A water-soluble derivative of hypocrellin substituted with polyethylene glycol at the 2-position and its application in treating tumors

Abstract

The present invention discloses a water-soluble polyethylene glycol-modified hypocrellin derivative and its application in photodynamic therapy for tumors. The polyethylene glycol-modified hypocrellin derivative prepared by the present invention has a maximum absorption wavelength of about 580nm, a molar extinction coefficient of 15000-20000M-1cm-1, and has a strong light absorption capacity in the phototherapy window. Reactive oxygen species such as singlet oxygen can be efficiently produced under photosensitizing conditions, and a variety of tumor cells can be efficiently inactivated. The polyethylene glycol-modified hypocrellin derivative prepared by the present invention has good water solubility, and its solubility in physiological saline is above 10mg / mL. Such derivatives can be administered into the body through the tail vein as a photosensitizer, can specifically accumulate in tumor cells or tissues, and can be used for photodynamic drugs to treat the above-mentioned various tumors. Under the same conditions, the polyethylene glycol-substituted hypocrellin derivative involved in the present invention has a higher photodynamic therapy effect as a photosensitizer for treating tumors than porphyrin commercial photosensitizers.

Description

Technical Field

[0001] The invention relates to the technical field of photosensitizer drugs, and more specifically to a water-soluble polyethylene glycol-modified hypocrellin derivative, a preparation method thereof, and an application thereof in treating tumors. Background Art

[0002] Photodynamic therapy (PDT) is a new method for treating tumors that appeared in the late 1970s. It has now been officially approved in many countries such as the United States, Europe, Japan, and China, becoming a new technology for treating tumors. Photodynamic therapy is the ingestion of photosensitizers in the body of an organism. Within a certain period of time, photosensitizers can be more enriched in tumor cells and tissues. The tumor lesions are irradiated with a light source of the corresponding wavelength to stimulate the photosensitizer to produce reactive oxygen species. The reactive oxygen species produced (such as singlet oxygen, hydroxyl free radicals, and superoxide free radicals) can damage tumor cells and destroy tumor tissues through photochemical reactions, thereby achieving the purpose of tumor treatment. Compared with traditional surgery, chemotherapy, and radiotherapy, photodynamic therapy is more targeted for the treatment of cancer, and can selectively eliminate primary and recurrent tumors and reduce damage to normal cells. PDT has been used to treat a variety of tumors and has a certain effect on controlling local growth of tumors. At present, photodynamic therapy has been reported in the treatment of surface 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 cancer, tongue cancer, oral cancer, etc.), brain tumors (glioma), reproductive tumors (prostate cancer, bladder cancer, cervical cancer, etc.) and digestive system tumors (choledochal cancer, gastric cancer, lung cancer, liver cancer, colorectal cancer, pancreatic cancer). PDT not only has a good therapeutic effect on many primary tumors, but also has unique technical advantages for many metastatic tumors. Studies have shown that photodynamic therapy directly kills tumor cells through the production of reactive oxygen species or blocks the blood vessels that maintain the growth of tumor cells. Direct killing of tumor cells utilizes the dual selectivity of photodynamic therapy, the selective retention of photosensitizers in tumor cells, and the generation of reactive oxygen species near the lesion under the irradiation of light waves of specific wavelengths, leading to the pathology and death of tumor cells. Anti-angiogenic photodynamic therapy is a method in which tumor cells rely on blood vessels for survival. Photodynamic therapy can damage the vascular system associated with the tumor, leading to ischemic death of the tumor. If light is applied during the peak concentration period of the photosensitizer in the blood vessels, it can cause microvascular damage and insufficient blood supply in the lesion, causing cell necrosis or apoptosis.

[0003] Photosensitizer, light source and oxygen are the three elements of photodynamic therapy. The key links of photodynamic therapy are that photosensitizers specifically gather at the target, light and photosensitizers produce chemical reactions to produce active substances such as singlet oxygen, and active substances damage target cells in a targeted manner. PDT has advantages over other treatment methods in the treatment of tumors. It can destroy tumor tissues, but it is basically non-toxic to normal tissues within the allowable range of light doses that destroy tumors, and has a therapeutic effect on the infiltrated edges of tumors. Photosensitizers used in photodynamic therapy for tumors need to meet the following conditions: 1) The chemical composition of the photosensitizer is single and has no obvious toxic effects on the human body; 2) It can target tumor cells and tissues and remain in tumor tissues, and rarely enter normal cells; 3) The photosensitizer has deep tumor tissue penetration for excitation light; 4) It can selectively damage tumor cells under the action of light, while causing less damage to normal cells and tissues; 5) It can be quickly excreted from the body after treatment without other toxic side effects. At present, the photosensitizers used in tumor research and treatment mainly include: hematoporphyrin derivatives (HpD), 5-aminolevulinic acid (5-ALA), talaporfin, temoporfin, verteporfin, etc. However, these photosensitizers all belong to porphyrin compounds, and 5-aminolevulinic acid is also converted into protoporphyrin through biosynthesis in the body. The main problem with porphyrin photosensitizers is that it is difficult to separate geometric isomers and it is difficult to obtain photosensitizers of a single component. The relatively complex components of these mixed photosensitizers are not conducive to the evaluation of later drug metabolism and toxicological analysis. In addition, porphyrin compounds have a long retention time in the body and require a long light avoidance period, which is not conducive to clinical use. In order to better play the role of photodynamic therapy in the treatment of tumors, it is necessary to develop new photosensitizers with high efficiency, low toxicity and stability with strong light absorption ability in the phototherapy window.

[0004] Hypocrellin belongs to the class of perylenequinone compounds and is a natural plant photosensitizer. It is produced by a parasitic fungus, Hypocrellin, on the arrow bamboo at an altitude of more than 3,000 meters on the Yunnan Plateau in my country. Natural Hypocrellin is mainly in the form of Hypocrellin A (HA) (95%), and a small amount exists in the form of Hypocrellin B (HB) (5%). Hypocrellin has strong absorption in the visible light region and a large molar extinction coefficient, and can efficiently produce reactive oxygen under photosensitivity conditions. Many studies in the past have shown that Hypocrellin has the advantages of good phototoxicity, low dark toxicity, clear structure, fast metabolism in the body, and no toxic side effects in the body. However, natural Hypocrellin is a hydrophobic compound with a main absorption wavelength of 460nm, and further modification is required to reach the phototherapy window. Past studies have shown that the structure of hypocrellin is easy to modify, and the modified derivatives can meet the requirements of strong light absorption in the phototherapy window (600-900nm) and water-solubility to meet the needs of clinical intravenous injection. Therefore, hypocrellin has broad application prospects as a photodynamic drug.

[0005] The research on the use of hypocrellin photosensitizer for photodynamic therapy of tumors is still in the research stage and there are few reports. Therefore, there is an urgent need to develop new hypocrellin photodynamic drugs for the treatment of tumor diseases. Summary of the invention

[0006] In order to improve the above technical problems, the present invention provides a hypocrellin derivative represented by formula (I), and its isomers, isotope-labeled substances or pharmaceutically acceptable salts:

[0007]

[0008] In formula (I), R1 is -H or -COCH3;

[0009] The general structural formula of R2 is -NH-(CH2) p —X—Y—(CH2CH2O) n —Z;

[0010] Wherein, X is a connecting group selected from -C 3-8 Cycloalkyl-, -(CH2CH2O) m -;

[0011] 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 group is connected to X, and the right side is connected to the polyethylene glycol unit;

[0012] Z is a terminal group selected from -H, -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;

[0013] p is selected from 0 or 1; m is selected from 2 or 3;

[0014] -(CH2CH2O) n - is a polyethylene glycol unit, n is selected from an integer between 1 and 100, for example, n is selected from an integer between 1 and 50;

[0015] In one embodiment, X is selected from cyclohexyl, cyclopentyl, cyclobutyl, cyclopropyl or -(CH2CH2O) m -, wherein m is selected from 2 or 3;

[0016] In one embodiment, 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)-, -C(=O)-CH2- or -C(=O)-CH2O-;

[0017] Z is selected from -H, -CH3, -C2H5, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H.

[0018] According to an embodiment of the present invention, when X is a cyclohexyl group, Y is selected from -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O- or -C(=O)NH-;

[0019] 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-;

[0020] 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-;

[0021] 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-;

[0022] When X is -(CH2CH2O) m -, m is selected from 2 or 3, and Y is selected from -C(=O)-, -C(=O)-CH2- or -C(=O)-CH2O-.

[0023] In one embodiment, R2 is selected from:

[0024] -NH-CH2C6H 10 -COO-(CH2CH2O) n -Z, -NH-CH2C6H 10 -COO-CH2-(CH2CH2O) n -Z,

[0025] -NH-CH2C6H 10 -CO-NH-(CH2CH2O) n -Z, -NH-CH2C6H 10 -O-CO-(CH2CH2O) n -Z,

[0026] -NH-C6H 10 -COO-(CH2CH2O) n -Z, -NH-C6H 10 -CO-NH-(CH2CH2O) n -Z,

[0027] -NH-C6H 10 -COO-CH2-(CH2CH2O) n -Z, -NH-C6H 10 -O-CO-(CH2CH2O) n -Z,

[0028] -NH-C6H 10 -O-CO-CH2-(CH2CH2O) n -Z, -NH-C6H 10 -O-CO-CH2O-(CH2CH2O) n -Z,

[0029] -NH-C5H8-COO-(CH2CH2O) n -Z, -NH-C5H8-CO-NH-(CH2CH2O) n -Z,

[0030] -NH-C5H8-COO-CH2-(CH2CH2O) n -Z、-NH-C5H8-O-CO-(CH2CH2O) n -Z、

[0031] -NH-C5H8-O-CO-CH2-(CH2CH2O) n -Z、-NH-C5H8-O-CO-CH2O-(CH2CH2O) n -Z、

[0032] -NH-C4H6-COO-(CH2CH2O) n -Z、-NH-C4H6-CO-NH-(CH2CH2O) n -Z、

[0033] -NH-C4H6-COO-CH2-(CH2CH2O) n -Z、-NH-C4H6-O-CO-(CH2CH2O) n -Z、

[0034] -NH-C4H6-O-CO-CH2-(CH2CH2O) n -Z、-NH-C4H6-O-CO-CH2O-(CH2CH2O) n -Z、

[0035] -NH-C3H4-COO-(CH2CH2O) n -Z、-NH-C3H4-CO-NH-(CH2CH2O) n -Z、

[0036] -NH-C3H4-COO-CH2-(CH2CH2O) n -Z、-NH-C3H4-O-CO-(CH2CH2O) n -Z、

[0037] -NH-C3H4-O-CO-CH2-(CH2CH2O) n -Z、-NH-C3H4-O-CO-CH2O-(CH2CH2O) n -Z、

[0038] -NH-(CH2CH2O) m -CO-(CH2CH2O) n -Z、-NH-(CH2CH2O) m -CO-CH2-(CH2CH2O) n -Z、

[0039] -NH-(CH2CH2O) m -CO-CH2O-(CH2CH2O) n -Z,

[0040] In the above groups, m is selected from 2 or 3; each n is the same or different and is independently selected from an integer of 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.

