A 2-position polyethylene glycol substituted water-soluble derivative of spongistatin and its use in treating tumors

Abstract

The application discloses a water-soluble polyethylene glycol modified bambusin derivative and application thereof in photodynamic treatment of tumors. The polyethylene glycol modified bambusin derivative prepared in the application has a maximum absorption wavelength of about 580 nm, and a molar extinction coefficient of 15000-20000 M-1cm-1, and has a strong light absorption capacity in a phototherapy window. The polyethylene glycol modified bambusin derivative prepared in the application can efficiently generate singlet oxygen and other active oxygen species under photosensitive conditions, and can efficiently inactivate various tumor cells. The polyethylene glycol modified bambusin derivative prepared in the application has good water solubility, and has a solubility of more than 10 mg / mL in physiological saline. The derivative can be used as a photosensitive drug to enter the body through tail vein administration, and can be used for photodynamic drug treatment of various tumors. Under the same conditions, the polyethylene glycol substituted bambusin derivative involved in the application has a higher photodynamic treatment effect than a commercial photosensitive drug of a porphyrin type.

Description

[0001] This application is a divisional application filed by the applicant with the China National Intellectual Property Administration on November 18, 2021, with patent application number 202111371173.0 and entitled "A water-soluble derivative of bamboo red fungin with polyethylene glycol substitution at the 2-position and its application in the treatment of tumors". Technical Field

[0002] This invention relates to the field of photosensitizer pharmaceutical technology, and more specifically, to a water-soluble polyethylene glycol-modified bamboo red fungus derivative, its preparation method, and its application in the treatment of tumors. Background Technology

[0003] Photodynamic therapy (PDT) is a novel method for treating tumors that emerged in the late 1970s. It has been officially approved in many countries, including the United States, Europe, Japan, and China, becoming a new technology for cancer treatment. PDT involves the ingestion of a photosensitizer within the body. Over a certain period, the photosensitizer accumulates in tumor cells and tissues. Irradiation of the tumor lesion with a light source of appropriate wavelength stimulates the photosensitizer to produce reactive oxygen species (ROS). These ROS (such as singlet oxygen, hydroxyl radicals, and superoxide radicals) damage tumor cells and destroy tumor tissue through photochemical reactions, achieving the goal of tumor treatment. Compared to traditional surgery, chemotherapy, and radiotherapy, PDT is more targeted in cancer treatment, selectively eliminating primary and recurrent tumors while reducing damage to normal cells. PDT is currently used to treat various tumors and has shown some efficacy in controlling locally growing tumors. Currently, photodynamic therapy (PDT) has been reported to treat various tumors, including skin tumors and precancerous lesions (such as basal cell carcinoma, squamous cell carcinoma, melanoma, etc.), head and neck tumors (such as nasopharyngeal carcinoma, laryngeal carcinoma, tongue cancer, oral cancer, etc.), brain tumors (glioma), reproductive tumors (prostate cancer, bladder cancer, cervical cancer, etc.), and digestive system tumors (cholangiocarcinoma, gastric cancer, lung cancer, liver cancer, colorectal cancer, pancreatic cancer). PDT not only has good efficacy for many primary tumors but also has unique technical advantages for many metastatic tumors. Studies have shown that PDT kills tumor cells directly through the generation of reactive oxygen species or by blocking the blood vessels that sustain tumor cell growth. Directly killing tumor cells utilizes the dual selectivity of PDT: photosensitizers selectively accumulate at tumor cells, and reactive oxygen species are generated near the lesion under irradiation with specific wavelengths of light, leading to the lesion's transformation and death. Anti-angiogenic photodynamic therapy (APPT) addresses the fact that tumor cells depend on blood supply for survival. APT can damage the tumor-associated vascular system, leading to ischemic death of the tumor. If light is applied during the peak concentration period of photosensitizers in blood vessels, it can cause microvascular damage, resulting in insufficient blood supply to the lesion and causing cell necrosis or apoptosis.

[0004] Photosensitizers, light sources, and oxygen are the three essential elements of photodynamic therapy (PDT). The key steps of PDT are: the specific aggregation of photosensitizers at the target site; the chemical reaction between light and photosensitizers to produce singlet oxygen and other active substances; and the targeted damage of these active substances to target cells. PDT has advantages over other treatment methods for tumors, as it can destroy tumor tissue while remaining largely non-toxic to normal tissues within permissible light doses, and it also has a therapeutic effect on the invasive margins of tumors. Photosensitizers used in PDT for tumors must meet the following conditions: 1) The photosensitizer has a single chemical composition and no significant toxic effects on the human body; 2) It can target tumor cells and tissues and remain within the tumor tissue, rarely entering normal cells; 3) The photosensitizer has deep penetration into tumor tissue to excitation light; 4) Under light irradiation, it can selectively damage tumor cells while causing minimal damage to normal cells and tissues; 5) It can be rapidly excreted from the body after treatment without other toxic side effects. Currently, photosensitizers used in tumor research and treatment mainly include hematoporphyrin derivatives (HpD), 5-aminolevulinic acid (5-ALA), talaporfin, temoporfin, and verteporfin. However, these photosensitizers all belong to the porphyrin class of compounds, and 5-aminolevulinic acid is also converted to protoporphyrin through in vivo biosynthesis. The main problem with porphyrin photosensitizers is the difficulty in separating their geometric isomers, making it difficult to obtain single-component photosensitizers. The relatively complex composition of these mixed photosensitizers is not conducive to subsequent drug metabolism and toxicological analysis. In addition, porphyrin compounds have a long retention time in vivo, requiring a long light-shielding period, which is not conducive to clinical use. In order to better utilize photodynamic therapy in the treatment of tumors, it is necessary to develop novel, highly efficient, low-toxicity, and stable photosensitizers with strong light absorption capacity within the phototherapy window.

[0005] Hypocrellin, a perylene quinone compound, is a natural plant-based photosensitizer produced by *Hypocrellus basilica*, a parasitic fungus found on arrow bamboo at altitudes above 3000 meters in Yunnan Province, my country. Natural hypocrellin is primarily composed of hypocrellin A (HA) (95%), with a small amount existing as hypocrellin B (HB) (5%). Hypocrellin exhibits strong absorption in the visible light region and a large molar extinction coefficient, and can efficiently generate reactive oxygen species under photosensitizing conditions. Previous studies have demonstrated that hypocrellin possesses advantages such as good phototoxicity, low dark toxicity, a well-defined structure, rapid in vivo metabolism, and no in vivo toxicity. However, natural hypocrellin is a hydrophobic compound with a primary absorption wavelength of 460 nm, requiring further modification to reach the phototherapy window. Previous studies have shown that the structure of bamboo red cinnamicin is easy to modify. The modified derivatives can meet the requirements of strong absorption of light in the phototherapy window (600-900nm) and water solubility that meets the needs of clinical intravenous injection. Therefore, bamboo red cinnamicin has broad application prospects as a photodynamic drug.

[0006] Research on the photodynamic therapy of bamboo red sclerosing agent for tumors is still in the research stage and there are few reports. Therefore, there is an urgent need to develop new bamboo red sclerosing agent photodynamic drugs for the treatment of tumor diseases. Summary of the Invention

[0007] To address the aforementioned technical problems, this invention provides a bamboo red fungicide derivative of formula (I), and its isomers, isotope labels, or pharmaceutically acceptable salts:

[0008]

[0009] In equation (I), R1 is -H or -COCH3;

[0010] The general structural formula of R2 is —NH—(CH2). p —X—Y—(CH2CH2O) n —Z;

[0011] Wherein, X is a linking group, selected from -C 3-8 Cycloalkyl-, -(CH2CH2O) m -;

[0012] Y is a linking group, selected from the following groups: -COO-, -O-CO-, -CONH-, -NHCO-, -COO-CH2-, -O-CO-CH2-, -O-CO-CH2O-, -CO-, -CO-CH2-, -CO-CH2O-. The left side of the above groups is connected to X, and the right side is connected to the polyethylene glycol unit.

[0013] Z is a terminal base, selected from -H and -C. 1-6 Alkyl, -C 1-6 Alkyl-COOH, -C 1-6 Alkyl-COO-C 1-6 Alkyl, -C 1-6 Alkyl-SO3H, -C 1-6 Alkyl-SO3-C 1-6 alkyl;

[0014] p is selected from 0 or 1; m is selected from 2 or 3;

[0015] -(CH2CH2O) n - represents polyethylene glycol units, and n is selected from integers between 1 and 100, for example, n is selected from integers between 1 and 50;

[0016] In one embodiment, X is selected from cyclohexyl, cyclopentyl, cyclobutyl, cyclopropyl, or -(CH2CH2O). m -, where m is selected from 2 or 3;

[0017] In one implementation, Y is selected from -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O-, -C(=O)NH-, -NHC(=O)-, -C(=O)-, -C(=O)-CH2- or -C(=O)-CH2O-;

[0018] Z is selected from -H, -CH3, -C2H5, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H.

[0019] According to an embodiment of the present invention, when X is cyclohexyl, Y is selected from -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O- or -C(=O)NH-;

[0020] When X is cyclopentyl, Y is selected from -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O- or -C(=O)NH-;

[0021] When X is cyclobutyl, Y is selected from -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O- or -C(=O)NH-;

[0022] When X is cyclopropyl, Y is selected from -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O- or -C(=O)NH-;

[0023] When X is -(CH2CH2O) m When -, m is selected from 2 or 3, and Y is selected from -C(=O)-, -C(=O)-CH2- or -C(=O)-CH2O-.

[0024] In one implementation, R2 is selected from:

[0025] -NH-CH2C6H 10 -COO-(CH2CH2O) n -Z、-NH-CH2C6H 10 -COO-CH2-(CH2CH2O) n -Z、

[0026] -NH-CH2C6H 10 -CO-NH-(CH2CH2O) n -Z、-NH-CH2C6H 10 -O-CO-(CH2CH2O) n -Z、

[0027] -NH-C6H 10 -COO-(CH2CH2O) n -Z、-NH-C6H 10 -CO-NH-(CH2CH2O) n -Z、

[0028] -NH-C6H 10 -COO-CH2-(CH2CH2O) n -Z、-NH-C6H 10 -O-CO-(CH2CH2O) n -Z、

[0029] -NH-C6H 10 -O-CO-CH2-(CH2CH2O) n -Z、-NH-C6H 10 -O-CO-CH2O-(CH2CH2O) n -Z、

[0030] -NH-C5H8-COO-(CH2CH2O) n -Z、-NH-C5H8-CO-NH-(CH2CH2O) n -Z、

[0031] -NH-C5H8-COO-CH2-(CH2CH2O) n -Z、-NH-C5H8-O-CO-(CH2CH2O) n -Z、

[0032] -NH-C5H8-O-CO-CH2-(CH2CH2O) n -Z、-NH-C5H8-O-CO-CH2O-(CH2CH2O) n -Z、

[0033] -NH-C4H6-COO-(CH2CH2O) n -Z、-NH-C4H6-CO-NH-(CH2CH2O) n -Z、

[0034] -NH-C4H6-COO-CH2-(CH2CH2O) n -Z、-NH-C4H6-O-CO-(CH2CH2O) n -Z、

[0035] -NH-C4H6-O-CO-CH2-(CH2CH2O) n -Z、-NH-C4H6-O-CO-CH2O-(CH2CH2O) n -Z、

[0036] -NH-C3H4-COO-(CH2CH2O) n -Z、-NH-C3H4-CO-NH-(CH2CH2O) n -Z、

[0037] -NH-C3H4-COO-CH2-(CH2CH2O) n -Z、-NH-C3H4-O-CO-(CH2CH2O) n -Z、

[0038] -NH-C3H4-O-CO-CH2-(CH2CH2O) n -Z、-NH-C3H4-O-CO-CH2O-(CH2CH2O) n -Z、

[0039] -NH-(CH2CH2O) m -CO-(CH2CH2O) n -Z、-NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -Z、

[0040] -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -Z、

[0041] In the above groups, m is selected from 2 or 3; each n may be the same or different, and is independently selected from integers from 1 to 100; Z is selected from -H, -CH3, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H; -C6H 10 - represents cyclohexyl, -C5H8- represents cyclopentyl, -C4H6- represents cyclobutyl, and -C3H4- represents cyclopropyl.

