO-(CH2)2-N + (CH3)3; -(CH2) m -O-CO-(CH2)3-N + (CH3)3; -(CH2) m -O-CO-(CH2)4-N + (CH3)3; -(CH2) m -O-CO-(CH2)5-N + (CH3)3; -(CH2) m -O-CO-(CH2)6-N + (CH3)3; -(CH2) m -COO-(CH2)2-N + (CH3)3; -(CH2) m -COO-(CH2)3-N + (CH3)3; -(CH2) m -COO-(CH2)4-N + (CH3)3; -(CH2) m -COO-(CH2)5-N + (CH3)3; -(CH2) m -CONH-(CH2)2-N + (CH3)3; -(CH2) m -CONH-(CH2)2-N + (CH3)3; -(CH2) m -CONH-(CH2)3-N + (CH3)3; -(CH2) m -CONH-(CH2)4-N + (CH3)3[m is an integer between 1 and 8].

6. Use according to claim 1, characterized in that, Preferably, the structure of the bambuterol derivative is as follows: n is an integer between 1 and 100.

7. The use according to any one of claims 1 to 6, characterized in that, The tumor is esophageal cancer cell, gastric cancer cell, lung cancer cell, liver cancer cell, cholangiocarcinoma cell, colon cancer cell, head and neck cancer cell, brain cancer cell, tongue cancer cell, nasal cancer cell, oral cancer cell, brain glioma cell, basal cell carcinoma cell, squamous skin cancer cell, cutaneous T-cell lymphoma, melanoma cell, prostate cancer cell, bladder cancer cell.

8. Use according to any one of claims 1 to 7, characterized in that, The drug is a photodynamic drug or a fluorescence-mediated drug.

9. Use according to any one of claims 1 to 8, characterized in that, The pharmaceutically acceptable salt includes a salt formed by esterification of the compound of formula (I) with 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, and then with an inorganic base, including sodium, potassium, calcium, aluminum and ammonium salts, or with an organic base, including methylamine, ethylamine and ethanolamine salts; or a salt formed by esterification of the compound of formula (I) with a basic amino acid selected from lysine, arginine and ornithine, and then with an inorganic acid selected from hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid and phosphoric acid, or with an organic acid selected from formic acid, acetic acid, picric acid, methanesulfonic acid and ethanesulfonic acid.

10. Use of a compound of formula (I), an isomer, an isotopically labeled compound, a pharmaceutically acceptable salt or a solvate thereof in any one of claims 1-9 in the preparation of a fluorescence-mediated drug for guiding the boundary resection of a tumor.

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