[0041] In one embodiment, when X is a cyclohexyl group, the group -(CH2) attached to the cyclohexyl group p - and group Y are located in the para, ortho or meta position; more preferably, the two groups are located in the para position;

[0042] In one embodiment, when X is a cyclopentyl group, the group -(CH2) attached to the cyclopentyl group p - and the group Y is located in the ortho or meta position;

[0043] In one embodiment, the terminal group Z is selected from -CH3, -H, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H;

[0044] In one embodiment, the R2 is, for example: Each n above is the same or different and is independently selected from an integer of 1 to 100;

[0045] Exemplarily, R2 is selected from:

[0046] In one embodiment, the R2 is, for example: Each n above is the same or different and is independently selected from an integer of 1 to 100;

[0047] Exemplarily, R2 is selected from

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

[0049] In one embodiment, R2 is, for example:

[0050] In one embodiment, R2 is, for example:

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

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

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

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

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

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

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

[0058] Each n above is the same or different and is independently selected from an integer of 1 to 100.

[0059] In one embodiment, the 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;

[0060] Each m above is selected from 2 or 3; each n is the same or different and independently selected from integers of 1 to 100;

[0061] Exemplarily, 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.

[0062] The general structural formula of the hypocrellin derivatives described in the above formula (I) also includes the enol tautomer shown in formula (I'). Therefore, although the present application only lists the structure of formula (I), the structure of formula (I') is also included in the protection scope of the present application. Those skilled in the art cannot exclude the structure of formula (I') from the protection scope of the present application because the present application does not list them one by one;

[0063]

[0064] The present invention also provides a method for preparing the derivative of formula (I), comprising the following steps:

[0065]

[0066] Wherein, R1, X, Y, Z, p, and n are as defined above; R3 is H, -COOH, -OH, or -NH2; R4 is H, -COOH, -OH, -CH2OH, -CH2COOH, -OCH2COOH, or -NH2;

[0067] 1) Compound (III) and NH2-(CH2) p -X-R3 reaction to obtain a compound of formula (II);

[0068] 2) Compound (II) and compound R4-(CH2CH2O) n -Z reacts to obtain a compound of formula (I), wherein R3 and R4 react to form a linking group Y.

[0069] According to the present invention, in the preparation method:

[0070] When R3 is -COOH, R4-(CH2CH2O) wherein R4 is -OH, -NH2, or -CH2OH is n -Z to react to obtain a compound of formula (I), wherein the connecting bond Y is -COO-, -CONH- or -COO-CH2-;

[0071] When R3 is -OH, R4-(CH2CH2O) of formula (II) and R4 is -COOH, -CH2COOH or -OCH2COOH n -Z to react to obtain a compound of formula (I), wherein the connecting bond Y is -O-CO-, -O-CO-CH2- or -O-CO-CH2O-;

[0072] When R3 is -NH, R4-(CH2CH2O) wherein R4 is -COOH, -CH2COOH or -OCH2COOH is n -Z to react to obtain a compound of formula (I), wherein Y is -NHCO-, -NHCO-CH2- or -NHCO-CH2O-;

[0073] When X is -(CH2CH2O) m - and R3 is -H, R4-(CH2CH2O) wherein R4 is -COOH, -CH2COOH or -OCH2COOH n -Z to react to obtain a compound of formula (I), wherein X is -(CH2CH2O) m -, Y is -CO-, -CO-CH2- or -CO-CH2O-.

[0074] In one embodiment, the reaction in step 1) and step 2) is carried out under the protection of an inert gas in the absence of light, and the inert gas is preferably argon or nitrogen.

[0075] In one embodiment, the compound of formula (III) in step 1) is hypocrellin B or deacetylhypocrellin.

[0076] In one embodiment, step 1) the compound of formula (III) and the compound NH2-(CH2) p The feeding 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, more preferably 1:60.

[0077] In one embodiment, the reaction temperature in step 1) is 50-120°C, more preferably 80°C.

[0078] In one embodiment, the reaction time of step 1) is 4 to 24 hours, more preferably 8 hours.

[0079] In one embodiment, the reaction in step 1) is carried out in a solvent, and the solvent is an organic solvent or a mixed solvent of an organic solvent and water. The mass fraction of water in the mixed solvent of organic solvent and water is 10wt% to 95wt%; 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; more preferably, the mixed solvent of organic solvent and water is a mixed solvent of N,N-dimethylformamide and water, wherein the volume ratio of N,N-dimethylformamide to water is 1:1.

[0080] In one embodiment, the reaction in step 1) can also be carried out under alkaline conditions, wherein the alkaline conditions are pH=10 to 14. The reagents used in the alkaline conditions are aqueous potassium hydroxide solution, aqueous sodium hydroxide solution, and aqueous potassium carbonate solution. More preferably, the reaction is carried out in a 1% aqueous sodium hydroxide solution.

[0081] In one embodiment, the reaction solution treatment process in step 1) is: removing the reaction organic solvent, dissolving the obtained blue-black solid residue with dichloromethane, washing three times with 5% dilute hydrochloric acid aqueous solution and once with water, drying the organic layer with anhydrous magnesium sulfate, filtering, and removing the solvent to obtain the compound of formula (II).

[0082] In one embodiment, step 2) the compound of formula (II) and the compound R4-(CH2CH2O) n -Z has a molar ratio of 1:1 to 10, exemplarily 1:1 to 5, for example 1:2;

[0083] In one embodiment, the reaction in step 2) can be carried out under the action of a condensing agent, and the condensing agent can be dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt) or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI);

[0084] In one embodiment, the reaction in step 2) can be carried out under the action of an organic base, and the organic base can be triethylamine, N,N-diisopropylethylamine or 4-dimethylaminopyridine (DMAP);

[0085] In one embodiment, the reaction in step 2) can be carried out in the presence of a solvent, and the solvent is an organic solvent, and the organic solvent can be tetrahydrofuran, dioxane, dichloromethane or chloroform;

[0086] In one embodiment, the reaction temperature in step 2) is 20 to 50° C., more preferably 25° C.;

[0087] In one embodiment, the reaction time of step 2) is 4 to 24 hours, more preferably 24 hours.

[0088] In one embodiment, the process of separating and purifying the compound of formula (I) in step 2) is as follows: removing the reaction organic solvent, dissolving the blue-black solid residue with dichloromethane, washing three times with a 5% dilute hydrochloric acid aqueous solution and once with water, drying the organic layer with anhydrous magnesium sulfate, filtering, and removing the solvent to obtain a crude product. The obtained crude product is further separated by silica gel plate chromatography, and the developing solvent is preferably a mixed solution of ethyl acetate, diethylamine and ethanol in a volume ratio of preferably 20:1:2 to obtain a derivative substituted with polyethylene glycol at the 2-position of hypocrellin, and the total yield of the two-step reaction is 3-15%, and the product is a blue-black solid.

[0089] In the present invention, the developing agent 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 agent used in silica gel plate chromatography is a mixed solution containing ethyl acetate, diethylamine and ethanol, and the volume ratio of ethyl acetate, diethylamine and ethanol in the mixed solution is 20:1:1 to 20:1:5.

[0090] The present invention also provides a pharmaceutical composition, which comprises the compound represented by formula (I) as described above, or at least one of its isomers, isotope-labeled substances or pharmaceutically acceptable salts.

[0091] According to an embodiment of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0092] According to an embodiment of the present invention, the pharmaceutical composition is used as 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.

[0093] The present invention also provides the use of the compound represented by formula (I), and at least one of its isomers, isotope-labeled substances or pharmaceutically acceptable salts in the preparation of drugs.

[0094] According to an embodiment of the present invention, the drug is an anti-tumor drug, such as a photodynamic therapy drug and / or a fluorescence-mediated drug and / or a photothermal therapy drug.

[0095] According to an embodiment of the present invention, the tumor is, for example, a digestive tract tumor, a head and neck facial tumor, a skin tumor, or a genital and urinary 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; the genital and urinary system tumor includes prostate cancer and bladder cancer.

[0096] The present invention also provides a method for treating tumors, comprising administering to an individual in need thereof a therapeutically effective amount of a compound represented by formula (I), at least one of its isomers, isotope-labeled substances or pharmaceutically acceptable salts.

[0097] The present invention also provides the use of the compound represented by formula (I), its isomers, isotope labels or pharmaceutically acceptable salts as fluorescence-mediated drugs in guiding the boundary resection of tumors.

[0098] The present invention also provides a method for guiding tumor resection, comprising administering to an individual in need thereof a therapeutically effective amount of a compound represented by formula (I), at least one of its isomers, isotope labels or pharmaceutically acceptable salts.

[0099] The maximum absorption wavelength of the hypocrellin derivatives of the present invention is about 580nm, and the molar extinction coefficient can reach 15000-20000nm. -1 cm -1 , has a strong light absorption capacity in the phototherapy window; the water-soluble polyethylene glycol-modified hypocrellin derivative disclosed in the present invention can efficiently produce singlet oxygen and other active oxygen species under photosensitization conditions, and can efficiently inactivate various tumor cells, such as esophageal cancer, gastric cancer, lung cancer, liver cancer, bile duct cancer, and colon cancer related to digestive tract tumors; head and neck cancer, brain cancer, tongue cancer, nasal cancer, oral cancer, and glioma related to head and neck facial tumors; basal cell carcinoma, squamous cell skin cancer, and melanoma related to skin tumors; and tumors related to the genitourinary system include prostate cancer and bladder cancer. Under the same conditions, the polyethylene glycol-substituted hypocrellin derivative photosensitizer involved in the present invention has a higher ability to photodynamically inactivate tumor cells than porphyrin commercial photosensitizers (HpD and Ce6). In addition, the present invention also discloses that such hypocrellin derivatives can not only produce active oxygen under laser irradiation, but also undergo significant photothermal conversion, and their photothermal conversion efficiency is between 25-35%, which can be used for photodynamic / photothermal synergistic treatment of tumors. Under the same conditions, the effect of photodynamic / photothermal synergistic therapy is better than that of photodynamic therapy alone. This application discloses for the first time that such hypocrellin derivatives have good photothermal conversion performance.