[0042] In one embodiment, when X is a cyclohexyl group, the -(CH2) group attached to the cyclohexyl group... p - and group Y are located at the para, ortho, or meta position; more preferably, both groups are located at the para position;

[0043] In one embodiment, when X is cyclopentyl, the group -(CH2) attached to the cyclopentyl group... p - and the Y group is located in the ortho or meta position;

[0044] In one embodiment, the terminal group Z is selected from -CH3, -H, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H;

[0045] In one implementation, R2 is, for example:

[0046] Each of the above n may be the same or different, and each is independently selected from an integer between 1 and 100;

[0047] For example, R2 is selected from:

[0048] In one implementation, R2 is, for example:

[0049] Each of the above n may be the same or different, and each is independently selected from an integer between 1 and 100;

[0050] For example, R2 is selected from

[0051] In one implementation, R2 is, for example:

[0052] In one implementation, R2 is, for example:

[0053] In one implementation, R2 is, for example:

[0054] In one implementation, R2 is, for example:

[0055] In one implementation, R2 is, for example:

[0056] In one implementation, R2 is, for example:

[0057] In one implementation, R2 is, for example:

[0058] In one implementation, R2 is, for example:

[0059] In one implementation, R2 is, for example:

[0060] In one implementation, R2 is, for example:

[0061] Each of the above n may be the same or different, and each is independently selected from an integer between 1 and 100.

[0062] In one embodiment, R2 is, for example, -NH-(CH2CH2O). m -CO-(CH2CH2O) n -CH3、-NH-(CH2CH2O) m -CO-(CH2CH2O) n -H, -NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2COOH, -NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2CH2COOH, -NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2CH2COOCH3, -NH-(CH2CH2O) m-CO-(CH2CH2O) n -CH2CH2SO3H, -NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -CH3、-NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -H, -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -CH3 or -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -H;

[0063] Each m is selected from 2 or 3; each n is the same or different and is selected independently from integers from 1 to 100;

[0064] For example, R2 is selected from -NH-(CH2CH2O)2-CO-(CH2CH2O)2-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)4-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)5-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)6-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)7-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)8-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)9-CH3, and -NH-(CH2CH2O)2-CO-(CH2CH2O) 10 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 12 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 14 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 16 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 18 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 20 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 24 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 32 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)36 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 40 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 50 -CH3 or -NH-(CH2CH2O)2-CO-(CH2CH2O) 100 -CH3.

[0065] The general structural formula of the bamboo red fungicide derivative described in formula (I) also includes the enol tautomer shown in formula (I'). Therefore, although this application only lists the structure of formula (I), the structure of formula (I') is also included within the scope of protection of this application. Those skilled in the art should not exclude the structure of formula (I') from the scope of protection of this application simply because this application does not list all structures of formula (I').

[0066]

[0067] The present invention also provides a method for preparing the derivative of formula (I), comprising the following steps:

[0068]

[0069] Wherein, R1, X, Y, Z, p, and n have the definitions described above; R3 is H, -COOH, -OH, and -NH2; R4 is H, -COOH, -OH, -CH2OH, -CH2COOH, -OCH2COOH, and -NH2;

[0070] 1) Reacting compound (III) with NH2-(CH2) p The -X-R3 reaction yields compound (II);

[0071] 2) The compound of formula (II) is combined with compound R4-(CH2CH2O) n The -Z reaction yields compound (I), wherein R3 and R4 react to form a linking group Y.

[0072] According to the present invention, in the preparation method:

[0073] When R3 is -COOH, equation (II) is combined with R4-(CH2CH2O) where R4 is -OH, -NH2, or -CH2OH. n -Z reacts to give the compound of formula (I), wherein the connecting bond Y is -COO-, -CONH- or -COO-CH2-;

[0074] When R3 is -OH, equation (II) is combined with R4-(CH2CH2O) where R4 is -COOH, -CH2COOH, or -OCH2COOH. n-Z reacts to give the compound of formula (I), wherein the connecting bond Y is -O-CO-, -O-CO-CH2- or -O-CO-CH2O-;

[0075] When R3 is -NH, equation (II) is combined with R4-(CH2CH2O) where R4 is -COOH, -CH2COOH, or -OCH2COOH. n -Z reacts to give the compound of formula (I), wherein Y is -NHCO-, -NHCO-CH2- or -NHCO-CH2O-;

[0076] When X is -(CH2CH2O) m When R3 is -H, equation (II) is combined with R4-(CH2CH2O) where R4 is -COOH, -CH2COOH, or -OCH2COOH. n -Z reacts to yield compound (I), where X is -(CH2CH2O). m -, Y is -CO-, -CO-CH2- or -CO-CH2O-.

[0077] In one embodiment, the reactions described in steps 1) and 2) are carried out in the dark under the protection of an inert gas, preferably argon or nitrogen.

[0078] In one embodiment, the compound of formula (III) in step 1) is basiloxane or deacetylated basiloxane.

[0079] In one embodiment, the compound of formula (III) described in step 1) and the compound NH2-(CH2) p The molar ratio of -X-R3 is 1:5 to 1:100, specifically 1:5, 1:10, 1:20, 1:40, 1:60, 1:80 or 1:100, and more preferably 1:60.

[0080] In one embodiment, the reaction temperature in step 1) is 50–120°C, more preferably 80°C.

[0081] In one embodiment, the reaction time in step 1) is 4 to 24 hours, more preferably 8 hours.

[0082] In one embodiment, the reaction in step 1) is carried out in a solvent, which is an organic solvent or a mixture of an organic solvent and water. The water content in the organic solvent and water mixture is 10 wt% to 95 wt%; the organic solvent is one or more of acetonitrile, tetrahydrofuran, pyridine, N,N-dimethylformamide, dimethyl sulfoxide, methanol, or ethanol. More preferably, the organic solvent is N,N-dimethylformamide, tetrahydrofuran, or acetonitrile; even more preferably, the organic solvent and water mixture is a mixture of N,N-dimethylformamide and water, wherein the volume ratio of N,N-dimethylformamide to water is 1:1.

[0083] In one embodiment, the reaction in step 1) can also be carried out under alkaline conditions, wherein the alkaline conditions are pH = 10-14. The reagents used for the alkaline conditions are aqueous solutions of potassium hydroxide, sodium hydroxide, or potassium carbonate. More preferably, the reaction is carried out in a 1% aqueous solution of sodium hydroxide.

[0084] In one embodiment, the reaction liquid treatment process in step 1) is as follows: the reaction organic solvent is removed, the blue-black solid residue is dissolved in dichloromethane, washed three times with 5% dilute hydrochloric acid aqueous solution and washed once with water, the organic layer is dried with anhydrous magnesium sulfate, filtered, and the solvent is removed to obtain compound (II).

[0085] In one embodiment, the compound of formula (II) described in step 2) and compound R4-(CH2CH2O) n The molar ratio of -Z is 1:1 to 10, with 1:1 to 5 as an example, such as 1:2;

[0086] In one embodiment, the reaction in step 2) can be carried out under the action of a condensing agent, which can be dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt), or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI).

[0087] In one embodiment, the reaction in step 2) can be carried out in the presence of an organic base, which may be triethylamine, N,N-diisopropylethylamine or 4-dimethylaminopyridine (DMAP).

[0088] In one embodiment, the reaction in step 2) can be carried out in the presence of a solvent, which is an organic solvent, such as tetrahydrofuran, dioxane, dichloromethane, or chloroform.

[0089] In one embodiment, the reaction temperature in step 2) is 20–50°C, more preferably 25°C;

[0090] In one embodiment, the reaction time in step 2) is 4 to 24 hours, more preferably 24 hours.

[0091] In one embodiment, the separation and purification process of compound (I) in step 2) is as follows: the organic solvent of the reaction is removed, the blue-black solid residue is dissolved in dichloromethane, washed three times with 5% dilute hydrochloric acid aqueous solution and once with water, the organic layer is dried with anhydrous magnesium sulfate, filtered, and the solvent is removed to obtain the crude product. The obtained crude product is further separated by silica gel plate chromatography, with the developing solvent preferably being a mixture of ethyl acetate, diethylamine and ethanol in a volume ratio of 20:1:2, to obtain a 2-polyethylene glycol-substituted derivative of baicalein. The overall yield of the two-step reaction is 3-15%, and the product is a blue-black solid.

[0092] In this invention, the developing solvent used in silica gel column chromatography or silica gel plate chromatography is a conventional reagent, and 1% sodium tartrate, sodium citrate, or potassium dihydrogen phosphate is added to the stationary phase. Preferably, the developing solvent used in silica gel plate chromatography is a mixture containing ethyl acetate, diethylamine, and ethanol, wherein the volume ratio of ethyl acetate, diethylamine, and ethanol in the mixture is 20:1:1 to 20:1:5.

[0093] The present invention also provides a pharmaceutical composition comprising at least one of the compounds represented by formula (I) as described above, its isomers, isotopic labels, or pharmaceutically acceptable salts.

[0094] According to embodiments of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0095] According to embodiments of the present invention, the pharmaceutical composition is a photodynamic therapy drug and / or a fluorescence-mediated drug and / or a photothermal therapy drug, more preferably a photodynamic therapy drug for treating tumor diseases and / or a fluorescence-mediated drug and / or a photothermal therapy drug for guiding tumor resection.

[0096] The present invention also provides the use of the compound represented by formula (I), and at least one of its isomers, isotope labels or pharmaceutically acceptable salts in the preparation of a medicament.

[0097] According to an embodiment of the present invention, the drug is an antitumor drug, such as a photodynamic therapy drug and / or a fluorescence-mediated drug and / or a photothermal therapy drug.

[0098] According to embodiments of the present invention, the tumor is, for example, a digestive tract tumor, a head and neck facial tumor, a skin tumor, or a genitourinary system tumor; the digestive tract tumor includes esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, and colon cancer; the head and neck facial tumor includes head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, and glioma; the skin tumor includes basal cell carcinoma, squamous cell carcinoma, and melanoma; and the genitourinary system tumor includes prostate cancer and bladder cancer.

[0099] The present invention also provides a method for treating tumors, comprising administering to an individual in need a therapeutically effective amount of a compound of formula (I), at least one of its isomers, isotopic labels, or pharmaceutically acceptable salts.

[0100] The present invention also provides the use of compounds of formula (I), isomers thereof, isotopic labels or pharmaceutically acceptable salts as fluorescence-mediated drugs in guiding the border resection of tumors.

[0101] The present invention also provides a method for guiding tumor resection, comprising administering to an individual in need a therapeutically effective amount of a compound of formula (I), at least one of its isomers, isotopic markers, or pharmaceutically acceptable salts.

[0102] The maximum absorption wavelength of the bamboo red fungicide derivative described in this invention is around 580 nm, and the molar extinction coefficient can reach 15000-20000 nm. -1 cm -1 This invention discloses a water-soluble polyethylene glycol-modified bamboo red fungus derivative that exhibits strong light absorption capacity within the phototherapy window. Under photosensitized conditions, this derivative can efficiently generate singlet oxygen and other reactive oxygen species, effectively inactivating various tumor cells, such as esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, and colon cancer associated with the digestive tract; head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, and glioma associated with head, neck, and face cancer; basal cell carcinoma, squamous cell carcinoma, and melanoma associated with skin cancer; and tumors related to the reproductive and urinary systems, including prostate cancer and bladder cancer. Under the same conditions, the polyethylene glycol-substituted bamboo red fungus derivative photosensitizer described in this invention has a higher photodynamic inactivation ability for tumor cells than commercially available porphyrin-based photosensitizers (HpD and Ce6). Furthermore, this invention also discloses that such bamboo red fungus derivatives, in addition to generating reactive oxygen species under laser irradiation, can undergo significant photothermal conversion, with a photothermal conversion efficiency between 25-35%, making them suitable for photodynamic / photothermal synergistic tumor treatment. Under the same conditions, the combined effect of photodynamic / photothermal therapy is superior to that of photodynamic therapy alone. This application discloses for the first time that such bamboo red fungin derivatives possess excellent photothermal conversion properties.

[0103] The 2-position polyethylene glycol-substituted water-soluble derivative of the bamboo red fungin described in this invention has a solubility of more than 10 mg / mL in physiological saline. Such derivatives can be administered into the body via the tail vein as photosensitizing drugs, and can specifically accumulate in tumor cells or tissues. They can be used as photodynamic drugs to treat various tumors, or as fluorescence-mediated drugs to assist in the surgical resection of tumors.

[0104] Beneficial effects

[0105] 1) The bamboo red mycin raw material in this invention is extracted from natural products. The raw material is readily available, low in cost, can be prepared in large quantities, has few toxic side effects, and is easy to metabolize. The synthesis and separation method is simple, without expensive reaction raw materials and complex separation methods. The product is easy to separate, purify, and has a clear structure, overcoming the problems of porphyrin photosensitizers being difficult to separate and having complex composition and difficult to determine structure.

[0106] 2) The 2-position polyethylene glycol-substituted water-soluble derivative of bamboo red fungicide described in this invention has good water solubility and oil-water partition ratio, fully meeting the needs of clinical intravenous injection; it has strong red light absorption capacity during the phototherapy window and can efficiently generate reactive oxygen species under photosensitizing conditions; it has a higher photodynamic inactivation ability for tumor cells than porphyrin photosensitizers; furthermore, the compound described in this invention also has good photothermal conversion properties, and can generate heat to kill tumor cells under photothermal conditions. Therefore, the compound described in this invention can have a photodynamic / photothermal synergistic therapeutic effect on tumor cells, which is superior to the effect of photodynamic therapy alone.

[0107] 3) The 2-position polyethylene glycol-substituted water-soluble derivative of bamboo red fungicide described in this invention can be used as a photodynamic drug in the treatment of digestive tract tumors, head and neck facial tumors, skin tumors, and genitourinary system tumors. The digestive tract tumors include esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, and colon cancer; the head and neck facial tumors include head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, and glioma; the skin tumors include basal cell carcinoma, squamous cell carcinoma, and melanoma; and the genitourinary system tumors include prostate cancer and bladder cancer. This bamboo red fungicide derivative, as a photosensitizing drug, can efficiently kill various tumor cells via intravenous injection. A concentration of 50 nM of this photosensitizing drug can kill more than 90% of tumor cells while having virtually no effect on normal cells. This bamboo red fungicide derivative, as a fluorescence-mediated drug, can be used to guide tumor resection during surgery. Attached Figure Description

[0108] Figure 1 The general structural formula of the 2-position polyethylene glycol-substituted water-soluble derivative of the bamboo red mycotoxin of the present invention is shown.

[0109] Figure 2Synthetic routes for derivatives HB-1, HB-1-PEGn-CH3 (Example 2), and HB-1-NH-PEGn-CH3 (Example 12) are shown.

[0110] Figure 3 The synthetic route diagrams for derivatives HB-8 and HB-8-PEGn-CH3 (Example 28) are shown.

[0111] Figure 4 (a) Comparison of absorption spectra of commercial photosensitizers PpIX and Ce6. Figure 4 (b) is a comparison of the absorption spectra of HB, HB-1-PEG8-CH3 (Example 2), and HC-1-PEG8-CH3 (Example 7).

[0112] Figure 5 (a) is a diagram showing the interaction between HB-1-PEG16-CH3 (Example 2) and a singlet oxygen scavenger; Figure 5 (b) is a diagram showing the interaction between HB-1-PEG16-CH3 and the superoxide radical scavenger.

[0113] Figure 6 (a) Photodegradation curve of HB-1-PEG8-CH3 (Example 2); Figure 6 (b) is the photodegradation curve of HC-1-PEG8-CH3 (Example 7); Figure 6 (c) Photodegradation curve of the standard reference compound, Rose Red (RB); Figure 6 (d) is a comparison diagram of the photodegradation of HB-1-PEG8-CH3, HC-1-PEG8-CH3 and RB.

[0114] Figure 7 (a) The derivatives of this invention and commercial photosensitizers at 30 mW / cm 2 The photostability comparison graphs of the lasers after 30 min of illumination are shown. Curve (i) shows the photostability of HB-1-PEG16-CH3 (Example 2); curve (ii) shows the photostability of HC-1-PEG16-CH3 (Example 7); curve (iii) shows the photostability of the commercial photosensitizer Ce6; and curve (iv) shows the photostability of the commercial photosensitizer HpD.

[0115] Figure 7 (b) The derivatives of this invention and commercial photosensitizers at 30mW / cm 2 The photostability comparison graphs of laser illumination for 30 min are shown, where curves (i) and (ii) show the photostability of HB-8-PEG8-CH3 and HC-8-PEG8-CH3 (Example 28), respectively; curves (iii) and (iv) show the photostability of commercial photosensitizers Ce6 and HpD, respectively.

[0116] Figure 8 (a) is a comparison graph of the pH stability of the derivative of the present invention and commercial hematoporphyrin, wherein curve (i) shows the pH stability of the commercial photosensitizer HpD; curves (ii) and (iii) show the pH stability of HB-1-PEG24-CO2CH3 (Example 5) and HC-1-PEG24-CO2CH3 (Example 10), respectively.

[0117] Figure 8 (b) is a comparison graph of the pH stability of the derivative of the present invention and the commercial photosensitizer HpD; wherein curve (i) shows the pH stability of HpD; curves (ii) and (iii) show the pH stability of HB-8-PEG16-H (Example 29) and HC-8-PEG16-H (Example 29), respectively.

[0118] Figure 9 (a) HB-1-PEG16-CH3 (Example 2) under different laser intensities (0.1–0.8 W / cm²). -2 (a) Photothermal change curves under irradiation (temperature change of the solution within 10 minutes); (b) HB-1-PEG16-CH3, HC-1-PEG16-CH3 (Example 7), HB-1-PEG12-H (Example 3), and HC-1-PEG12-H (Example 8) at 0.8 W cm⁻¹ -2 Photothermal change curve under laser irradiation (temperature change of solution within 10 minutes).

[0119] Figure 10 Confocal fluorescence imaging of HB-1-PEG16-CH3 in C6 cells was prepared for Example 2 of this invention; wherein... Figure 10 (a) shows the overlay image of dark and bright fields. Figure 10 (b) shows the dark field image. Figure 10 (c) shows the bright field image.

[0120] Figure 11 The images show a comparison of the dark toxicity (a) and phototoxicity (b) of different concentrations of commercial HpD on esophageal cancer AKR cells, and the effects of HB-1-PEG8-CH3 on esophageal cancer AKR cells and HB-1-PEG16-CH3 on gastric cancer MFC cells in Example 2.

[0121] Figure 12 The images show a comparison of the dark toxicity (a) and phototoxicity (b) of different concentrations of commercial HpD on lung cancer cells A549, and the effects of HC-1-PEG8-CH3 on lung cancer cells A549 and HC-1-PEG16-CH3 on liver cancer cells HCC, as well as examples 7.

[0122] Figure 13 The image shows a comparison of the dark toxicity (a) and phototoxicity (b) of different concentrations of commercial HpD on cholangiocarcinoma cells MCC, and the effects of HB-8-PEG8-CH3 on cholangiocarcinoma cells MCC and HB-8-PEG16-CH3 on colon cancer cells HCT116 in Example 28.

[0123] Figure 14 Phototoxicity graphs of commercially available HpD at different concentrations and the compounds of this invention are shown:

[0124] (a) Comparison of phototoxicity of commercial HpD against brain cancer cells G442, comparison of phototoxicity of HC-8-PEG2-CH3 in Example 28 against brain cancer cells G442, and comparison of phototoxicity of HC-8-PEG8-CH3 against head and neck cancer cells SCC2.

[0125] (b) Phototoxicity diagrams of commercial HpD against tongue cancer cells TSCCa, HC-8-PEG16-H from Example 29 against tongue cancer cells TSCCa, and phototoxicity diagrams of HC-8-PEG32-H against nasal cancer cells KB.

[0126] (c) Phototoxicity graphs of commercial HpD against oral cancer cells CAL27, HC-8-PEG2-SO3H from Example 31 against oral cancer cells CAL27, and phototoxicity comparison graph of HC-8-PEG8-SO3H against glioma cells C6.

[0127] Figure 15 The following graphs show the photodynamic effects of different concentrations of bamboo red fungus derivatives on tumor cells:

[0128] (a) Comparison of phototoxicity of HB-1-PEG4-H in Example 3, HB-1-PEG16-CO2H in Example 4, and HB-1-PEG24-CO2CH3 in Example 5 to basal cell carcinoma (BCC) cells;

[0129] (b) is a comparison diagram of the phototoxicity of HC-1-PEG2-H in Example 8, HC-1-PEG24-SO3H in Example 11, and HB-1-NH-PEG4-CH3 in Example 12 to melanoma B16 cells.

[0130] (c) Comparison of phototoxicity of HB-1-NH-PEG16-H in Example 13, HC-1-NH-PEG4-CH3 in Example 17, and HC-1-NH-PEG24-COOCH3 in Example 20 to squamous cell carcinoma PECA cells;

[0131] (d) is a comparison of the phototoxicity of HB-2-p-PEG4 in Example 22, HB-5-PEG10 in Example 25, and HB-6-PEG24 in Example 26 to prostate cancer LNCaP cells;

[0132] (e) is a comparison of the phototoxicity of HB-7-PEG4, HC-7-PEG16 in Example 27 and HB-8-PEG32-CO2CH3 in Example 30 to bladder cancer MBT-2 cells;

[0133] (f) is a comparison of the phototoxicity of HB-9-PEG16-CH3 in Example 32, HB-10-PEG2-CH3 in Example 34, and HB-11-PEG16 in Example 36 to basal cell carcinoma (BCC) cells.

[0134] Figure 16 The images show the photodynamic effects of HB-1-PEG8-CH3 in Example 2, HC-1-PEG16-CH3 in Example 7, and HC-1-NH-PEG4-CH3 in Example 17 on HeLa cells.

[0135] Figure 17 In vivo fluorescence imaging of the derivatives of the present invention in different tumor model mice 4 hours after administration via tail vein injection: (a) HB-1-PEG16-CH3 of Example 2, esophageal cancer AKR cells; (b) HC-1-PEG16-CH3 of Example 7, lung cancer A549 cells.

[0136] Terminology Definitions and Explanations

[0137] Unless otherwise defined, all technical terms herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Numerical ranges described in this specification and claims, when defined as “integers,” should be understood to include both endpoints of the range and every integer within that range. For example, “integers from 1 to 6” should be understood to include every integer of 1, 2, 3, 4, 5, and 6, and so on.

[0138] Term "C" 1-6 "Alkyl" should be understood as representing a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 6 carbon atoms. For example, "C 1-6 "Alkyl" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, 2-ethylbutyl, etc., or isomers thereof.

[0139] Term "C" 1-6 "Alkoxy" should be understood as -OC 1-6 Alkyl, wherein C 1-6 Alkyl groups have the above definition.

[0140] Term "C" 3-8 "Cycloalkyl" should be understood as referring to saturated monovalent cycloalkyl groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0141] It should also be noted that the bamboo red fungicide derivatives protected in this invention all contain two enol tautomers, and the chemical structures of the two isomers, such as formulas (I) and (I'), are naturally within the scope of protection. For simplicity, only one enol tautomer is listed in all embodiments of this invention; the other enol tautomer and its corresponding general structural formula are described in detail in the specification, and its structure is naturally within the scope of protection. Furthermore, the general structural formula of the bamboo red fungicide derivatives involved in this invention includes polyethylene glycol units (PEGn), where the number of units n is any integer between 1 and 100, and the corresponding chemical structures are naturally within the scope of protection. For simplicity, only some integers are listed in all embodiments of this invention; the general structural formulas corresponding to the remaining parts are described in detail in the specification, and their structures are naturally within the scope of protection. Any range described in this invention includes end values ​​and any values ​​between end values, as well as any sub-ranges formed by end values ​​or any values ​​between end values. Detailed Implementation

[0142] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments and accompanying drawings, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive and should not be construed as limiting the scope of protection of the present invention. In this invention, unless otherwise specified, the experimental methods are conventional methods. Unless otherwise specified, the raw materials used are all obtainable from publicly available commercial sources; unless otherwise specified, all percentages are mass percentages; unless otherwise specified, M is mol / L. The bamboo red fungicide derivatives involved in this invention are shown in Examples 2-36, and their characterization results are shown in Table 1.

[0143] Example 1

[0144] The raw materials used in this invention are bamboo red fungus A (HA), bamboo red fungus B (HB), and deacetylated bamboo red fungus HC, the molecular structures of which are shown below:

[0145]

[0146] Example 2

[0147] Tranexamic acid-derived poly(ethylene glycol) compounds of different chain lengths (R1 = -COCH3, R2 = -NHCH2-C6H) 10 Preparation of -COO-PEGn-CH3)(PEGn is polyethylene glycol, n=2,8,12,16,20,24): Bambusa textilis HB (1 mmol), tranexamic acid H2N-CH2C6H 10 50 mmol of COOH and 200 mmol of NaOH were dissolved in 100 mL of a mixed solution of DMF and water (volume ratio 1:1). The mixture was heated to 120 °C under nitrogen protection and reacted in the dark for 6 h. After the reaction was complete, dilute hydrochloric acid was added to adjust the pH to weakly acidic, and a large amount of black solid precipitated out. The mixture was filtered to obtain the crude product HB-1. The crude product was then mixed with dicyclohexylcarbodiimide DCC (1 mmol) and 4-dimethylaminopyridine DMAP (1 mmol) in 50 mL of anhydrous dichloromethane and reacted separately with polyethylene glycol methyl esters (HO-PEGn-CH3, 1 mmol) of different chain lengths. The mixture was stirred at room temperature in the dark for 12 h. After the reaction was complete, the reaction solution was washed three times with 100 mL of dilute hydrochloric acid aqueous solution and distilled water, dried, and filtered. The crude product was separated by thin-layer chromatography with ethyl acetate and ethanol in a 5:1 ratio as the developing solvent to obtain the blue-black solid product HB-1-PEGn-CH3. The characterization results of the above amino-substituted products HB-1-PEGn-CH3 (n=2,8,12,16,20,24) are shown in Table 1, and their synthetic routes are as follows:

[0148]

[0149] Example 3

[0150] Tranexamic acid - HB derivatives with different chain lengths substituted with polyethylene glycol (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of -COO-PEGn-H): The preparation method is basically the same as in Example 2, except that polyethylene glycol (HO-PEGn-H) with different chain lengths is used to replace polyethylene glycol methyl ether to obtain the blue-black solid product HB-1-PEGn-H (n=4,12,18).