[0100] The water-soluble derivatives of the 2-position polyethylene glycol-substituted hypocrellin of the present invention have a solubility in physiological saline of more than 10 mg / mL. Such derivatives can be administered into the body through the tail vein as photosensitizing drugs, can specifically accumulate in tumor cells or tissues, can be used as photodynamic drugs to treat various tumors, or can be used as fluorescence-mediated drugs to assist in surgical resection of tumors.

[0101] Beneficial Effects

[0102] 1) The raw material of the hypocrellin in the present invention is extracted from natural products, which are easy to obtain, low in cost, can be prepared in large quantities, have little toxic side effects, and are easy to metabolize; the synthesis and separation methods are simple, without expensive reaction raw materials and complex separation methods; the product is easy to separate, purify, and has a clear structure, which overcomes the problems of porphyrin photosensitizers being difficult to separate, complex in composition, and difficult to determine in structure;

[0103] 2) The water-soluble derivative of the 2-position polyethylene glycol substituted hypocrellin described in the present invention has good water solubility and oil-water distribution ratio, which can fully meet the needs of clinical intravenous injection; it has strong red light absorption ability in the phototherapy window, and can efficiently produce reactive oxygen species under photosensitization conditions; it has a higher ability to photodynamically inactivate tumor cells than porphyrin photosensitizers; and the compound described in the present invention also has good photothermal conversion performance, and can generate heat to kill tumor cells under photothermal conditions. Therefore, the compound described in the present invention can have a photodynamic / photothermal synergistic treatment effect on tumor cells, and its effect is better than that of photodynamic therapy alone;

[0104] 3) The water-soluble derivatives of the 2-position polyethylene glycol-substituted bamboo red fungus described in the present invention can be used as photodynamic drugs in the treatment of digestive tract tumors, head and neck facial tumors, skin tumors, and genital and urinary 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; the genital and urinary system tumors include prostate cancer and bladder cancer. Such bamboo red fungus derivatives can effectively kill various tumor cells through intravenous injection as photosensitizers. Such photosensitizers at a concentration of 50nM can kill more than 90% of tumor cells, while having little effect on normal cells; such bamboo red fungus derivatives can be used as fluorescence-mediated drugs to guide tumor resection during surgery. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Figure 1 The general structural formula of the water-soluble derivative of hypocrellin substituted with polyethylene glycol at the 2-position of the present invention is shown.

[0106] Figure 2 The synthetic routes of the derivatives HB-1, HB-1-PEGn-CH3 (Example 2) and HB-1-NH-PEGn-CH3 (Example 12) are shown.

[0107] Figure 3 The synthetic routes of the derivatives HB-8 and HB-8-PEGn-CH3 (Example 28) are shown.

[0108] Figure 4 (a) Comparison of the absorption spectra of commercial photosensitizers PpIX and Ce6. Figure 4 (b) is a comparison diagram of the absorption spectra of HB, HB-1-PEG8-CH3 (Example 2), and HC-1-PEG8-CH3 (Example 7).

[0109] 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 superoxide radical scavenger.

[0110] Figure 6 (a) is the 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) is the photodegradation curve of the standard reference material Rose Bengal (RB); Figure 6 (d) is a comparison chart of the photodegradation of HB-1-PEG8-CH3, HC-1-PEG8-CH3 and RB.

[0111] Figure 7 (a) The derivatives of the present invention and commercial photosensitizers at 30 mW / cm 2 The photostability comparison chart of the laser illumination for 30 min, wherein 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; curve (iv) shows the photostability of the commercial photosensitizer HpD.

[0112] Figure 7 (b) The derivatives of the present invention and commercial photosensitizers at 30 mW / cm 2 The photostability comparison chart of the samples irradiated with laser for 30 min is shown in FIG. 1 , wherein curve (i) and curve (ii) respectively show the photostability of HB-8-PEG8-CH3 and HC-8-PEG8-CH3 (Example 28); curves (iii) and (iv) respectively show the photostability of commercial photosensitizers Ce6 and HpD.

[0113] Figure 8 (a) is a comparison chart of the pH stability of the derivatives of the present invention and commercial hematoporphyrin, wherein curve (i) shows the pH stability of the commercial photosensitizer HpD; curve (ii) and curve (iii) respectively show the pH stability of HB-1-PEG24-CO2CH3 (Example 5) and HC-1-PEG24-CO2CH3 (Example 10).

[0114] Figure 8(b) is a comparison chart of the pH stability of the derivatives of the present invention and the commercial photosensitizer HpD; wherein curve (i) shows the pH stability of HpD; curve (ii) and curve (iii) respectively show the pH stability of HB-8-PEG16-H (Example 29) and HC-8-PEG16-H (Example 29).

[0115] Fig. 9 (a) HB-1-PEG16-CH3 (Example 2) under different laser intensities (0.1-0.8 W cm -2 ) photothermal change curve 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 -2 Photothermal change curve under laser irradiation (temperature change of the solution within 10 minutes).

[0116] Fig.10 The confocal fluorescence imaging of HB-1-PEG16-CH3 in C6 cells prepared in Example 2 of the present invention; wherein Fig.10 (a) shows the dark field and bright field superimposed images, Fig.10 (b) shows a dark field image, Fig.10 (c) shows a bright field image.

[0117] Fig.11 The dark toxicity comparison graph (a) and phototoxicity comparison graph (b) of commercial HpD at different concentrations on esophageal cancer AKR cells, HB-1-PEG8-CH3 of Example 2 on esophageal cancer AKR cells, and HB-1-PEG16-CH3 on gastric cancer MFC cells are shown.

[0118] Fig.12 The dark toxicity comparison graph (a) and phototoxicity comparison graph (b) of commercial HpD at different concentrations on lung cancer cells A549, HC-1-PEG8-CH3 of Example 7 on lung cancer cells A549, and HC-1-PEG16-CH3 on liver cancer cells HCC are shown.

[0119] Fig.13 The dark toxicity comparison graph (a) and phototoxicity comparison graph (b) of different concentrations of commercial HpD on cholangiocarcinoma cells MCC, HB-8-PEG8-CH3 of Example 28 on cholangiocarcinoma cells MCC, and HB-8-PEG16-CH3 on colon cancer cells HCT116 are shown.

[0120] Fig.14 The phototoxicity graphs of commercial HpD and the compounds of the present invention at different concentrations are shown:

[0121] (a) is a comparison of the phototoxicity of commercial HPD to brain cancer cell G442, HC-8-PEG2-CH3 of Example 28 to brain cancer cell G442, and HC-8-PEG8-CH3 to head and neck cancer cell SCC2;

[0122] (b) is a comparison of the phototoxicity of commercial HpD on tongue cancer cell TSCCa, HC-8-PEG16-H of Example 29 on tongue cancer cell TSCCa, and HC-8-PEG32-H on nasal cancer cell KB;

[0123] (c) is a comparison of the phototoxicity of commercial HpD to oral cancer cells CAL27, the phototoxicity of HC-8-PEG2-SO3H in Example 31 to oral cancer cells CAL27, and the phototoxicity of HC-8-PEG8-SO3H to brain glioma cells C6.

[0124] Fig.15 The photodynamic effect of different concentrations of hypocrellin derivatives on tumor cells is shown in the figure:

[0125] (a) is a comparison of the 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;

[0126] (b) is a comparison 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;

[0127] (c) is a comparison of the 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;

[0128] (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;

[0129] (e) is a comparison of the phototoxicity of HB-7-PEG4, HC-7-PEG16 of Example 27 and HB-8-PEG32-CO2CH3 of Example 30 to bladder cancer MBT-2 cells;

[0130] (f) is a comparison chart 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 on basal cell carcinoma BCC cells.

[0131] Fig.16 The photodynamic effect diagram 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.

[0132] Fig.17 The in vivo fluorescence imaging diagrams of the derivatives of the present invention in mice with different tumor models 4 hours after 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.

[0133] Definition and explanation of terms

[0134] Unless otherwise defined, all technical terms herein have the same meaning as those generally understood by a person skilled in the art to which the subject matter of the claims belongs. When the numerical ranges recorded in the specification and claims of this application are defined as "integers", they should be understood to record the two endpoints of the range and every integer in the range. For example, "an integer from 1 to 6" should be understood to record every integer of 1, 2, 3, 4, 5 and 6, and so on.

[0135] The term "C 1-6 "Alkyl" is understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 6 carbon atoms. For example, "C 1-6 The term "alkyl" refers to straight-chain and branched alkyl groups 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.

[0136] The term "C 1-6 "Alkoxy" is understood to mean -OC 1-6 Alkyl, where C 1-6 Alkyl has the above definition.

[0137] The term "C 3-8 "Cycloalkyl" is understood to mean a saturated monovalent cycloalkyl radical, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl.

[0138] It should also be noted that the hypocrellin derivatives to be protected in the present invention all contain two enol tautomers, and the chemical structures of the two isomers such as formula (I) and formula (I') are of course within the scope of protection. For the sake of simplicity, only one of the enol tautomers is listed in all embodiments of the present invention, and the other enol tautomer and its corresponding general structural formula are described in detail in the specification, and its structure is of course within the scope of protection. In addition, the general structural formula of the hypocrellin derivatives involved in the present invention includes a polyethylene glycol unit (PEGn), and its unit number n is any integer between 1 and 100, and its corresponding chemical structure is of course within the scope of protection. For the sake of simplicity, only some integers are listed in all embodiments of the present invention, and the general structural formula corresponding to the remaining parts is described in detail in the specification, and its structure is of course within the scope of protection. Any range recorded in the present invention includes the end value and any numerical value between the end values ​​and any sub-range composed of the end value or any numerical value between the end values. DETAILED DESCRIPTION

[0139] In order to more clearly illustrate the present invention, the present invention is further described below in conjunction with preferred embodiments and accompanying drawings. Those skilled in the art should understand that the content described specifically below is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention. In the present invention, the experimental methods described are all conventional methods unless otherwise specified. The raw materials used can be obtained from public commercial channels unless otherwise specified; the percentages are all mass percentages unless otherwise specified; the M is mol / L unless otherwise specified. The bamboo red fungus derivatives involved in the present invention are shown in Examples 2 to 36, and the characterization results are shown in Table 1.

[0140] Example 1

[0141] The raw materials used in the present invention are hypocrellin A HA, hypocrellin B HB, and deacetyl hypocrellin HC, and their molecular structures are shown below:

[0142]

[0143] Example 2

[0144] Tranexamic acid-polyethylene glycol substituted hypocrellin B derivatives (R1=-COCH3, R2=-NHCH2-C6H 10 -COO-PEGn-CH3) (PEGn is polyethylene glycol, n = 2, 8, 12, 16, 20, 24) preparation: Hypocrellin B HB (1 mmol), tranexamic acid H2N-CH2C6H 10COOH (50mmol) and NaOH (200mmol) were dissolved in 100mL DMF and water mixed solution (volume ratio of 1:1), heated to 120℃ under nitrogen protection and reacted in the dark for 6h. After the reaction, dilute hydrochloric acid was added to adjust the pH to weak acidity and a large amount of black solid was precipitated, which was filtered to obtain the crude product HB-1. The above crude product was added with dicyclohexylcarbodiimide DCC (1mmol) and 4-dimethylaminopyridine DMAP (1mmol), dissolved in 50mL anhydrous dichloromethane, and reacted with polyethylene glycol methyl esters of different chain lengths (HO-PEGn-CH3, 1mmol), respectively, and stirred at room temperature in the dark for 12h. After the reaction, the reaction solution was washed with 100mL dilute hydrochloric acid aqueous solution, washed three times with distilled water, dried, filtered, and the crude product was separated by thin layer chromatography, and the developing solvent was ethyl acetate and ethanol = 5:1 to obtain a 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 the synthesis route thereof is as follows:

[0145] Example 3

[0146] Tranexamic acid-HB derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 Preparation of HB-1-PEGn-H (n=4, 12, 18): The preparation method is basically the same as that in Example 2, except that polyethylene glycol (HO-PEGn-H) of different chain lengths is used to replace polyethylene glycol methyl ether to obtain a blue-black solid product HB-1-PEGn-H (n=4, 12, 18).

[0147] Example 4

[0148] Tranexamic acid-HB derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 -COO-PEGn-CH2COOH): The preparation method is the same as Example 2, except that HO-PEGn-CH2COOH of different chain lengths is used to replace polyethylene glycol methyl ether to obtain a blue-black solid product HB-1-PEGn-CO2H (n=2, 8, 16).

[0149] Example 5

[0150] Tranexamic acid-HB derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10-COO-PEGn-CH2CH2COOCH3): The preparation method is basically the same as Example 2, replacing polyethylene glycol methyl ether with HO-PEGn-CH2CH2COOCH3 to obtain a blue-black solid product HB-1-PEGn-CO2CH3 (n=8, 16, 24).

[0151] Example 6

[0152] Tranexamic acid-HB derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 -COO-PEGn-CH2CH2SO3H): The preparation method is basically the same as Example 2, except that ethylene glycol methyl ether is replaced by HO-PEGn-CH2CH2SO3H of different chain lengths to obtain a blue-black product HB-1-PEGn-SO3H (n=2, 8, 16).

[0153] The characterization results of the amino-substituted products in Examples 3 to 6 are shown in Table 1, and their molecular structures are shown below:

[0154]

[0155] Example 7

[0156] Tranexamic acid-HC derivatives substituted with polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 Preparation of HC-1-PEGn-CH3): The preparation method is basically the same as Example 2, except that HC is used to replace HB and HO-PEGn-CH3 is used to replace polyethylene glycol methyl ether to obtain a 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 the synthesis route is as follows:

[0157]

[0158] Example 8

[0159] Tranexamic acid-HC derivatives substituted with polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10Preparation of HC-1-COO-PEGn-H): HC-1 synthesized in Example 7 was used as a raw material, and reacted with polyethylene glycol (HO-PEGn-H) of different chain lengths, respectively, and stirred at room temperature in the dark for 12 hours. 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 solution 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, and the developing solvent was a mixture of ethyl acetate and ethanol = 5:1, to obtain a blue-black solid product HC-1-PEGn-H (n = 2, 12, 16).

[0160] Example 9

[0161] Tranexamic acid-HC derivatives substituted with polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 -COO-PEGn-CH2COOH): HC-1 synthesized in Example 7 was used as a raw material, the polyethylene glycol used was HO-PEGn-CH2COOH, and the synthesis method was similar to the synthesis method of HC-1-PEGn-CH3 in Example 7 to obtain a blue-black solid product HC-1-PEGn-CO2H (n=2, 8, 24).

[0162] Example 10

[0163] Tranexamic acid-HC derivatives substituted with polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 -COO-PEGn-CH2CH2COOCH3): HC-1 synthesized in Example 7 was used as a raw material, the polyethylene glycol used was HO-PEGn-CH2CH2COOCH3, and the synthesis method was similar to the synthesis method of HC-1-PEGn-CH3 in Example 7 to obtain a blue-black solid product HC-1-PEGn-CO2CH3 (n=2, 16, 24).

[0164] Embodiment 11

[0165] Tranexamic acid-HC derivatives substituted with polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 -COO-PEGn-CH2CH2SO3H): HC-1 synthesized in Example 7 was used as a raw material, the polyethylene glycol used was HO-PEGn-CH2CH2SO3H, and the synthesis method was similar to the synthesis method of HC-1-PEGn-CH3 in Example 7 to obtain a blue-black solid product HC-1-PEGn-SO3H (n=2, 8, 24).

[0166] The characterization results of the amino-substituted products in Examples 8 to 11 above are shown in Table 1, and their molecular structures are shown below:

[0167]

[0168] Example 12

[0169] Tranexamic acid-HB derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 Preparation of amino polyethylene glycol: The amino polyethylene glycol used is H2N-PEGn-CH3, and the synthesis method is similar to the synthesis method of HB-1-PEGn-CH3 in Example 2, and a blue-black solid product HB-1-NH-PEGn-CH3 (n=4, 8, 16, 24) is obtained. The characterization results of the above amino-substituted products are shown in Table 1, and the synthesis route is as follows:

[0170]

[0171] Example 13

[0172] Tranexamic acid-HB derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 Preparation of HB-1-NH-PEGn-H): HB-1 synthesized in Example 2 was used as a raw material, and reacted with polyethylene glycols (H2N-PEGn-H, 0.5 mmol) of different chain lengths, respectively, and stirred at room temperature in the dark for 12 hours. 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 solution 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, and the developing solvent was a mixture of ethyl acetate and ethanol = 5:1, to obtain a blue-black solid product HB-1-NH-PEGn-H (n = 2, 16, 24).

[0173] Embodiment 14

[0174] Tranexamic acid-HB derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 -CO-NH-PEGn-CH2COOH): HB-1 synthesized in Example 2 was used as the raw material, the polyethylene glycol used was H2N-PEGn-CH2COOH, and the synthesis method was similar to the synthesis method of HB-1-PEGn-H in Example 3 to obtain a blue-black solid product HB-1-NH-PEGn-CO2H (n=8, 16, 24).

[0175] Embodiment 15

[0176] Tranexamic acid-HB derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H10 -CO-NH-PEGn-CH2CH2COOCH3): HB-1 synthesized in Example 2 was used as raw material, the polyethylene glycol used was H2N-PEGn-CH2CH2COOCH3, and the synthesis method was similar to the synthesis method of HB-1-PEGn-H in Example 3 to obtain a blue-black solid product HB-1-NH-PEGn-CO2CH3 (n=2, 8, 16).

[0177] Example 16

[0178] Tranexamic acid-HB derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C6H 10 -CO-NH-PEGn-CH2CH2SO3H): HB-1 synthesized in Example 2 was used as the raw material, the polyethylene glycol used was H2N-PEGn-CH2CH2SO3H, and the synthesis method was similar to the synthesis method of HB-1-PEGn-H in Example 3 to obtain a blue-black solid product HB-1-NH-PEGn-SO3H (n=2, 8, 16).

[0179] The characterization results of the amino-substituted products in Examples 13 to 16 are shown in Table 1, and their molecular structures are shown below:

[0180]

[0181] Embodiment 17

[0182] Tranexamic acid-HC derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 Preparation of HC-1-NH-PEGn-CH3): HC is used as the hypocrellin, NH2-PEGn-CH3 is used as the polyethylene glycol, and the synthesis method is similar to the synthesis method of HB-1-PEGn-CH3 in Example 2 to obtain 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 the synthesis route is as follows:

[0183]

[0184] Embodiment 18

[0185] Tranexamic acid-HC derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10Preparation of HC-1-NH-CO-NH-PEGn-H): HC-1 synthesized in Example 7 was used as a raw material, and reacted with polyethylene glycols of different chain lengths (H2N-PEGn-H, 0.5 mmol) respectively, and stirred at room temperature in the dark for 12 hours. 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 solution and three times with distilled water. The organic layer was dried and filtered, and the crude product was separated by thin layer chromatography with a developing solvent of ethyl acetate and ethanol = 5:1 to obtain a blue-black solid product HC-1-NH-PEGn-H (n = 2, 8, 24).

[0186] Embodiment 19

[0187] Tranexamic acid-HC derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 -CO-NH-PEGn-CH2COOH): HC-1 synthesized in Example 7 was used as the raw material, and the polyethylene glycol used was H2N-PEGn-CH2COOH. The synthesis method was similar to that of HC-1-PEGn-CH3 to obtain a blue-black solid product HC-1-NH-PEGn-COOH (n=8, 16, 24).

[0188] Embodiment 20

[0189] Tranexamic acid-HC derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 -CO-NH-PEGn-CH2CH2COOCH3): Using HC-1 synthesized in Example 7 as raw material and polyethylene glycol H2N-PEGn-CH2CH2COOCH3, a blue-black solid product HC-1-NH-PEGn-CO2CH3 (n=8, 16, 24) was obtained by a synthesis method similar to that of HC-1-PEGn-CH3.

[0190] Embodiment 21

[0191] Tranexamic acid-HC derivatives substituted with amino polyethylene glycol of different chain lengths (R1=-H, R2=-NHCH2-C6H 10 -CO-NH-PEGn-CH2CH2SO3H): HC-1 synthesized in Example 7 was used as the raw material, and the polyethylene glycol used was H2N-PEGn-CH2CH2SO3H. The synthesis method of HC-1-PEGn-CH3 was similar to that of HC-1-PEGn-CH3 to obtain a blue-black solid product HC-1-NH-PEGn-SO3H (n=2, 8, 16).