[0151] Example 4

[0152] Tranexamic acid - HB derivatives with different chain lengths substituted with polyethylene glycol (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of HO-PEGn-CH2COOH: The preparation method is the same as in Example 2, except that HO-PEGn-CH2COOH with different chain lengths is used to replace polyethylene glycol methyl ether to obtain the blue-black solid product HB-1-PEGn-CO2H (n=2,8,16).

[0153] Example 5

[0154] Tranexamic acid - HB derivatives with different chain lengths substituted with polyethylene glycol (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of HB-1-PEGn-CO2CH3: The preparation method is basically the same as in Example 2, except that HO-PEGn-CH2CH2COOCH3 is used to replace polyethylene glycol methyl ether to obtain the blue-black solid product HB-1-PEGn-CO2CH3 (n=8,16,24).

[0155] Example 6

[0156] Tranexamic acid - HB derivatives with different chain lengths substituted with polyethylene glycol (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of HO-PEGn-CH2CH2SO3H: The preparation method is basically the same as in Example 2, except that ethylene glycol methyl ether is replaced with HO-PEGn-CH2CH2SO3H of different chain lengths to obtain the blue-black product HB-1-PEGn-SO3H (n=2,8,16).

[0157] The characterization results of the amino-substituted products in Examples 3-6 above are shown in Table 1, and their molecular structures are shown below:

[0158]

[0159] Example 7

[0160] Tranexamic acid - HC derivatives of polyethylene glycol with different chain lengths (R1 = -H, R2 = -NHCH2-C6H) 10 Preparation of HO-1-PEGn-CH3: The preparation method is basically the same as in Example 2, except that HC is used to replace HB and HO-PEGn-CH3 is used to replace polyethylene glycol methyl ether, yielding the blue-black solid product HC-1-PEGn-CH3 (n = 2, 8, 12, 16, 20, 24). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0161]

[0162] Example 8

[0163] Tranexamic acid - HC derivatives of polyethylene glycol with different chain lengths (R1 = -H, R2 = -NHCH2-C6H) 10Preparation of HC-1-PEGn-H: Using HC-1 synthesized in Example 7 as the raw material, it was reacted with polyethylene glycol (HO-PEGn-H) of different chain lengths. The reaction was carried out at room temperature and in the dark with stirring for 12 h. After the reaction was completed, 100 mL of dichloromethane was added to the reaction solution, and the mixture was washed once with 100 mL of dilute hydrochloric acid aqueous solution and three times with distilled water. The organic layer was dried with anhydrous magnesium sulfate and filtered. The crude product was separated by thin-layer chromatography with a 5:1 mixture of ethyl acetate and ethanol as the developing solvent to obtain the blue-black solid product HC-1-PEGn-H (n = 2, 12, 16).

[0164] Example 9

[0165] Tranexamic acid - HC derivatives of polyethylene glycol with different chain lengths (R1 = -H, R2 = -NHCH2-C6H) 10 Preparation of HO-PEGn-CH2COOH: Using HC-1 synthesized in Example 7 as raw material, the polyethylene glycol used is HO-PEGn-CH2COOH. The synthesis method is similar to the synthesis method of HC-1-PEGn-CH3 in Example 7, and a blue-black solid product HC-1-PEGn-CO2H (n=2,8,24) is obtained.

[0166] Example 10

[0167] Tranexamic acid - HC derivatives of polyethylene glycol with different chain lengths (R1 = -H, R2 = -NHCH2-C6H) 10 Preparation of HO-PEGn-CH2CH2COOCH3: Using HC-1 synthesized in Example 7 as the raw material, the polyethylene glycol used was HO-PEGn-CH2CH2COOCH3. The synthesis method was similar to the synthesis method of HC-1-PEGn-CH3 in Example 7, and a blue-black solid product HC-1-PEGn-CO2CH3 (n=2,16,24) was obtained.

[0168] Example 11

[0169] Tranexamic acid - HC derivatives of polyethylene glycol with different chain lengths (R1 = -H, R2 = -NHCH2-C6H) 10 Preparation of HO-PEGn-CH2CH2SO3H: Using HC-1 synthesized in Example 7 as raw material, the polyethylene glycol used is HO-PEGn-CH2CH2SO3H. The synthesis method is similar to the synthesis method of HC-1-PEGn-CH3 in Example 7, and a blue-black solid product HC-1-PEGn-SO3H (n=2,8,24) is obtained.

[0170] The characterization results of the amino-substituted products in Examples 8-11 above are shown in Table 1, and their molecular structures are shown below:

[0171]

[0172] Example 12

[0173] Tranexamic acid - HB derivatives with different chain lengths of amino-polyethylene glycol substitution (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of H₂N-PEGn-CH₃: The amino-substituted polyethylene glycol used was H₂N-PEGn-CH₃. The synthesis method was similar to that of HB-1-PEGn-CH₃ in Example 2, yielding a blue-black solid product HB-1-NH-PEGn-CH₃ (n = 4, 8, 16, 24). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0174]

[0175] Example 13

[0176] Tranexamic acid - HB derivatives with different chain lengths of amino-polyethylene glycol substitution (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of H2N-PEGn-H: Using HB-1 synthesized in Example 2 as the raw material, it was reacted with polyethylene glycol (H2N-PEGn-H, 0.5 mmol) of different chain lengths. The reaction was carried out at room temperature and in the dark with stirring for 12 h. After the reaction was completed, 100 mL of dichloromethane was added to the reaction solution, and the mixture was washed once with 100 mL of dilute hydrochloric acid aqueous solution and three times with distilled water. The organic layer was dried with anhydrous magnesium sulfate and filtered. The crude product was separated by thin-layer chromatography with a 5:1 mixture of ethyl acetate and ethanol as the developing solvent to obtain the blue-black solid product HB-1-NH-PEGn-H (n = 2, 16, 24).

[0177] Example 14

[0178] Tranexamic acid - HB derivatives with different chain lengths of amino-polyethylene glycol substitution (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of H2N-PEGn-CH2COOH: Using HB-1 synthesized in Example 2 as raw material, the polyethylene glycol used is H2N-PEGn-CH2COOH. The synthesis method is similar to the synthesis method of HB-1-PEGn-H in Example 3, and a blue-black solid product HB-1-NH-PEGn-CO2H (n=8,16,24) is obtained.

[0179] Example 15

[0180] Tranexamic acid - HB derivatives with different chain lengths of amino-polyethylene glycol substitution (R1=-COCH3, R2=-NHCH2-C6H)10 Preparation of H2N-PEGn-CH2CH2COOCH3: Using HB-1 synthesized in Example 2 as the raw material, the polyethylene glycol used was H2N-PEGn-CH2CH2COOCH3. The synthesis method was similar to the synthesis method of HB-1-PEGn-H in Example 3, and a blue-black solid product HB-1-NH-PEGn-CO2CH3 (n=2,8,16) was obtained.

[0181] Example 16

[0182] Tranexamic acid - HB derivatives with different chain lengths of amino-polyethylene glycol substitution (R1=-COCH3, R2=-NHCH2-C6H) 10 Preparation of H2N-PEGn-CH2CH2SO3H: Using HB-1 synthesized in Example 2 as the raw material, the polyethylene glycol used is H2N-PEGn-CH2CH2SO3H. The synthesis method is similar to the synthesis method of HB-1-PEGn-H in Example 3, and a blue-black solid product HB-1-NH-PEGn-SO3H (n=2,8,16) is obtained.

[0183] The characterization results of the amino-substituted products in Examples 13-16 above are shown in Table 1, and their molecular structures are shown below:

[0184]

[0185] Example 17

[0186] Tranexamic acid - HC derivatives of different chain lengths of amino-polyethylene glycol substitution (R1=-H, R2=-NHCH2-C6H) 10 Preparation of HC-1-NH-PEGn-CH3: The bamboo red fungicide used was HC, and the polyethylene glycol used was NH2-PEGn-CH3. The synthesis method was similar to that of HB-1-PEGn-CH3 in Example 2, yielding a blue-black solid product HC-1-NH-PEGn-CH3 (n = 4, 8, 16, 24). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0187]

[0188] Example 18

[0189] Tranexamic acid - HC derivatives of different chain lengths of amino-polyethylene glycol substitution (R1=-H, R2=-NHCH2-C6H) 10Preparation of H2N-PEGn-H: Using HC-1 synthesized in Example 7 as the raw material, it was reacted with polyethylene glycol (H2N-PEGn-H, 0.5 mmol) of different chain lengths. The reaction was carried out at room temperature and in the dark with stirring for 12 h. After the reaction was completed, 100 mL of dichloromethane was added to the reaction solution, and the mixture was washed once with 100 mL of dilute hydrochloric acid aqueous solution and three times with distilled water. The organic layer was dried and filtered. The crude product was separated by thin-layer chromatography with ethyl acetate and ethanol in a ratio of 5:1 as the developing solvent to obtain the blue-black solid product HC-1-NH-PEGn-H (n = 2, 8, 24).

[0190] Example 19

[0191] Tranexamic acid - HC derivatives of different chain lengths of amino-polyethylene glycol substitution (R1=-H, R2=-NHCH2-C6H) 10 Preparation of H2N-PEGn-CH2COOH: Using HC-1 synthesized in Example 7 as raw material, the polyethylene glycol used is H2N-PEGn-CH2COOH, similar to the synthesis method of HC-1-PEGn-CH3, to obtain the blue-black solid product HC-1-NH-PEGn-COOH (n=8,16,24).

[0192] Example 20

[0193] Tranexamic acid - HC derivatives of different chain lengths of amino-polyethylene glycol substitution (R1=-H, R2=-NHCH2-C6H) 10 Preparation of H2N-PEGn-CH2CH2COOCH3: Using HC-1 synthesized in Example 7 as the raw material, the polyethylene glycol used was H2N-PEGn-CH2CH2COOCH3. Similar to the synthesis method of HC-1-PEGn-CH3, the blue-black solid product HC-1-NH-PEGn-CO2CH3 (n=8,16,24) was obtained.

[0194] Example 21

[0195] Tranexamic acid - HC derivatives of different chain lengths of amino-polyethylene glycol substitution (R1=-H, R2=-NHCH2-C6H) 10 Preparation of H2N-PEGn-CH2CH2SO3H: Using HC-1 synthesized in Example 7 as raw material, the polyethylene glycol used is H2N-PEGn-CH2CH2SO3H, similar to the synthesis method of HC-1-PEGn-CH3, to obtain the blue-black solid product HC-1-NH-PEGn-SO3H (n=2,8,16).

[0196] The characterization results of the amino-substituted products in Examples 18-21 above are shown in Table 1, and their molecular structures are shown below:

[0197]

[0198] Example 22

[0199] HB derivatives of anthranilic acid with different chain lengths and polyethylene glycol substitutions (R1 = -COCH3, R2 = -NH-C6H) 10 Preparation of HO-PEGn-CH3: The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to that of HB-1-PEGn in Example 2, yielding the blue-black solid product HB-2-o-PEGn (n = 4, 8, 16). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0200]

[0201] m-aminocyclic acid-PEG derivatives of HB with different chain lengths (R1=-COCH3, R2=-NH-C6H) 10 Preparation of HO-PEGn-CH3: The polyethylene glycol used was HO-PEGn-CH3. The synthesis method was similar to that of HB-1-PEGn in Example 2, yielding a blue-black solid product HB-2-m-PEGn (n = 4, 8, 16). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0202]

[0203] p-aminocyclic acid-substituted polyethylene glycol derivatives of different chain lengths (R1=-COCH3, R2=-NH-C6H) 10 Preparation of HO-PEGn-CH3: The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to that of HB-1-PEGn in Example 2, yielding the blue-black solid product HB-2-p-PEGn (n = 4, 8, 16). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0204]

[0205] Example 23

[0206] Preparation of aminocyclobutyric acid-substituted HB or HC derivatives of different chain lengths with polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-C4H6-COO-PEGn-CH3): The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to that of HB-1-PEGn in Example 2, yielding bamboo red sclerotin derivatives HB-3-PEGn (n = 4, 8, 16) and deacetylated bamboo red sclerotin derivatives HC-3-PEGn (n = 4, 8, 16). The characterization results of the above-mentioned amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0207]

[0208] Example 24

[0209] Preparation of HB derivatives with different chain lengths of aminocyclopropanol and polyethylene glycol substitution (R1 = -COCH3, R2 = -NHCH2-C3H4-O-CO-PEGn-CH3): The polyethylene glycol used was HOOC-PEGn-CH3, and the synthesis method was similar to that of HB-1-PEGn in Example 2, yielding the blue-black solid product HB-4-PEGn (n = 4, 8, 16). The characterization results of the above amino-substituted products are shown in Table 1, and their synthetic routes are as follows:

[0210]

[0211] Example 25

[0212] Preparation of HB or HC derivatives of m-aminocyclopentanoic acid substituted with polyethylene glycol of different chain lengths (R1 = -COCH3, R2 = -NH-C5H8-COO-PEGn-CH3): The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to that of HB-1-PEGn in Example 2, yielding blue-black solid products HB-5-PEGn (n = 2, 10, 20) and HC-5-PEGn (n = 2, 10, 20). The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0213]

[0214] Example 26

[0215] p-Aminocyclohexanol-substituted HB or HC derivatives of polyethylene glycol with different chain lengths (R1 = -COCH3 or -H, R2 = -NH-C6H) 10Preparation of HOOC-PEGn-CH3): The polyethylene glycol used was HOOC-PEGn-CH3. The synthesis method was similar to that of HB-1-PEGn in Example 2, yielding HB-6-PEGn (n = 1, 8, 24) and deacetylated HC-6-PEGn (n = 1, 8, 24) derivatives of HB-1-PEGn. The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0216]

[0217] Example 27

[0218] p-Aminocyclohexanol-substituted HB or HC derivatives of polyethylene glycol with different chain lengths (R1 = -COCH3 or -H, R2 = -NH-C6H) 10 Preparation of HOOC-CO-CH2-PEGn-CH3: The raw materials used were HB-6 or HC-6 prepared in Example 26, and the polyethylene glycol used was HOOC-CH2O-PEGn-CH3, yielding bamboo red pigment derivatives HB-7-PEGn (n = 4, 8, 16) and deacetylated bamboo red pigment derivatives HC-7-PEGn (n = 4, 8, 16). The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0219]

[0220] Example 28

[0221] Preparation of ethanolamine-substituted HB derivatives with different chain lengths of polyethylene glycol (R1 = -COCH3, R2 = -NH-(CH2CH2-O)2-CO-PEGn-CH3) (n = 2, 8, 16, 32): HB (1 mmol), glycolamine NH2-CH2CH2-O-CH2CH2-OH (50 mmol), and NaOH (100 mmol) were dissolved in 100 mL of a mixed solution of DMF and water (volume ratio 1:1). After thorough mixing, the mixture was heated to 120 °C under nitrogen protection and stirred in the dark for 6 h. After the reaction was complete, dilute hydrochloric acid was added to adjust the pH of the reaction solution to weakly acidic, and a large amount of black solid precipitated. The precipitate was collected by filtration to obtain the crude product HB-8. The crude product was added to DCC (1 mmol) and DMAP (1 mmol) and dissolved in 50 mL of anhydrous dichloromethane. The solutions were then reacted with polyethylene glycol methyl esters (HO-PEGn-OCH3, 1 mmol) of different chain lengths, and stirred at room temperature in the dark for 12 h. After the reaction was complete, the reaction solution was washed once with 100 mL of dilute hydrochloric acid and three times with distilled water. The organic layer was dried over anhydrous magnesium sulfate and filtered. The crude product was separated by thin-layer chromatography using ethyl acetate and ethanol in a 5:1 ratio to obtain a blue-black solid product HB-8-PEGn-CH3 (n = 2, 8, 16, 32).

[0222] Preparation of HC derivatives of ethanolamine with different chain lengths of polyethylene glycol (R1 = -H, R2 = -NH-(CH2CH2-O)2-CO-PEGn-CH3) (n = 2, 8, 16, 32): The above experiments were repeated by replacing HB with HC to obtain the blue-black solid product HC-8-PEGn-CH3 (n = 2, 8, 16, 32). The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0223]

[0224] Example 29

[0225] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-CO-PEGn-H): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, and HOOC-PEGn-H as the polyethylene glycol, similar to the synthesis method of HB-8-PEGn-CH3, the bamboo red pigment derivatives HB-8-PEGn-H (n = 2, 16, 32) and deacetylated bamboo red pigment derivatives HC-8-PEGn-H (n = 8, 16, 32) were obtained. The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0226]

[0227] Example 30

[0228] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-CO-PEGn-CH2CH2CO2CH3): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, and HOOC-PEGn-CH2CH2CO2CH3 as polyethylene glycol, similar to the synthesis method of HB-8-PEGn-CH3, the bamboo red pigment derivatives HB-8-PEGn-CO2CH3 (n = 2, 8, 32) and deacetylated bamboo red pigment derivatives HC-8-PEGn-CO2CH3 (n = 8, 16, 32) were obtained. The characterization results of the above derivatives are shown in Table 1, and the synthetic route is shown below:

[0229]

[0230] Example 31

[0231] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-CO-PEGn-CH2CH2SO3H): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, and HOOC-PEGn-CH2CH2SO3H as the polyethylene glycol, the synthesis method was similar to the HB-8-PEGn-CH3 synthesis method in Example 28, yielding the bamboo red pigment derivative HB-8-PEGn-SO3H (n = 2, 8, 16) and the deacetylated bamboo red pigment derivative HC-8-PEGn-SO3H (n = 2, 8, 16). The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0232]

[0233] Example 32

[0234] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-COCH2-PEGn-CH3): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, and HOOC-CH2-PEGn-OCH3 polyethylene glycol as the raw material, similar to the synthesis method of HB-8-PEGn-CH3, the bamboo red pigment derivative HB-9-PEGn-CH3 (n = 2, 8, 16) and the deacetylated bamboo red pigment derivative HC-9-PEGn-CH3 (n = 2, 8, 16) were obtained. The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0235]

[0236] Example 33

[0237] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-COCH2-PEGn-H): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, and HOOC-CH2-PEGn-H as the polyethylene glycol, similar to the synthesis method of HB-8-PEGn-CH3, bamboo red pigment derivatives HB-9-PEGn-H (n = 2, 8, 16) or deacetylated bamboo red pigment derivatives HC-9-PEGn-H (n = 2, 8, 16) were obtained. The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0238]

[0239] Example 34

[0240] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-COCH2-O-PEGn-CH3): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, and HOOC-CH2-O-PEGn-CH3 polyethylene glycol as the raw material, similar to the synthesis method of HB-8-PEGn-CH3, bamboo red fungicide derivatives HB-10-PEGn-CH3 (n = 2, 8, 16) and deacetylated bamboo red fungicide derivatives HC-10-PEGn-CH3 (n = 2, 8, 16) were obtained. The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0241]

[0242] Example 35

[0243] Preparation of HB or HC derivatives with different chain lengths of glycoside diamine-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)2-COCH2-O-PEGn-H): Using HB-8 or HC-8 synthesized in Example 28 as raw materials, similar to the synthesis method of HB-8-PEGn-CH3, bamboo red sclerotin derivatives HB-10-PEGn-H (n = 2, 8, 16) or deacetylated bamboo red sclerotin derivatives HC-10-PEGn-H (n = 2, 8, 16) were obtained. The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0244]

[0245] Example 36

[0246] Preparation of HB or HC derivatives with different chain lengths of glycotriol amino-substituted polyethylene glycol (R1 = -COCH3 or -H, R2 = -NH-(CH2CH2-O)3-CO-PEGn-CH3): The amino substituent used was H2N-(CH2CH2-O)3-H, and the polyethylene glycol used was HOOC-PEGn-CH3. The synthesis method was similar to the synthesis method of HB-8-PEGn-CH3 in Example 28, yielding the bamboo red pigment derivative HB-11-PEGn (n = 8, 16, 24) and the deacetylated bamboo red pigment derivative HC-11-PEGn (n = 8, 16, 24). The characterization results of the above derivatives are shown in Table 1, and their synthetic routes are as follows:

[0247]

[0248] Example 37

[0249] 1) Absorption spectrum

[0250] Figure 4 The absorption spectra of the compounds related to this invention are shown. Figure 4 In (a), the commercially available porphyrin-based photosensitizer PpIX exhibits multi-band absorption, all of which are narrow absorption bands. The maximum absorption wavelengths suitable for phototherapy are 570 nm and 630 nm, and their molar extinction coefficients are both below 8000 M. -1 cm -1 The commercially available porphyrin-based photosensitizer Ce6 has a maximum absorption wavelength of 650 nm and a molar extinction coefficient of approximately 15,000 MΩ. -1 cm -1 Furthermore, commercially available PpIX and Ce6 exhibit narrow absorption within the phototherapy window, thus limiting their light absorption capacity within this window. However, the absorption spectra of the bamboo red fungin disclosed in this invention and its polyethylene glycol-modified product are entirely different. For example... Figure 4 As shown in (b), the maximum absorption peak of HB (a type of styrax spp.) is around 470 nm. The derivative HB-1-PEG8-CH3 prepared in Example 2 has a broad and strong absorption in the phototherapy window, with a wide absorption band between 500-750 nm. It forms a strong continuous absorption band in the phototherapy window, with a maximum absorption peak around 580 nm, which is about 110 nm redshifted from the maximum absorption peak of HB. The molar extinction coefficient is approximately 20000 M. -1 cm -1The absorption spectrum of the derivative HC-1-PEG8-CH3 prepared in Example 7 also falls within the ideal phototherapy window, exhibiting a broad absorption band between 500-750 nm and a maximum absorption peak around 580 nm, demonstrating strong red light absorption. The absorption spectra of other bamboo red pigment derivatives of this invention are similar to HB-1-PEG8-CH3, exhibiting a broad absorption band between 500-750 nm and a maximum absorption peak around 580 nm. Therefore, the absorption wavelength and light absorption capacity of the 2-position polyethylene glycol-substituted water-soluble bamboo red pigment derivatives of this invention are far superior to the commercial photosensitizers PpIX and Ce6, exhibiting a more prominent red light absorption capacity. The maximum absorption wavelengths of some compounds described in this invention are shown in Table 1.

[0251] 2) Reactive oxygen species

[0252] The reactive oxygen species of the derivatives described in this invention were determined by paramagnetic resonance (ESR). Figure 5 The ESR (Enhanced Reactive Oxygen Species) curve of the reactive oxygen species in the derivative HB-1-PEG16-CH3 prepared in Example 2 is shown. Experiments show that HB-1-PEG16-CH3 can efficiently generate reactive oxygen species (ROS). Measurements using singlet oxygen and superoxide radical scavengers indicate that this derivative can efficiently generate photosensitive reactive species, primarily singlet oxygen. Figure 5 a) can also produce a small amount of superoxide radicals ( Figure 5 (b) Both of these reactive oxygen species are beneficial for photodynamic therapy. The derivatives disclosed in other embodiments of this invention also possess the ability to efficiently generate singlet oxygen and assist in the generation of small amounts of superoxide radicals.

[0253] 3) Singlet oxygen yield

[0254] Singlet oxygen quantum yield test: Sodium 9,10-anthraquinone malonate (ADPA) was used as the singlet oxygen scavenger, and methylene blue (MB) was used as the standard photosensitizer (singlet oxygen yield 0.52). The ADPA solution was added to a solution of the derivative described in this invention or MB, and the sample was taken at 635 nm (0.1 W cm⁻¹). -2 Using laser as the light source, the absorption intensity of ADPA at 378 nm was plotted against irradiation time, and the singlet oxygen yield of the derivative was calculated. Figure 6 (a) and (b) show the photodegradation curves of derivatives HB-1-PEG8-CH3 (Example 2) and HC-1-PEG8-CH3 (Example 7), respectively. It can be seen that the photosensitizer significantly generates singlet oxygen under light irradiation, thereby significantly degrading ADPA. This is achieved through comparison and calculation of the singlet oxygen efficiency curve with the reference compound, Rose Red RB. Figure 6 (c) and Figure 6(d) The singlet oxygen efficiencies of HB-1-PEG8-CH3 and HC-1-PEG8-CH3 were 0.32 and 0.36, respectively. The bamboo red fungicide derivatives disclosed in other embodiments of this invention all exhibit singlet oxygen generation efficiencies between 0.2 and 0.4, demonstrating a high capacity for generating reactive oxygen species. The singlet oxygen yields of the compounds described in this invention are shown in Table 1.