[0192] 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:

[0193]

[0194] Embodiment 22

[0195] O-aminocyclic acid-HB derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NH-C6H 10 Preparation of HB-2-o-PEGn (n=4, 8, 16): The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to the synthesis method of HB-1-PEGn in Example 2 to obtain a 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 the synthesis route is as follows:

[0196]

[0197] HB derivatives substituted by meta-aminocyclic acid and polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NH-C6H 10 Preparation of HB-2-m-PEGn (n=4, 8, 16): The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to the synthesis method of HB-1-PEGn in Example 2 to obtain 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 the synthesis route is as follows:

[0198]

[0199] HB derivatives substituted with aminocyclic acid and polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NH-C6H 10 Preparation of HB-1-PEGn-COO-PEGn-CH3): The polyethylene glycol used was HO-PEGn-CH3, and the synthesis method was similar to the synthesis method of HB-1-PEGn in Example 2 to obtain a 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 the synthesis route is as follows:

[0200]

[0201] Embodiment 23

[0202] Preparation of aminocyclobutyric acid-HB or HC derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3 or -H, R2=-NH-C4H6-COO-PEGn-CH3): The polyethylene glycol used is HO-PEGn-CH3, and the synthesis method is similar to the synthesis method of HB-1-PEGn in Example 2, and the hypocrellin derivative HB-3-PEGn (n=4, 8, 16) and the deacetylated hypocrellin derivative HC-3-PEGn (n=4, 8, 16) are obtained. The characterization results of the above-mentioned aminated products are shown in Table 1, and the synthesis route is as follows:

[0203] Embodiment 24

[0204] Preparation of aminocyclopropane-HB derivatives substituted with polyethylene glycol of different chain lengths (R1=-COCH3, R2=-NHCH2-C3H4-O-CO-PEGn-CH3): The polyethylene glycol used was HOOC-PEGn-CH3, and the synthesis method was similar to the synthesis method of HB-1-PEGn in Example 2 to obtain a 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 the synthesis route is as follows:

[0205]

[0206] Embodiment 25

[0207] Preparation of HB or HC derivatives (R1=-COCH3, R2=-NH-C5H8-COO-PEGn-CH3) substituted by m-aminocyclopentanoic acid and polyethylene glycol of different chain lengths: The polyethylene glycol used is HO-PEGn-CH3, and the synthesis method is similar to the synthesis method of HB-1-PEGn in Example 2, and blue-black solid products HB-5-PEGn (n=2, 10, 20) and HC-5-PEGn (n=2, 10, 20) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0208]

[0209] Embodiment 26

[0210] p-Aminocyclohexanol-HB or HC derivatives substituted with polyethylene glycol of different chain lengths (R1 = -COCH3 or -H, R2 = -NH-C6H 10-OCO-PEGn-CH3) preparation: The polyethylene glycol used is HOOC-PEGn-CH3, and the synthesis method is similar to the synthesis method of HB-1-PEGn in Example 2, and the hypocrellin B derivative HB-6-PEGn (n=1, 8, 24) and the deacetyl hypocrellin derivative HC-6-PEGn (n=1, 8, 24) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0211] Embodiment 27

[0212] p-Aminocyclohexanol-HB or HC derivatives substituted with polyethylene glycol of different chain lengths (R1 = -COCH3 or -H, R2 = -NH-C6H 10 Preparation of HB-6 or HC-6 prepared in Example 26, and HOOC-CH2O-PEGn-CH3 as the raw material, to obtain the hypocrellin B derivative HB-7-PEGn (n=4, 8, 16) and the deacetyl hypocrellin derivative HC-7-PEGn (n=4, 8, 16). The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0213] Embodiment 28

[0214] Preparation of ethanolamine-different chain length polyethylene glycol substituted HB derivatives (R1=-COCH3, R2=-NH-(CH2CH2-O)2-CO-PEGn-CH3)(n=2,8,16,32): HB (1mmol), glycoside diol NH2-CH2CH2-O-CH2CH2-OH (50mmol) and NaOH (100mmol) were dissolved in a mixed solution of 100mL DMF and water (volume ratio of 1:1), and after thorough mixing, heated to 120℃ under nitrogen protection, stirred in the dark for 6h. After the reaction was completed, dilute hydrochloric acid was added to adjust the pH of the reaction solution to weak acidity 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 with DCC (1 mmol) and DMAP (1 mmol), dissolved in 50 mL of anhydrous dichloromethane, and reacted with polyethylene glycol methyl esters (HO-PEGn-OCH3, 1 mmol) of different chain lengths, respectively, and stirred at room temperature in the dark for 12 hours. After the reaction was completed, the reaction solution was washed once with 100 mL of dilute hydrochloric acid solution and three times with distilled water, the organic layer was dried with anhydrous magnesium sulfate, filtered, and the crude product was separated by thin layer chromatography, the developing solvent was ethyl acetate and ethanol = 5:1, and a blue-black solid product HB-8-PEGn-CH3 (n = 2, 8, 16, 32) was obtained.

[0215] Preparation of HC derivatives (R1=-H, R2=-NH-(CH2CH2-O)2-CO-PEGn-CH3) (n=2,8,16,32) substituted by ethanolamine-polyethylene glycol of different chain lengths: HC was used to replace HB, and the above experiment was repeated to obtain a 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 the synthesis route is as follows:

[0216]

[0217] Embodiment 29

[0218] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-CO-PEGn-H) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: HB-8 or HC-8 synthesized in Example 28 is used as raw material, and the polyethylene glycol used is HOOC-PEGn-H. Similar to the synthesis method of HB-8-PEGn-CH3, the hypocrellin B derivative HB-8-PEGn-H (n=2, 16, 32) and the deacetyl hypocrellin derivative HC-8-PEGn-H (n=8, 16, 32) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0219]

[0220] Embodiment 30

[0221] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-CO-PEGn-CH2CH2CO2CH3) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: Using HB-8 or HC-8 synthesized in Example 28 as raw materials, the polyethylene glycol used is HOOC-PEGn-CH2CH2CO2CH3, and similar to the synthesis method of HB-8-PEGn-CH3, the hypocrellin B derivative HB-8-PEGn-CO2CH3 (n=2, 8, 32) and the deacetyl hypocrellin derivative HC-8-PEGn-CO2CH3 (n=8, 16, 32) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0222]

[0223] Embodiment 31

[0224] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-CO-PEGn-CH2CH2SO3H) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: HB-8 or HC-8 synthesized in Example 28 is used as raw material, and the polyethylene glycol used is HOOC-PEGn-CH2CH2SO3H. The synthesis method is similar to the synthesis method of HB-8-PEGn-CH3 in Example 28, and the hypocrellin B derivative HB-8-PEGn-SO3H (n=2, 8, 16) and the deacetylated hypocrellin derivative HC-8-PEGn-SO3H (n=2, 8, 16) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0225]

[0226] Embodiment 32

[0227] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-COCH2-PEGn-CH3) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: HB-8 or HC-8 synthesized in Example 28 is used as raw material, and the polyethylene glycol used is HOOC-CH2-PEGn-OCH3. Similar to the synthesis method of HB-8-PEGn-CH3, the hypocrellin B derivative HB-9-PEGn-CH3 (n=2, 8, 16) and the deacetylated hypocrellin derivative HC-9-PEGn-CH3 (n=2, 8, 16) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0228]

[0229] Embodiment 33

[0230] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-COCH2-PEGn-H) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: HB-8 or HC-8 synthesized in Example 28 is used as raw material, and the polyethylene glycol used is HOOC-CH2-PEGn-H. Similar to the synthesis method of HB-8-PEGn-CH3, the hypocrellin derivative HB-9-PEGn-H (n=2,8,16) or the deacetylated hypocrellin derivative HC-9-PEGn-H (n=2,8,16) is obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0231]

[0232] Embodiment 34

[0233] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-COCH2-O-PEGn-CH3) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: HB-8 or HC-8 synthesized in Example 28 is used as raw material, and the polyethylene glycol used is HOOC-CH2-O-PEGn-CH3. Similar to the synthesis method of HB-8-PEGn-CH3, the hypocrellin derivative HB-10-PEGn-CH3 (n=2, 8, 16) and the deacetylated hypocrellin derivative HC-10-PEGn-CH3 (n=2, 8, 16) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0234]

[0235] Embodiment 35

[0236] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)2-COCH2-O-PEGn-H) substituted with glycoside diol amine and polyethylene glycol of different chain lengths: Using HB-8 or HC-8 synthesized in Example 28 as raw materials, similar to the synthesis method of HB-8-PEGn-CH3, the hypocrellin derivative HB-10-PEGn-H (n=2,8,16) or the deacetylated hypocrellin derivative HC-10-PEGn-H (n=2,8,16) was obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0237]

[0238] Embodiment 36

[0239] Preparation of HB or HC derivatives (R1=-COCH3 or -H, R2=-NH-(CH2CH2-O)3-CO-PEGn-CH3) substituted with glycoside triol amine and polyethylene glycol of different chain lengths: the amino substituent used is H2N-(CH2CH2-O)3-H, the polyethylene glycol used is HOOC-PEGn-CH3, and the synthesis method is similar to the synthesis method of HB-8-PEGn-CH3 in Example 28, and the hypocrellin B derivative HB-11-PEGn (n=8, 16, 24) and the deacetylated hypocrellin derivative HC-11-PEGn (n=8, 16, 24) are obtained. The characterization results of the above derivatives are shown in Table 1, and the synthesis route is as follows:

[0240]

[0241] Embodiment 37

[0242] 1) Absorption spectrum

[0243] Figure 4 The absorption spectra of the compounds related to the present invention are shown. Figure 4 In (a), the commercial porphyrin photosensitizer PpIX has multi-band absorption, all of which are narrow absorption bands. The maximum absorption wavelengths that can be used for phototherapy are 570nm and 630nm, and their molar extinction coefficients are all lower than 8000M. -1 cm -1 The maximum absorption wavelength of commercial porphine photosensitizer Ce6 is 650nm, and the molar extinction coefficient is about 15000M -1 cm -1 , which also has narrow absorption in the phototherapy window, so commercial PpIX and Ce6 have limited light absorption capacity in the phototherapy window. However, the absorption spectrum properties of the hypocrellin and its polyethylene glycol modified product disclosed in the present invention are completely different. Figure 4 As shown in (b), the maximum absorption peak of HB is around 470nm. The derivative HB-1-PEG8-CH3 prepared in Example 2 has a wide strong absorption in the phototherapy window. It has a wide absorption band between 500-750nm. It is a strong continuous absorption band in the phototherapy window. The maximum absorption peak is around 580nm, which is about 110nm red-shifted than the maximum absorption peak of HB. The molar extinction coefficient is about 20000M. -1 cm -1 Around, showing strong red light absorption ability. Similarly, the absorption spectrum of the derivative HC-1-PEG8-CH3 prepared in Example 7 is also located in the ideal phototherapy window, and it has a wide absorption band between 500-750nm, and the maximum absorption peak is around 580nm, showing strong red light absorption ability. The absorption spectrum of other bamboo red mycelium derivatives of the present invention is also similar to HB-1-PEG8-CH3, with a wide absorption band between 500-750nm, and the maximum absorption peak is around 580nm. Therefore, the absorption wavelength of the 2-position polyethylene glycol-substituted water-soluble derivative of bamboo red mycelium described in the present invention and its light absorption ability in the phototherapy window are far superior to the commercial photosensitizers PpIX and Ce6, showing more prominent red light absorption ability. The maximum absorption wavelength of some compounds described in the present invention is shown in Table 1.