[0255] 4) Water-soluble

[0256] The bamboo red mycotoxin derivatives disclosed in this invention all contain hydrophilic groups of polyethylene glycol, giving the photosensitizer molecules strong water solubility under physiological conditions. Experiments show that more than 10 mg of the photosensitizer described in this invention can be dissolved in each milliliter of physiological saline or glucose injection, exhibiting excellent water solubility. This allows the derivatives described in this invention to be well transported in blood vessels during intravenous injection without causing vascular blockage. HB-1-PEG8-CH3 (Example 2) contains 8 ethylene glycol units with methyl end groups, and is soluble in more than 10 mg per milliliter of physiological saline; HB-1-PEG18-H (Example 3) contains 18 ethylene glycol units with hydrogen end groups, and is soluble in more than 15 mg per milliliter of physiological saline; HC-1-PEG16-H (Example 8) contains 16 ethylene glycol units, and is soluble in more than 15 mg per milliliter of physiological saline; HB-8-PEG16-CH3 (Example 28) contains 16 ethylene glycol units with methyl end groups, and is soluble in more than 20 mg per milliliter of physiological saline; HC-8-PEG16-CO2CH3 (Example 30) contains 16 ethylene glycol units with ester end groups, and is soluble in more than 15 mg per milliliter of physiological saline; all these compounds exhibit excellent water solubility. The derivatives disclosed in other embodiments of the present invention also have good water solubility and biocompatibility, and can dissolve more than 3 to 20 mg of the derivatives of the present invention per milliliter of physiological saline.

[0257] 5) Light stability

[0258] The photostability comparison between the derivatives prepared in this invention and commercial photosensitizers is as follows: Figure 7 As shown. From Figure 7 As can be seen in (a), a 635nm laser at 30mW / cm² can achieve the desired effect. 2 After 30 minutes of illumination under high light intensity, the absorption spectrum of HB-1-PEG16-CH3 (Example 2) did not show a significant decrease, with the absorption intensity at its maximum wavelength decreasing by less than 5% (curve i). Under the same conditions, the absorption intensity at its maximum wavelength of HC-1-PEG16-CH3 (Example 7) also decreased by less than 10%, exhibiting similarly good photostability (curve ii). Meanwhile, under the same conditions, using a 635nm laser at 30mW / cm²... 2After 30 minutes of illumination under high light intensity, the maximum absorption of the commercially available porphyrin-based photosensitizer Ce6 decreased by 30% (curve iii); while the absorption spectrum of the commercially available hematoporphyrin-based photosensitizer HpD decreased even more, reaching approximately 50% (curve iv). Figure 7 As shown in (b), under the same conditions, the absorption intensity at the maximum wavelength of HB-8-PEG8-CH3 (Example 28) decreased by less than 10%, exhibiting good photostability (curve i); the absorption intensity at the maximum wavelength of HC-8-PEG8-CH3 (Example 28) also decreased by less than 10%, similarly exhibiting good photostability (curve ii). The derivatives disclosed in other embodiments of this invention also exhibit good photostability, with their absorption intensity at the maximum wavelength decreasing by approximately less than 10% under the same conditions. Therefore, the bamboo red mycotoxin derivatives prepared in this invention have better photostability than commercial photosensitizers.

[0259] 6) pH stability

[0260] 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 spectrum of these derivatives does not change significantly within the pH range of 6.2–8.0, indicating that these derivatives have good pH stability under physiological conditions. Figure 8 As shown in (a), curve (ii) shows that the absorption spectrum of HB-1-PEG24-CO2CH3 (Example 5) does not change significantly in the pH range of 6.2 to 8.0; curve (iii) shows that the absorption spectrum of HC-1-PEG24-CO2CH3 (Example 10) does not change significantly in the pH range of 6.2 to 8.0. It exhibits good pH stability within this range because the two phenolic hydroxyl groups of bamboo red fungicide are not easily deprotonated under these acidic and alkaline conditions. In contrast, commercially available HpD contains two carboxyl groups, which can be deprotonated in the pH range of 6.2 to 8.0, resulting in a significant change in the absorption spectrum and thus demonstrating the instability of the HpD photosensitizer (curve i). Similarly, as... Figure 8 As shown in (b), curve (ii) indicates that the absorption spectrum of HB-8-PEG16-H (Example 29) shows no significant change in the pH range of 6.2–8.0; curve (iii) shows the maximum absorption curve of HC-8-PEG16-H (Example 29) as a function of pH, indicating good pH stability within this range. This is also because the two phenolic hydroxyl groups of bamboo red fungicide are less prone to deprotonation under these acidic and alkaline conditions. The derivatives disclosed in other embodiments of the present invention also exhibit good pH stability under physiological conditions, with little impact on the absorption spectrum of the photosensitizing drug from pH changes in the range of 6.2–8.0, demonstrating better pH stability than commercially available HpD.

[0261] 7) Photothermal conversion efficiency

[0262] The derivative of the present invention was subjected to laser light of different intensities (0.1–0.8 W / cm²) at a wavelength of 635 nm. -2 The temperature change was detected under irradiation. Photothermal conversion efficiency test: Solutions of different concentrations of derivatives (0-50 μM, 1.0 mL) were added to quartz cuvettes, and photothermal performance was measured under laser irradiation of a specific wavelength and intensity. Temperature changes were recorded using thermocouples and a digital thermometer, and real-time thermal imaging of the samples was recorded using a Fotric220 infrared thermal imager. The solution of bamboo red mycotoxin was irradiated with a 635 nm laser (0.8 W / cm²). -2 Irradiation was performed for 15 minutes followed by natural cooling, and the photothermal conversion efficiency was measured. The photothermal conversion efficiency η was calculated using the formula:

[0263]

[0264] Where h is the heat transfer coefficient, A is the surface area of ​​the quartz cell, and T is the heat transfer coefficient. max This is the highest temperature at which the derivative solution described in this invention is heated, T. surr It is room temperature, Q dis The heat loss is caused by the absorption of light by the quartz sample cell and solvent; I is the laser power; A is the heat loss caused by the absorption of light by the quartz sample cell and solvent. 635 It is the absorbance of the bamboo red mycotoxin derivative described in this invention at 635 nm.

[0265] The bamboo red mycin derivative prepared by this invention exhibits good photothermal conversion efficiency under 635nm laser irradiation.

[0266] Derivative HB-1-PEG16-CH3 (Example 2) under different laser intensities (0.1–0.8 W cm⁻¹) -2 The photothermal change curve under irradiation is as follows: Figure 9 As shown in (a). When the laser intensity is 0.1 W / cm². -2 When irradiating the HB-1-PEG16-CH3 solution (concentration 10 μM) for 10 min, the temperature increased by only 3.0 °C, and the photothermal effect was negligible; when the light intensity was increased to 0.5 W cm⁻¹… -2 (All other experimental conditions remained the same), the solution temperature increased by 18.5℃; when the light intensity continued to increase to 0.8 W cm⁻¹ -2 At that time, the solution temperature increased by 28.8℃, producing a significant photothermal effect. This indicates that the HB-1-PEG16-CH3 solution under laser irradiation can simultaneously generate singlet oxygen and photothermal effects, and can be used for photodynamic / photothermal synergistic therapy.

[0267] The derivative prepared in this invention has a wavelength of 635 nm and a wavelength of 0.8 W / cm². -2 The photothermal change curve under laser irradiation is as follows: Figure 9As shown in (b). Using pure water as a blank control group, the temperature of the pure water only increased by 4.5℃; the derivative HB-1-PEG16-CH3 prepared in Example 2 was heated at 0.8W cm⁻¹. -2 Under laser irradiation, the solution temperature increased by 28.8℃, producing a significant photothermal effect with a photothermal conversion efficiency of 35%. The HC-1-PEG16-CH3 prepared in Example 7 was subjected to 0.8W cm⁻¹ irradiation. -2 Under laser irradiation, the solution temperature increased by 25.8℃, producing a significant photothermal effect with a photothermal conversion efficiency of 33%. The HB-1-PEG12-H prepared in Example 3 was subjected to 0.8W cm⁻¹ irradiation. -2 Under laser irradiation, the solution temperature increased by 23.5℃, producing a significant photothermal effect with a photothermal conversion efficiency of 31%. Under the same conditions, the HC-1-PEG12-H prepared in Example 8 showed a 22.4℃ increase in solution temperature, also producing a significant photothermal effect with a photothermal conversion efficiency of 30.8%. Besides the above derivatives, other derivatives of this invention also exhibit good photothermal conversion performance, with photothermal conversion efficiencies ranging from 25% to 35%. Specific photothermal conversion efficiencies are shown in Table 1.

[0268] Table 1: Characterization data of the bamboo red mycotoxin derivatives prepared in Examples 2-36 of the present invention.

[0269]

[0270]

[0271]

[0272] Example 38 Cell fluorescence imaging

[0273] Cell fluorescence imaging of the bamboo red fungin derivative prepared in this invention is as follows: Figure 10 As shown. Confocal fluorescence imaging results indicate that HB-1-PEG16-CH3 (Example 2) has good water solubility and biocompatibility. When incubated with glioma cells (C6 cells), the photosensitizer was found to rapidly enter the lysosomes of C6 cells and produce good red fluorescence imaging in the cells. This indicates that HB-1-PEG16-CH3 can be used for fluorescence imaging of glioma cells, and the accumulation, distribution and metabolism of the drug in vivo can be tracked through fluorescence detection.

[0274] Besides glioma C6 cells, HB-1-PEG16-CH3 can also enter the lysosomes of various other cell types, such as esophageal cancer cells (AKR), gastric cancer cells (MFC), lung cancer cells (A549), liver cancer cells (HCC), bile duct cancer cells (MCC), colon cancer cells (HCT116), head and neck cancer cells (SCC2), brain cancer cells (G442), tongue cancer cells (TSCCa), nasal cancer cells (KB), oral cancer cells (CAL27), basal cell carcinoma cells (BCC), squamous cell carcinoma cells (PECA), melanoma cells (B16), prostate cancer cells (LNCaP), and bladder cancer cells (MBT-2). Using DCFH-DA to detect intracellular singlet oxygen, the photosensitizer and fluorescent probe DCFH-DA were co-incubated intracellularly. With the irradiation time increasing to 120 s, the green fluorescence intensity gradually increased, indicating an increase in intracellular singlet oxygen. Other derivatives of this invention can also perform excellent cell fluorescence imaging, allowing for the tracking of drug accumulation, distribution, and metabolism in vivo through fluorescence detection.

[0275] Example 39 Cytotoxicity

[0276] Dark cytotoxicity and phototoxicity experiments were performed on the following tumor cells: 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) associated with gastrointestinal tumors; head and neck cancer cells (SCC2), brain cancer cells (G442), tongue cancer cells (TSCCa), nasal cancer cells (KB), oral cancer cells (CAL27), and glioma cells (C6) associated with head, neck, and facial tumors; basal cell carcinoma cells (BCC), squamous cell carcinoma cells (PECA), and melanoma cells (B16) associated with skin tumors; and prostate cancer cells (LNCaP) and bladder cancer cells (MBT-2) associated with the reproductive and urinary systems.

[0277] 1) Cell dark toxicity assay:

[0278] The cultured tumor cells were added to a culture medium (containing serum proteins and antibiotics), digested with 0.25% trypsin, and pipetted to prepare a single-cell suspension. The cell count was adjusted to approximately 2 x 102. 4Cells were seeded at a density of 200 μL / well in a 96-well plate and incubated at 37°C in a 5% CO2 incubator. After cell attachment, the supernatant was discarded, and different concentrations of photosensitizer (commercial HpD, the derivative described in this invention) were added under light-protected conditions according to the experimental design. Cells were incubated for another 1 hour, and cell viability was assessed using the MTT assay. Alternatively, 20 μL of MTT (tetramethyl azo blue) was added to each well, and the plate was incubated for another 4 hours. Then, 150 μL of dimethyl sulfoxide was added to each well, and the plate was shaken for 10 minutes to dissolve the purple crystals. The optical density (OD) value of each well was measured at 570 nm using a microplate reader, and cell viability was calculated as: Cell viability = (OD value of experimental group / OD value of blank group) × 100%.

[0279] like Figure 11 As shown in (a), the cytotoxicity (dark toxicity) study showed that the HB-1-PEG8-CH3 synthesized in Example 2 had low cytotoxicity, similar to the commercial photosensitizing drug hematoporphyrin HpD. Esophageal cancer cells AKR were incubated with 10 μM of the photosensitizing drug for half an hour, and no obvious death of AKR cells was observed, indicating that this type of photosensitizing drug has basically no cytotoxicity.

[0280] The dark toxicity results of some derivatives of this invention are as follows: Figures 11-15 As shown in Tables 2-4, the bamboo red mycotoxin derivatives described in this invention have virtually no cytotoxicity, and cell viability is above 90% within a concentration range of 20 μM.