[0244] 2) Reactive oxygen species

[0245] The active oxygen species of the derivatives of the present invention were determined by electrospinning resonance (ESR). Figure 5The ESR action diagram of the active oxygen species of the derivative HB-1-PEG16-CH3 prepared in Example 2 is shown. The experiment shows that HB-1-PEG16-CH3 can efficiently generate reactive oxygen species (ROS). The singlet oxygen and superoxide radical scavengers are used to measure that this type of derivative can efficiently generate photosensitive active species, mainly singlet oxygen ( Figure 5 a), can also produce a small amount of superoxide free radicals ( Figure 5 b) Both active oxygen species are beneficial to photodynamic therapy. The derivatives disclosed in other embodiments of the present invention also have the ability to efficiently generate singlet oxygen and assist in generating a small amount of superoxide free radicals.

[0246] 3) Singlet oxygen yield

[0247] Singlet oxygen quantum yield test: 9,10-anthraquinone malonate sodium (ADPA) was used as a singlet oxygen scavenger and methylene blue (MB) was used as a standard photosensitizer (singlet oxygen yield was 0.52). ADPA solution was added to the derivative of the present invention or MB solution and the singlet oxygen quantum yield was measured at 635 nm (0.1 W cm -2 ) laser was used as the light source, the absorption intensity of ADPA at 378 nm was plotted against the irradiation time, and the singlet oxygen yield of the derivative was calculated. Figure 6 (a) and 6(b) show the photodegradation curves of the derivatives HB-1-PEG8-CH3 (Example 2) and HC-1-PEG8-CH3 (Example 7), respectively. It can be seen that the photosensitizer obviously produces singlet oxygen under light conditions, thereby significantly degrading ADPA. By comparing and calculating the singlet oxygen efficiency curve of the reference material Rose Bengal RB ( Figure 6 (c) and Figure 6 (d)), the singlet oxygen efficiency of HB-1-PEG8-CH3 and HC-1-PEG8-CH3 are 0.32 and 0.36 respectively. The efficiency of producing singlet oxygen of the hypocrellin derivatives disclosed in other embodiments of the present invention is between 0.2-0.4, and they also have the ability to efficiently produce active oxygen. The singlet oxygen yield of the compounds of the present invention is shown in Table 1.

[0248] 4) Water solubility

[0249] The hypocrellin derivatives disclosed in the present invention all contain hydrophilic groups of polyethylene glycol, so that the photosensitizer molecules have strong water solubility under physiological conditions. Experiments show that each milliliter of physiological saline or glucose injection can dissolve more than 10 mg of the photosensitizer of the present invention, showing excellent water solubility, so that the derivatives of the present invention can be well transported in blood vessels during intravenous injection without causing blood vessel blockage. HB-1-PEG8-CH3 (Example 2) contains 8 ethylene glycol units and a methyl terminal group, and can dissolve more than 10 mg per milliliter of normal saline; HB-1-PEG18-H (Example 3) contains 18 ethylene glycol units and a hydrogen terminal group, and can dissolve more than 15 mg per milliliter of normal saline; HC-1-PEG16-H (Example 8) contains 16 ethylene glycol units and can dissolve more than 15 mg per milliliter of normal saline; HB-8-PEG16-CH3 (Example 28) contains 16 ethylene glycol units and a methyl terminal group, and can dissolve more than 20 mg per milliliter of normal saline; HC-8-PEG16-CO2CH3 (Example 30) contains 16 ethylene glycol units and an ester terminal group, and can dissolve more than 15 mg per milliliter of normal saline; these compounds all show excellent water solubility. The derivatives disclosed in other embodiments of the present invention also have good water solubility and biocompatibility. Each milliliter of physiological saline can dissolve more than 3 to 20 mg of the derivatives of the present invention.

[0250] 5) Photostability

[0251] Comparison of the photostability of the derivatives prepared by the present invention and commercial photosensitizers Figure 7 As shown. Figure 7 As can be seen in (a), the 635nm laser at 30mW / cm 2 After irradiation at 500 nm light intensity for 30 min, the absorption spectrum of HB-1-PEG16-CH3 (Example 2) did not decrease significantly, and the absorption intensity of its maximum wavelength decreased by less than 5% (curve i). Under the same conditions, the absorption intensity of the maximum wavelength of HC-1-PEG16-CH3 (Example 7) also decreased by less than 10%, and it also had good photostability (curve ii). Under the same conditions, using a 635 nm laser at 30 mW / cm 2 After 30 minutes of illumination at a low light intensity, the maximum absorption of the commercial porphin photosensitizer Ce6 decreased by 30% (curve iii); while the absorption spectrum of the commercial hematoporphyrin photosensitizer HpD decreased even more, reaching about 50% (curve iv). Figure 7It can be seen from (b) that under the same conditions, the absorption intensity at the maximum wavelength of HB-8-PEG8-CH3 (Example 28) decreases by less than 10%, and has good photostability (curve i); the absorption intensity at the maximum wavelength of HC-8-PEG8-CH3 (Example 28) also decreases by less than 10%, and also has good photostability (curve ii). The derivatives disclosed in other embodiments of the present invention also have good photostability, and the absorption intensity of the maximum wavelength under the same conditions decreases by less than 10%. Therefore, the hypocrellin derivatives prepared by the present invention have better photostability than commercial photosensitizers.

[0252] 6) pH stability

[0253] The pH stability of the hypocrellin derivatives of the present invention is as follows Figure 8 As shown. The absorption spectrum of this type of derivatives has no obvious change in the pH range of 6.2 to 8.0, indicating that this type of derivatives has 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) has no obvious change in the pH range of 6.2 to 8.0; curve (iii) shows that the absorption spectrum of HC-1-PEG24-CO2CH3 (Example 10) has no obvious change in the pH range of 6.2 to 8.0; it has good pH stability in this range, because the two phenolic hydroxyl groups of bamboo red fungus are not easy to deprotonate under this acid-base condition; and the commercial 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, thus showing the instability of the HpD photosensitizer (curve i). Similarly, Figure 8 As shown in (b), curve (ii) shows that the absorption spectrum of HB-8-PEG16-H (Example 29) has no obvious change in the pH range of 6.2 to 8.0; curve (iii) shows the maximum absorption curve of HC-8-PEG16-H (Example 29) with pH change, which also has good pH stability in this range, also because the two phenolic hydroxyl groups of hypocrellin are not easy to deprotonate under this acid-base condition. The derivatives disclosed in other embodiments of the present invention also have good pH stability under physiological conditions, and the change of solution pH in the range of 6.2 to 8.0 has little effect on the absorption spectrum of photosensitizers, and has better pH stability than commercial HpD.

[0254] 7) Photothermal conversion efficiency

[0255] The derivatives of the present invention were exposed to lasers of different intensities (0.1-0.8 W cm) at a wavelength of 635 nm. -2) to detect its temperature change under irradiation. Photothermal conversion efficiency test: Add solutions of derivatives with different concentrations (0-50μM, 1.0mL) into a quartz cuvette, and measure the photothermal performance under laser irradiation of a certain wavelength and intensity. Use a thermocouple to record the temperature change through a digital thermometer, and use a Fotric220 infrared thermal imager to record the real-time thermal imaging of the sample. The solution of bamboo red fungus was irradiated with a 635nm laser (0.8Wcm -2 ) irradiated for 15 min and cooled naturally, and the photothermal conversion efficiency was measured. The photothermal conversion efficiency η was calculated by the formula:

[0256]

[0257] Where h is the heat transfer coefficient, A is the surface area of ​​the quartz cell, and T max is the maximum temperature at which the derivative solution of the present invention is heated, T surr is room temperature, Q dis is the heat loss caused by the absorption of light by the quartz sample cell and the solvent, I is the laser power, A 635 It is the absorbance of the hypocrellin derivative of the present invention at 635 nm.

[0258] The hypocrellin derivative prepared by the invention exhibits good photothermal conversion efficiency under 635nm laser irradiation.

[0259] The derivative HB-1-PEG16-CH3 (Example 2) was exposed to lasers of different intensities (0.1-0.8 W cm -2 ) under irradiation, the light-heat change curve is as follows Fig. 9 (a). When the laser intensity is 0.1 W cm -2 When the HB-1-PEG16-CH3 solution (concentration of 10 μM) was irradiated for 10 min, its temperature increased by only 3.0 °C, and the photothermal effect was negligible. -2 (all other experimental conditions were the same), the temperature of the solution increased by 18.5°C; when the light intensity continued to increase to 0.8 W cm -2 When the temperature of the solution was increased by 28.8℃, a significant photothermal effect was produced. This shows that the HB-1-PEG16-CH3 solution under laser irradiation can simultaneously produce singlet oxygen and photothermal effect, and can be used for photodynamic / photothermal synergistic therapy.

[0260] The derivatives prepared by the present invention have a high sensitivity at 635nm and 0.8W cm -2 The photothermal change curve under laser irradiation is as follows Fig. 9 (b) As shown in the figure, pure water was used as the blank control group, and the temperature of pure water only rose by 4.5°C; the derivative HB-1-PEG16-CH3 prepared in Example 2 -2Under laser irradiation, the temperature of the solution increased by 28.8°C, producing an obvious photothermal effect with a photothermal conversion efficiency of 35%. -2 Under laser irradiation, the temperature of the solution increased by 25.8°C, producing an obvious photothermal effect with a photothermal conversion efficiency of 33%. -2 Under laser irradiation, the temperature of the solution increased by 23.5°C, producing a significant photothermal effect, and its photothermal conversion efficiency was 31%; under the same conditions, the temperature of the solution of HC-1-PEG12-H prepared in Example 8 increased by 22.4°C, producing a significant photothermal effect, and its photothermal conversion efficiency was 30.8%. In addition to the above derivatives, other derivatives of the present invention also have good photothermal conversion performance, and their photothermal conversion efficiency is between 25% and 35%. See Table 1 for specific photothermal conversion efficiency.

[0261] Table 1: Characterization data of the hypocrellin derivatives prepared in Examples 2 to 36 of the present invention

[0262]

[0263]

[0264] Example 38 Cell fluorescence imaging

[0265] The cell fluorescence imaging of the hypocrellin derivatives prepared by the present invention is as follows: Fig.10 As shown. The confocal fluorescence imaging results show that HB-1-PEG16-CH3 (Example 2) has good water solubility and biocompatibility. After incubation with glioma cells (C6 cells), it was found that the photosensitizer can quickly enter the lysosomes of C6 cells and produce good red light fluorescence imaging in the cells, indicating that HB-1-PEG16-CH3 can be used for fluorescence imaging of glioma cells, and the enrichment, distribution and metabolism of drugs in the body can be tracked by fluorescence detection.

[0266] In addition to brain glioma C6 cells, HB-1-PEG16-CH3 can also enter the lysosomes of other cells, such as esophageal cancer cell AKR, gastric cancer cell MFC, lung cancer cell A549, liver cancer cell HCC, bile duct cancer cell MCC, colon cancer cell HCT116, head and neck cancer cell SCC2, brain cancer cell G442, tongue cancer cell TSCa, nasal cancer cell KB, oral cancer cell CAL27, basal cell cancer cell BCC, squamous cell skin cancer cell PECA, melanoma cell B16, prostate cancer cell LNCaP, bladder cancer cell MBT-2. DCFH-DA is used to detect intracellular singlet oxygen. The photosensitizer and fluorescent probe DCFH-DA are co-incubated in the cells. The irradiation time increases to 120s, and the green fluorescence intensity gradually increases, indicating that the singlet oxygen in the cells increases. Other derivatives of the present invention can also perform good cell fluorescence imaging, and the enrichment, distribution and metabolism of drugs in the body can be tracked by fluorescence detection.

[0267] Example 39 Cytotoxicity

[0268] Dark cytotoxicity and phototoxicity experiments were conducted 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, colon cancer cells HCT116 related to digestive tract tumors; head and neck cancer cells SCC2, brain cancer cells G442, tongue cancer cells TSCa, nasal cancer cells KB, oral cancer cells CAL27, brain glioma cells C6 related to head and neck facial tumors; basal cell carcinoma cells BCC, squamous cell skin cancer cells PECA, melanoma cells B16 related to skin tumors; prostate cancer cells LNCaP and bladder cancer cells MBT-2 related to reproductive and urinary systems

[0269] 1) Dark cytotoxicity experiment:

[0270] The cultured tumor cells were added to the culture medium (serum protein and antibiotics), digested with 0.25% trypsin, and pipetted to make a single cell suspension. The cell number was adjusted to about 2x10 4 / mL, 200uL per well was inoculated in a 96-well culture plate and placed in an incubator at 37°C containing 5% CO2 for culture. After the cells adhered to the wall, the supernatant culture medium was discarded, and different concentrations of photosensitizer (commercial HpD, the derivative described in the present invention) were added according to the experimental design under light-proof conditions, and the cells were placed in an incubator for continued culture for 1 hour, and the cell survival rate was detected by the MTT method. 20uL MTT (tetramethyl azo blue) was added to each well, and the culture was terminated after continuing to culture in the incubator for 4 hours, and then 150uL dimethyl sulfoxide was added to each well, and the micro-oscillator was used to shake for 10 minutes to fully dissolve the purple crystals. The wavelength of 570nm was selected, and the optical density value (i.e., OD value) of each well was detected on the microplate reader, and the cell survival rate was calculated: cell survival rate = OD value of the experimental group / OD value of the blank group × 100%.

[0271] like Fig.11 As shown in (a), the cytotoxicity (dark toxicity) research test shows that the cytotoxicity of HB-1-PEG8-CH3 synthesized in Example 2 is relatively small, similar to the commercial photosensitizer hematoporphyrin HpD. Esophageal cancer cell AKR was incubated with photosensitizer at a concentration of 10 μM for half an hour, and no obvious death of AKR cells was observed, indicating that this type of photosensitizer has basically no cytotoxicity.

[0272] The dark toxicity results of some derivatives of the present invention are as follows Figures 11 to 15 As shown in Tables 2 to 4. The hypocrellin derivatives of the present invention are basically non-cytotoxic, and the cell survival rate is above 90% within the concentration range of 20 μM.

[0273] 2) Cell phototoxicity experiment (0.1W / cm 2 ):

[0274] The process of cell phototoxicity experiment was the same as that of dark toxicity experiment. The incubated cells were irradiated with a 635nm semiconductor laser and the power density was adjusted to 0.1W / cm 2 (This light intensity only produces photodynamic effects, but no photothermal effects). The light beam is evenly and vertically irradiated onto the 96-well culture plate for 1000 seconds. Fig.11 The cell phototoxicity study shown in (b) shows that HB-1-PEG8-CH3 exhibits very strong killing power against esophageal cancer cell AKR under red light irradiation. A concentration range of 100nM can kill more than 90% of AKR cells, while the commercial photosensitizer HpD can only kill about 20% of AKR cells under the same conditions, indicating that the photodynamic effect of this type of derivative is significantly better than that of the commercial photosensitizer HpD.

[0275] The phototoxicity test results of some derivatives of the present invention are as follows Figures 11 to 15As shown in Tables 2 to 4. The hypocrellin derivatives of the present invention can kill more than 80 to 90% of tumor cells at a concentration range of 50 nM, and its half lethal concentration IC 50 The value is about 20-30 nM. Therefore, the derivative disclosed by the present invention has better photodynamic effect than the commercial photosensitizer HpD.

[0276] Table 2: MTT data of the hypocrellin derivatives of the present invention on digestive tract tumor cells (0.1W / cm 2 )

[0277]

[0278] Table 3: MTT data of some hypocrellin derivatives of the present invention on head, neck and facial tumor cells (0.1W / cm 2 )

[0279]

[0280] Table 4: MTT data of some hypocrellin derivatives of the present invention on skin tumors and genital urinary tumor cells (0.1W / cm 2 )

[0281]

[0282]

[0283] Fig.11 The dark toxicity graph (a) and phototoxicity graph (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. Fig.11 As shown in (a), the cytotoxicity (dark toxicity) of the photosensitizer indicates that, regardless of whether it has 8 or 16 ethylene glycol units, HB-1-PEG8-CH3 has low cytotoxicity to esophageal cancer AKR cells, and HB-1-PEG16-CH3 has low cytotoxicity to gastric cancer MFC cells, which is similar to the commercial photosensitizer hematoporphyrin HpD. When AKR cells were incubated with 20 μM photosensitizer 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, no obvious death of AKR and MFC cells was observed, indicating that such photosensitizers have basically no cytotoxicity. Fig.11 (b) shows that the photosensitizer exhibited a very strong killing effect on esophageal cancer AKR and gastric cancer MFC cells under red light irradiation. HB-1-PEG8-CH3 at a concentration of 50 nM can kill more than 85% of AKR cells, and the half-lethal concentration IC 50The value (drug concentration required to kill half of the tumor cells) is about 25nM; similarly, 50nM concentration of HB-1-PEG16-CH3 can kill more than 90% of gastric cancer MFC cells, and the half-lethal concentration IC 50 The value is about 25nM; 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 bamboo red fungus polyethylene glycol derivative is significantly better than the commercial photosensitizer hematoporphyrin HpD.

[0284] Fig.12 The dark toxicity graph (a) and phototoxicity graph (b) of the tranexamic acid substituted product of deacetylhypocrellin prepared in Example 7, HC-1-PEG8-CH3, on lung cancer cells A549, and HC-1-PEG16-CH3 on liver cancer HCC cells. Fig.12 As shown in (a), the cytotoxicity (dark toxicity) shows that the cytotoxicity of HC-1-PEG8-CH3 and HC-1-PEG16-CH3 is relatively small, which is similar to the commercial photosensitizer hematoporphyrin HpD. When 20 μM concentration of HC-1-PEG8-CH3 and A549 cells or 20 μM concentration of HC-1-PEG16-CH3 and liver cancer HCC were incubated for half an hour, no obvious death of A549 and HCC cells was observed, indicating that this type of photosensitizer has basically no cytotoxicity. Fig.12 (b) shows that under 635nm laser irradiation, the photosensitizer exhibited very strong killing effects on A549 and HCC cells, respectively. HC-1-PEG8-CH3 at a concentration of 50nM could kill more than 80% of A549 cells, and the half-lethal concentration IC 50 The value is about 30nM; similarly, 50nM concentration of HC-1-PEG16-CH3 can kill more than 90% of HCC cells, and the half-lethal concentration IC 50 The value is about 30nM; 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 bamboo red fungus polyethylene glycol derivative is significantly better than the commercial photosensitizer hematoporphyrin HpD.

[0285] Fig.13 The dark toxicity graph (a) and phototoxicity graph (b) of the hypocrellin B and the glycoside diol amino-polyethylene glycol derivative HB-8-PEG8-CH3 prepared in Example 28 to bile duct cancer MCC cells, and HB-8-PEG16-CH3 to colon cancer HCT116 cells are shown. It can be seen that HB-8-PEG8-CH3 and HB-8-PEG16-CH3 containing long-chain PEG have almost no cytotoxicity under the condition of no light irradiation. After 635nm light irradiation, the concentration range of 50nM can kill more than 90% of bile duct cancer MCC cells, and the half lethal concentration IC50 The value is about 30nM; 50nM concentration can kill more than 90% of colon cancer HCT116, and the half-lethal concentration IC 50 The value is about 30nM; under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 25% of MCC and HCT116 cells, which also shows that the photodynamic effect of HB-8-PEG8-CH3 and HB-8-PEG16-CH3 is significantly better than that of the commercial photosensitizer hematoporphyrin HpD.