[0281] 2) Cell phototoxicity assay (0.1 W / cm) 2 ):

[0282] The phototoxicity assay was performed in the same manner as the dark toxicity assay, except that the incubated cells were irradiated with a 635nm semiconductor laser at a power density of 0.1W / cm². 2 (At this light intensity, only photodynamic effects occur, not photothermal effects), ensuring the light beam is uniformly and perpendicularly irradiated onto the 96-well culture plate for 1000 seconds. Figure 11 (b) The phototoxicity study showed that HB-1-PEG8-CH3 exhibited a very strong killing effect on esophageal cancer cells (AKR) under red light irradiation. A concentration of 100 nM was sufficient to kill over 90% of AKR cells, while under the same conditions, the commercially available photosensitizer HpD could only kill about 20% of AKR cells. This indicates that the photodynamic effect of this derivative is significantly superior to that of the commercially available photosensitizer HpD.

[0283] The phototoxicity test results of some derivatives in this invention are as follows: Figures 11-15As shown in Tables 2-4, the bamboo red fungicide derivative of this invention can kill more than 80-90% of tumor cells at a concentration of 50 nM, and its half-lethal concentration (IC50) is [not specified]. 50 The value is approximately 20–30 nM. Therefore, the derivative disclosed in this invention exhibits better photodynamic effects than the commercial photosensitizer HpD.

[0284] Table 2: MTT data of the bamboo red fungus derivative of the present invention on gastrointestinal tumor cells (0.1 W / cm²). 2 )

[0285]

[0286]

[0287] Table 3: MTT data of some bamboo red mycotoxin derivatives of the present invention against head, neck and facial tumor cells (0.1 W / cm²). 2 )

[0288]

[0289]

[0290] Table 4: MTT data of some bamboo red mycotoxin derivatives of the present invention against skin tumor and genitourinary tumor cells (0.1 W / cm²). 2 )

[0291]

[0292] Figure 11 Dark toxicity (a) and phototoxicity (b) of HB-1-PEG8-CH3 prepared in Example 2 on esophageal cancer AKR cells and HB-1-PEG16-CH3 on gastric cancer MFC cells. Figure 11 As shown in (a), the cytotoxicity (dark toxicity) of the photosensitizers indicates that HB-1-PEG8-CH3, whether with 8 or 16 ethylene glycol units, has low cytotoxicity to esophageal cancer AKR cells, and HB-1-PEG16-CH3 has low cytotoxicity to gastric cancer MFC cells. Similar to the commercially available photosensitizer hematoporphyrin HpD, no significant cell death was observed when AKR cells were incubated with 20 μM HB-1-PEG8-CH3 for half an hour, or when MFC cells were incubated with 20 μM HB-1-PEG16-CH3 for half an hour, indicating that this type of photosensitizer has virtually no cytotoxicity. Figure 11 (b) indicates that the photosensitizing drug exhibits very strong killing effect on esophageal cancer AKR and gastric cancer MFC cells under red light irradiation. A concentration of 50 nM HB-1-PEG8-CH3 can kill more than 85% of AKR cells, with a median lethal concentration (IC50) of 1 / 3. 50The median lethal concentration (LC50) of HB-1-PEG16-CH3 is approximately 25 nM; similarly, 50 nM of HB-1-PEG16-CH3 can kill more than 90% of gastric cancer MFC cells, with an IC50 value of approximately 25 nM. 50 The value is approximately 25 nM; while under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 20% of AKR and MFC cells, indicating that the photodynamic effect of this type of hematoporphyrin polyethylene glycol derivative is significantly better than that of the commercial photosensitizer hematoporphyrin HpD.

[0293] Figure 12 The images show the dark toxicity (a) and phototoxicity (b) of the tranexamic acid-substituted deacetylated bamboo red cinnamic acid product HC-1-PEG8-CH3 prepared in Example 7 against lung cancer cells A549 and liver cancer HCC cells, respectively. Figure 12 As shown in (a), cytotoxicity (dark toxicity) indicates that HC-1-PEG8-CH3 and HC-1-PEG16-CH3 have low cytotoxicity, similar to the commercial photosensitizer hematoporphyrin HpD. When A549 cells or HCC cells were co-incubated with 20 μM HC-1-PEG8-CH3 and 20 μM HC-1-PEG16-CH3 for half an hour, no significant cell death was observed in A549 and HCC cells, indicating that such photosensitizers are basically non-cytotoxic. Figure 12 (b) indicates that under 635 nm laser irradiation, the photosensitizing drug exhibited very strong killing effects on A549 and HCC cells, respectively. A concentration of 50 nM HC-1-PEG8-CH3 could kill more than 80% of A549 cells, with a median lethal concentration (IC50) of [missing value]. 50 The value is approximately 30 nM; similarly, a concentration of 50 nM HC-1-PEG16-CH3 can kill more than 90% of HCC cells, with an IC50 (half-lethal concentration) of 30 nM. 50 The value is approximately 30 nM; while under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 25% of A549 and HCC cells, indicating that the photodynamic effect of this type of polyethylene glycol derivative of hematoporphyrin is significantly better than that of the commercial photosensitizer hematoporphyrin HpD.

[0294] Figure 13 The diagram shows the dark toxicity (a) and phototoxicity (b) of the bamboo red sclerotin and glycoside diamine-polyethylene glycol derivative HB-8-PEG8-CH3 prepared in Example 28 against cholangiocarcinoma MCC cells, and the dark toxicity (a) and phototoxicity (b) of HB-8-PEG16-CH3 against colon cancer HCT116 cells. It can be seen that HB-8-PEG8-CH3 and HB-8-PEG16-CH3 containing long-chain PEG have almost no cytotoxicity in the absence of light. After irradiation with 635 nm light, a concentration of 50 nM can kill more than 90% of cholangiocarcinoma MCC cells, with a median lethal concentration (IC50) of 1 / 3.50 The value is approximately 30 nM; a concentration range of 50 nM can kill more than 90% of HCT116 in colon cancer, with an IC50 (median lethal concentration). 50 The value is approximately 30 nM; while under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 25% of MCC and HCT116 cells, which also indicates that the photodynamic effects of HB-8-PEG8-CH3 and HB-8-PEG16-CH3 are significantly better than those of the commercial photosensitizer hematoporphyrin HpD.

[0295] Figure 14 (a) Phototoxicity diagrams of HC-8-PEG2-CH3 prepared in Example 28 against G442 brain cancer cells and SCC2 head and neck cancer cells. The photosensitizing drug exhibited very strong killing activity against G442 or SCC2 cells under 635 nm laser irradiation. A concentration of 50 nM HC-8-PEG2-CH3 could kill more than 80% of G442 cells, with a median lethal concentration (IC50) of 100%. 50 The value was approximately 20 nM; similarly, a concentration of 50 nM HC-8-PEG8-CH3 could kill more than 90% of SCC2 cells, with an IC50 (half-lethal concentration) of 20 nM. 50 The value is approximately 20 nM; while under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 20% of brain cancer cells G442 and head and neck cancer cells SCC2.

[0296] Figure 14 (b) Phototoxicity graphs of compound HC-8-PEG16-H (prepared in Example 29) against tongue cancer cells TSCCa and HC-8-PEG32-H against nasal cancer cells KB. A concentration of 50 nM HC-8-PEG16-H can kill more than 90% of tongue cancer cells TSCCa, with a median lethal concentration (IC50) of 100%. 50 The value is approximately 30 nM; while a concentration of 50 nM HC-8-PEG32-H can kill more than 90% of nasopharyngeal carcinoma cells (KB), with an IC50 (half-lethal concentration). 50 The value is approximately 30 nM; while under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 20% of tongue cancer cells (TSCCa) or nasal cancer cells (KB).

[0297] Figure 14 (c) Phototoxicity diagrams of the compound HC-8-PEG2-SO3H prepared in Example 31 of this invention against oral cancer cells CAL27 and against glioma cells C6. A concentration of 50 nM HC-8-PEG2-SO3H can kill more than 90% of oral cancer cells CAL27, with a median lethal concentration (IC50) of 1 / 3. 50The value is approximately 30 nM; while a concentration of 50 nM HC-8-PEG8-SO3H can kill more than 90% of glioma C6 cells, with an IC50 (median lethal concentration). 50 The photodynamic effect of the hematoporphyrin derivative HpD on oral cancer cells (CAL27) and glioma cells (C6) was approximately 30 nM. Under the same conditions, HpD only killed about 30% of these cells. These results indicate that this type of hematoporphyrin-based polyethylene glycol derivative has a significantly better photodynamic effect on oral cancer cells (CAL27) and glioma cells (C6) than the commercially available hematoporphyrin HpD.

[0298] Figure 15 (a) Phototoxicity diagrams of HB-1-PEG4-H (Example 3), HB-1-PEG16-CO2H (Example 4), and HB-1-PEG24-CO2CH3 (Example 5) on basal cell carcinoma (BCC) cells. The diagrams show that under 635nm red light irradiation, a 50nM concentration of HB-1-PEG4-H can kill more than 80% of BCC cells; while under the same conditions, a 50nM concentration of HB-1-PEG16-CO2H can kill more than 80% of BCC cells, and a 50nM concentration of HB-1-PEG24-CO2CH3 can kill more than 85% of BCC cells. The median lethal concentration (IC50) is [not specified in the original text]. 50 The values ​​were all around 20 nM, indicating that the three photosensitizers mentioned above have a good ability to photodynamically inactivate tumor cells in basal cell carcinoma (BCC) cells.

[0299] Figure 15 (b) Phototoxicity diagrams of HC-1-PEG2-H (Example 8), HC-1-PEG24-SO3H (Example 11), and HB-1-NH-PEG4-CH3 (Example 12) on melanoma B16 cells. The diagrams show that under 635nm red light irradiation, HC-1-PEG2-H at a concentration of 50 nM can kill more than 80% of B16 cells; while under the same conditions, HC-1-PEG24-SO3H at a concentration of 50 nM and HB-1-NH-PEG4-CH3 at a concentration of 50 nM can kill more than 85% of B16 cells, and their median lethal concentration (IC50) is [not specified]. 50 The values ​​were all around 30 nM, indicating that the three photosensitizers mentioned above have a good ability to photodynamically inactivate tumor cells in melanoma B16 cells.

[0300] Figure 15(c) Phototoxicity diagrams of HB-1-NH-PEG16-H (Example 13), HC-1-NH-PEG4-CH3 (Example 17), and HC-1-NH-PEG24-CO2CH3 (Example 20) on squamous cell carcinoma PECA cells. The diagrams show that under 635nm red light irradiation, a 50nM concentration of HB-1-NH-PEG16-H can kill over 90% of PECA cells; while under the same conditions, a 50nM concentration of HC-1-NH-PEG4-CH3 can kill over 85% of PECA cells, and a 50nM concentration of HC-1-NH-PEG24-CO2CH3 can kill over 90% of PECA cells. The median lethal concentration (IC50) is [not specified in the original text]. 50 The value is approximately 20 nM, indicating that the three photosensitizers mentioned above have a good ability to photodynamically inactivate tumor cells in squamous cell carcinoma PECA cells.

[0301] Figure 15 (d) Phototoxicity diagrams of HB-2-p-PEG4 in Example 22, HB-5-PEG10 in Example 25, and HB-6-PEG24 in Example 26 on prostate cancer LNCaP cells; The diagrams show that under 635nm red light irradiation, a concentration of 50nM HB-2-p-PEG4 can kill more than 85% of LNCaP cells; while under the same conditions, a concentration of 50nM HB-5-PEG10 can kill more than 85% of LNCaP cells, and a concentration of 50nM HB-6-PEG24 can kill more than 90% of LNCaP cells, with a median lethal concentration (IC50) of 90%. 50 The values ​​were all around 20 nM, indicating that the three photosensitizers mentioned above have a good ability to photodynamically inactivate tumor cells in prostate cancer LNCaP cells.

[0302] Figure 15 (e) Phototoxicity diagrams of HB-7-PEG4, HC-7-PEG16 in Example 27, and HB-8-PEG32-CO2CH3 in Example 30 on bladder cancer MBT-2 cells. The diagrams show that under 635nm red light irradiation, HB-7-PEG4 at a concentration of 50 nM can kill more than 90% of MBT-2 cells; under the same conditions, HC-7-PEG16 at a concentration of 50 nM can kill more than 90% of MBT-2 cells, and HB-8-PEG32-CO2CH3 at a concentration of 50 nM can kill more than 90% of MBT-2 cells. The median lethal concentration (IC50) is [not specified in the original text]. 50 The values ​​were all around 20 nM, indicating that the three photosensitizers mentioned above have a good ability to photodynamically inactivate tumor cells in bladder cancer MBT-2 cells.

[0303] Figure 15 (f) Phototoxicity diagrams of HB-9-PEG16-CH3 in Example 32, HB-10-PEG2-CH3 in Example 34, and HB-11-PEG16 in Example 36 on basal cell carcinoma (BCC) cells. The diagram shows that under 635nm red light irradiation, 50nM HB-9-PEG16-CH3 killed more than 80% of BCC cells; 50nM HB-10-PEG2-CH3 killed more than 80% of BCC cells; and 50nM HB-11-PEG16 killed more than 85% of BCC cells. The median lethal concentration (IC50) of HB-9-PEG16 was [not specified in the original text]. 50 The values ​​were all around 30 nM, indicating that the three photosensitizers mentioned above have a good ability to photodynamically inactivate tumor cells in basal cell carcinoma (BCC).

[0304] In summary, the bamboo red mycotoxin derivative described in this invention exhibits almost no cytotoxicity in the absence of light, and demonstrates significant photodynamic effects under light conditions, especially under laser irradiation (0.1 W / cm²). 2 It exhibits strong photodynamic effects; photosensitizing drugs at concentrations of 10–50 nM can kill various tumor cells (such as: esophageal cancer cells AKR, gastric cancer cells MFC, lung cancer cells A549, liver cancer cells HCC, bile duct cancer cells MCC, colon cancer cells HCT116, head and neck cancer cells SCC2, brain cancer cells G442, tongue cancer cells TSCCa, nasal cancer cells KB, oral cancer cells CAL27, glioma cells C6, basal cell carcinoma cells BCC, squamous cell carcinoma cells PECA, melanoma cells B16, prostate cancer cells LNCaP, and bladder cancer cells MBT-2), with a killing rate of over 85%–90% and a median lethal concentration (IC50). 50 The value is approximately 20–30 nM, which is higher than the IC50 of the commercial photosensitizer hematoporphyrin HpD under the same conditions. 50 The values ​​are 1 to 2 orders of magnitude lower. Therefore, the bamboo red mycotoxin derivative disclosed in this invention has better dark toxicity, phototoxicity, and photodynamic effects than the commercial photosensitizer hematoporphyrin HpD.

[0305] 3) Cell phototoxicity assay (0.5 W / cm) 2 ):

[0306] The foregoing results indicate that the bamboo red mycotoxin derivative prepared in this invention exhibits good photothermal conversion, with a conversion efficiency between 25% and 35% (Table 1). Under high-intensity laser light (0.5 W cm⁻¹), -2 Irradiation can significantly increase the temperature in the environment surrounding the photosensitizing drug, producing a significant photothermal effect. This indicates that photosensitizers can simultaneously produce singlet oxygen and photothermal effects under appropriate laser irradiation, and can be used for photodynamic / photothermal synergistic therapy.

[0307] In the cell phototoxicity experiment, the incubated cells were irradiated with a 635nm wavelength laser, with the power density adjusted to 0.5W / cm². 2 The light beam was uniformly and perpendicularly irradiated onto the 96-well culture plate for 1000 seconds. At this light intensity, both photodynamic and photothermal effects occurred; under these conditions, tumor cell killing was a synergistic process of photodynamic and photothermal action. Some derivatives in this invention were used at 0.5 W / cm². 2 The phototoxicity test results are shown in Table 5. For example, the derivative HB-1-PEG8-CH3 (Example 2) under red light irradiation (0.5 W / cm²) 2 It showed stronger killing power against AKR esophageal cancer cells. A concentration of 30 nM could kill more than 95% of AKR cells, with a half-lethal concentration (IC50) of approximately 10 nM; while at 0.1 W / cm²... 2 Under red light irradiation (all other conditions being equal), a concentration of 50 nM is required to kill more than 90% of AKR cells, with a median lethal concentration of approximately 25 nM. Therefore, the synergistic effect of photodynamic and photothermal action significantly outperforms photodynamic action alone in killing esophageal cancer tumor cells.

[0308] Similarly, the derivative HC-1-PEG16-CH3 (Example 7) under red light irradiation (0.5 W / cm²) 2 It showed stronger killing power against bile duct cancer cells (MCC). A concentration of 30 nM could kill more than 95% of MCC cells, with a half-lethal concentration of approximately 10 nM; while at 0.1 W / cm²... 2 Under red light irradiation (all other conditions being equal), a concentration of 50 nM is required to kill more than 90% of MCC cells, with a median lethal concentration of approximately 30 nM. This also demonstrates that the synergistic effect of photodynamic and photothermal action significantly outperforms photodynamic action alone in killing cholangiocarcinoma tumor cells.

[0309] For example: the derivative HB-1-NH-PEG8-CH3 (Example 12) under red light irradiation (0.5W / cm²) 2 It exhibits stronger killing efficacy against C6 glioma cells. A concentration of 30 nM can kill over 95% of C6 cells, with a median lethal concentration of approximately 10 nM; while at 0.1 W / cm²... 2 Under red light irradiation (all other conditions being equal), a concentration of 50 nM was required to kill more than 85% of C6 cells, with a median lethal concentration of approximately 30 nM. This also demonstrates that the synergistic effect of photodynamic and photothermal action significantly outperforms photodynamic action alone in killing glioma cells.

[0310] For example: the derivative HC-1-NH-PEG16-CH3 (Example 17) under red light irradiation (0.5W / cm²) 2It exhibits stronger killing efficacy against basal cell carcinoma (BCC) cells. A concentration of 30 nM can kill over 95% of BCC cells, with a median lethal concentration (LD50) of approximately 10 nM; while at 0.1 W / cm²... 2 Under red light irradiation (0.5W / cm) 2 (Under the same conditions), a concentration of 50 nM is required to kill more than 90% of BCC cells, with a half-lethal concentration of approximately 25 nM. This also demonstrates that the synergistic effect of photodynamic and photothermal action significantly outperforms photodynamic action alone in killing basal cell carcinoma cells.

[0311] The above results demonstrate that the bamboo red mycotoxin derivative prepared in this invention possesses excellent photothermal conversion efficiency. Under high-intensity laser light (0.5W cm⁻¹), -2 Irradiation with lasers can significantly increase the temperature in the environment surrounding the photosensitizing drug, producing a noticeable photothermal effect. This indicates that photosensitizers can simultaneously generate singlet oxygen and photothermal effects under appropriate laser irradiation, making them suitable for photodynamic / photothermal synergistic therapy. Table 5 also lists other derivatives at 0.5 W / cm². 2 MTT assay results for killing various tumor cells under light irradiation. It can be seen that this type of derivative can kill more than 90-95% of tumor cells at a concentration of 30 nM, with a median lethal concentration (IC50) of [missing value]. 50 The value is approximately 10–15 nM. Therefore, the derivative described in this invention, under the synergistic effect of photodynamic and photothermal action, is significantly more effective in killing various tumor cells than photodynamic action alone.

[0312] Table 5: MTT data (0.5 W / cm²) of the derivatives described in this invention against gastrointestinal tumors, head and neck tumors, skin tumors, and genitourinary tumor cells. 2 )

[0313]

[0314]

[0315] Example 40: In vivo enrichment of photosensitizing drugs

[0316] To investigate the accumulation process of the bamboo red fungin derivative prepared in this invention in subcutaneous tumors, mice were used as a model. Esophageal cancer AKR cells were inoculated into subcutaneous tumors to obtain a mouse model of esophageal cancer subcutaneous tumors. The derivative HB-1-PEG16-CH3 from Example 2 of this invention was used as a photosensitizing drug and injected into tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in esophageal cancer AKR cell-bearing mice was observed. Fluorescence signals at the tumor site were collected after 4 hours using a multispectral small animal in vivo imaging system. The animal fluorescence imaging is shown below. Figure 17As shown in (a), it can be seen that after several cycles in vivo following intravenous injection, the drug molecule HB-1-PEG16-CH3 exhibits a strong accumulation capacity at the tumor site through passive targeting for approximately 4 hours.

[0317] A lung cancer A549 cells were then inoculated into subcutaneous tumors to obtain a subcutaneous mouse model of lung cancer. The derivative HC-1-PEG16-CH3 from Example 7 of this invention was used as a photosensitizing drug and injected into tumor-bearing mice via the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in lung cancer A549 cell-bearing mice was observed. Fluorescence signals at the tumor site were collected at 4 hours using a multispectral small animal in vivo imaging system. The animal fluorescence imaging is shown below. Figure 17 As shown in (b), it can be seen that after several cycles in vivo following intravenous injection, the drug molecule HC-1-PEG16-CH3 exhibits a strong accumulation capacity at the tumor site through passive targeting for approximately 4 hours.

[0318] The above results demonstrate 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 the small animals shows that these drugs produce significant fluorescent signals at the tumor sites in the mice, indicating that these derivatives can be used as photosensitizing drugs with good accumulation effects at the tumor sites in tumor-bearing mice. 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, prostate cancer, and bladder cancer.

[0319] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A bamboo red fungicide derivative of formula (I), and its isotopic label or pharmaceutically acceptable salt: Equation (I); In formula (I), the substituent R1 is -H or -COCH3; The general structural formula of substituent R2 is —NH—(CH2). p —X—Y—(CH2CH2O) n —Z; Equation (II); in, X is a linking group, selected from -(CH2CH2O). m -; Y is a linking group, selected from the following groups: -C(=O)-, -C(=O)-CH2- or -C(=O)-CH2O-, with X connected to the left side of the above group and polyethylene glycol unit connected to the right side; Z is a terminal base, selected from -H and -C. 1-6 Alkyl, -C 1-6 Alkyl-COOH, -C 1-6 Alkyl-COO-C 1-6 Alkyl, -C 1-6 Alkyl-SO3H, -C 1-6 Alkyl-SO3-C 1-6 alkyl; p is selected from 0 or 1; m is selected from 2 or 3; -(CH2CH2O) n - represents a polyethylene glycol unit, and n is an integer selected from 1 to 100.

2. The derivative according to claim 1, and its isotopic label or pharmaceutically acceptable salt, characterized in that, Z is selected from -H, -CH3, -C2H5, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H.

3. The derivative according to claim 1, and its isotopic label or pharmaceutically acceptable salt, characterized in that, R2 is selected from: -NH-(CH2CH2O) m -CO-(CH2CH2O) n -Z, -NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -Z -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -Z; In the above groups, m is selected from 2 or 3; each n is the same or different and is independently selected from integers from 1 to 100; Z is selected from -H, -CH3, -C2H5, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H.

4. The derivative according to claim 1, and its isotopic label or pharmaceutically acceptable salt, characterized in that, R2 is selected from: -NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH3, -NH-(CH2CH2O) m -CO-(CH2CH2O) n -H -NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2COOH、-NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2CH2COOH、-NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2CH2COOCH3、-NH-(CH2CH2O) m -CO-(CH2CH2O) n -CH2CH2SO3H、-NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -CH3、-NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -H、 -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -CH3 or -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -H; Where m is selected from 2 or 3, and each n is the same or different, and each is selected independently from an integer from 1 to 100.

5. The derivative according to claim 1, and its isotopic label or pharmaceutically acceptable salt, characterized in that, R2 is selected from: -NH-(CH2CH2O)2-CO-(CH2CH2O)2-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)4-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)5-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)6-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)7-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)8-CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O)9-CH3、-NH-(CH2CH2O)2-CO-(CH2CH2O) 10 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 12 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 14 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 16 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 18 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 20 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 24 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 32 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 36 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 40 -CH3, -NH-(CH2CH2O)2-CO-(CH2CH2O) 50 -CH3 or -NH-(CH2CH2O)2-CO-(CH2CH2O) 100 -CH3.

6. The derivative according to claim 1, and its isotopic label or pharmaceutically acceptable salt, characterized in that, 。 7. The derivative according to claim 1, and its isotopic label or pharmaceutically acceptable salt, characterized in that, The general structural formula of the bamboo red fungicide derivative described in formula (I) also includes the enol tautomer shown in formula (I'): 。 8. A pharmaceutical composition comprising at least one of the following: a bamboo red fungus derivative according to any one of claims 1-7, an isotopic label thereof, or a pharmaceutically acceptable salt thereof; said pharmaceutical composition as a photodynamic therapy drug and / or a fluorescence-mediated therapy drug and / or a photothermal therapy drug.

9. Use of at least one of the bamboo red fungin derivatives according to any one of claims 1-7 in the preparation of an antitumor drug, including its isotopic label or pharmaceutically acceptable salt.

10. The use according to claim 9, characterized in that, The drug is used as a photodynamic therapy drug and / or a fluorescence-mediated drug and / or a photothermal therapy drug.

11. The use according to claim 9, characterized in that, The tumors are gastrointestinal tumors, head and neck tumors, skin tumors, genitourinary system tumors, lung cancer, and liver cancer; the gastrointestinal tumors include esophageal cancer, gastric cancer, bile duct cancer, and colon cancer; the head and neck tumors include head and neck cancer, brain cancer, tongue cancer, nasal cancer, and oral cancer; the skin tumors include basal cell carcinoma, squamous cell carcinoma, and melanoma; and the genitourinary system tumors include prostate cancer and bladder cancer.

12. The use according to claim 11, characterized in that, The head, neck, and facial tumors include gliomas.

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