[0286] Fig.14 (a) is the phototoxicity graph of HC-8-PEG2-CH3 prepared in Example 28 on brain cancer cells G442 and head and neck cancer cells SCC2. The photosensitizer showed very strong killing power on G442 or SCC2 cells under 635nm laser irradiation. HC-8-PEG2-CH3 at a concentration of 50nM can kill more than 80% of G442 cells, and the half-lethal concentration IC 50 The value is about 20nM; similarly, 50nM concentration of HC-8-PEG8-CH3 can kill more than 90% of SCC2 cells, and the half-lethal concentration IC 50 The value is about 20nM; 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.

[0287] Fig.14 (b) is the phototoxicity graph of the compound HC-8-PEG16-H prepared in Example 29 on tongue cancer cell TSCa and HC-8-PEG32-H on nose cancer cell KB. HC-8-PEG16-H at a concentration of 50 nM can kill more than 90% of tongue cancer cell TSCa, and the half lethal concentration IC 50 The value is about 30nM; while 50nM concentration of HC-8-PEG32-H can kill more than 90% of nasal cancer cells KB, and the half-lethal concentration IC 50 The value is about 30nM; under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 20% of tongue cancer cells TSCa or nasal cancer cells KB.

[0288] Fig.14 (c) Phototoxicity of the compound HC-8-PEG2-SO3H prepared in Example 31 of the present invention to oral cancer cells CAL27 and HC-8-PEG8-SO3H to brain glioma cells C6. HC-8-PEG2-SO3H at a concentration of 50 nM can kill more than 90% of oral cancer cells CAL27, and the half-lethal concentration IC 50The value is about 30nM; while 50nM concentration of HC-8-PEG8-SO3H can kill more than 90% of brain glioma cells C6, and the half-lethal concentration IC 50 The value is about 30nM; under the same conditions, the commercial photosensitizer hematoporphyrin derivative HpD can only kill about 30% of oral cancer cells CAL27 or brain glioma cells C6. The above results show that the photodynamic effect of this type of hypocrellin polyethylene glycol derivative on oral cancer cells CAL27 and brain glioma cells C6 is significantly better than that of the commercial photosensitizer hematoporphyrin HpD.

[0289] Fig.15 (a) is a phototoxicity graph of HB-1-PEG4-H in Example 3, HB-1-PEG16-CO2H in Example 4, and HB-1-PEG24-CO2CH3 in Example 5 on basal cell carcinoma BCC cells; it can be seen from the figure that under the irradiation of red light 635nm, 50nM concentration of HB-1-PEG4-H can kill more than 80% of BCC cells; under the same conditions, 50nM concentration of HB-1-PEG16-CO2H can kill more than 80% of BCC cells, and 50nM concentration of HB-1-PEG24-CO2CH3 can kill more than 85% of BCC cells, and the half lethal concentration IC 50 The values ​​are all about 20 nM, indicating that the above three photosensitizers have a good ability to photodynamically inactivate tumor cells in basal cell carcinoma BCC cells.

[0290] Fig.15 (b) is a phototoxicity graph of HC-1-PEG2-H in Example 8, HC-1-PEG24-SO3H in Example 11, and HB-1-NH-PEG4-CH3 in Example 12 on melanoma B16 cells; it can be seen from the graph that under irradiation with red light of 635nm, HC-1-PEG2-H in a concentration range of 50nM can kill more than 80% of B16 cells; under the same conditions, HC-1-PEG24-SO3H in a concentration range of 50nM can kill more than 85% of B16 cells, and HB-1-NH-PEG4-CH3 in a concentration range of 50nM can kill more than 85% of B16 cells, and their half lethal concentration IC 50 The values ​​are all about 30 nM, indicating that the above three photosensitizers have a good ability to photodynamically inactivate tumor cells in melanoma B16 cells.

[0291] Fig.15(c) is a phototoxicity graph of HB-1-NH-PEG16-H in Example 13, HC-1-NH-PEG4-CH3 in Example 17, and HC-1-NH-PEG24-CO2CH3 in Example 20 on squamous cell carcinoma PECA cells. It can be seen from the figure that under the irradiation of red light at 635nm, HB-1-NH-PEG16-H in the concentration range of 50nM can kill more than 90% of PECA cells; under the same conditions, HC-1-NH-PEG4-CH3 in the concentration range of 50nM can kill more than 85% of PECA cells, and HC-1-NH-PEG24-CO2CH3 in the concentration range of 50nM can kill more than 90% of PECA cells, and its half lethal concentration IC 50 The value is about 20nM, indicating that the above three photosensitizers have a good ability to photodynamically inactivate tumor cells in squamous cell carcinoma PECA cells.

[0292] Fig.15 (d) is a phototoxicity graph 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; it can be seen from the figure that under the irradiation of red light at 635nm, 50nM concentration of HB-2-p-PEG4 can kill more than 85% of LNCaP cells; under the same conditions, 50nM concentration of HB-5-PEG10 can kill more than 85% of LNCaP cells, and 50nM concentration of HB-6-PEG24 can kill more than 90% of LNCaP cells, and the half lethal concentration IC 50 The values ​​are all about 20 nM, indicating that the above three photosensitizers have a good ability to photodynamically inactivate tumor cells in prostate cancer LNCaP cells.

[0293] Fig.15 (e) is a phototoxicity graph of HB-7-PEG4, HC-7-PEG16 in Example 27 of the present invention, and HB-8-PEG32-CO2CH3 in Example 30 on bladder cancer MBT-2 cells; it can be seen from the figure that under the irradiation of red light at 635nm, HB-7-PEG4 in the concentration range of 50nM can kill more than 90% of MBT-2 cells; under the same conditions, HC-7-PEG16 in the concentration range of 50nM can kill more than 90% of MBT-2 cells, and HB-8-PEG32-CO2CH3 in the concentration range of 50nM can kill more than 90% of MBT-2 cells, and their half lethal concentration IC 50 The values ​​are all about 20 nM, indicating that the above three photosensitizers have a good ability to photodynamically inactivate tumor cells in bladder cancer MBT-2 cells.

[0294] Fig.15 (f) is a phototoxicity graph 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. It can be seen from the figure that under the irradiation of red light at 635nm, 50nM concentration of HB-9-PEG16-CH3 can kill more than 80% of BCC cells; 50nM concentration of HB-10-PEG2-CH3 can kill more than 80% of BCC cells, and 50nM concentration of HB-11-PEG16 can kill more than 85% of BCC cells, and the half lethal concentration IC 50 The values ​​are all about 30nM, indicating that the above three photosensitizers have a good ability to photodynamically inactivate tumor cells for basal cell carcinoma BCC.

[0295] In summary, the hypocrellin derivatives of the present invention have almost no cytotoxicity in the absence of light, but have obvious photodynamic effects under light conditions, and under laser irradiation (0.1 W / cm 2 ) can have a strong photodynamic effect. Photosensitizers with a concentration of 10 to 50 nM can kill a variety of tumor cells (such as esophageal cancer cell AKR, gastric cancer cell MFC, lung cancer cell A549, liver cancer cell HCC, bile duct cancer cell MCC, colon cancer cell HCT116, head and neck cancer cell SCC2, brain cancer cell G442, tongue cancer cell TSCa, nasal cancer cell KB, oral cancer cell CAL27, brain glioma cell C6, basal cell cancer cell BCC, squamous cell skin cancer cell PECA, melanoma cell B16, prostate cancer cell LNCaP, bladder cancer cell MBT-2), with a killing rate of more than 85%-90%, and a half-lethal concentration IC 50 The value is about 20-30nM, which is lower than the IC value of commercial photosensitizer hematoporphyrin HpD under the same conditions. 50 The value is 1 to 2 orders of magnitude lower. Therefore, the hypocrellin derivative disclosed in the present invention has better dark toxicity, phototoxicity and photodynamic effect than the commercial photosensitizer hematoporphyrin HpD.

[0296] 3) Cell phototoxicity experiment (0.5W / cm 2 ):

[0297] The above results show that the hypocrellin derivatives prepared by the present invention have good photothermal conversion, and their photothermal conversion efficiency is between 25% and 35% (Table 1). -2 ) can significantly increase the temperature in the environment surrounding the photosensitizer, producing a significant photothermal effect. This shows that the photosensitizer can simultaneously produce singlet oxygen and photothermal effect under appropriate laser irradiation, and can be used for photodynamic / photothermal synergistic therapy.

[0298] For the cell phototoxicity experiment, the incubated cells were irradiated with a 635 nm laser and the power density was adjusted to 0.5 W / cm 2 , so that the light beam is evenly and vertically irradiated onto the 96-well culture plate, and the irradiation time is 1000s. Under this light intensity, there is both photodynamic effect and photothermal effect. Under this condition, killing tumor cells is a synergistic process of photodynamic and photothermal effects. Some derivatives of the present invention are at 0.5W / cm 2 The phototoxicity test results are shown in Table 5. For example, the derivative HB-1-PEG8-CH3 (Example 2) was irradiated with red light (0.5 W / cm 2 ), showing stronger killing power against AKR of esophageal cancer cells. The concentration range of 30nM can kill more than 95% of AKR cells, and the half-lethal concentration (IC50 value) is about 10nM; while at 0.1W / cm 2 Under red light irradiation (other conditions are the same), a concentration of 50nM is required to kill more than 90% of AKR cells, and the half-lethal concentration is about 25nM. Therefore, the synergistic effect of photodynamic and photothermal therapy is significantly better than the effect of photodynamic therapy alone in killing esophageal cancer cells.

[0299] Similarly, the derivative HC-1-PEG16-CH3 (Example 7) was irradiated with red light (0.5 W / cm 2 ), showing stronger killing power against bile duct cancer cells MCC. The concentration range of 30nM can kill more than 95% of MCC cells, and the half-lethal concentration is about 10nM; while at 0.1W / cm 2 Under the irradiation of red light (other conditions are the same), a concentration of 50nM is required to kill more than 90% of MCC cells, and the half-lethal concentration is about 30nM. It also shows that the synergistic effect of photodynamic and photothermal therapy is significantly better than the effect of photodynamic therapy alone in killing cholangiocarcinoma tumor cells.

[0300] Another example: The derivative HB-1-NH-PEG8-CH3 (Example 12) was irradiated with red light (0.5 W / cm 2 ), showing stronger killing power against brain glioma cells C6. The concentration range of 30nM can kill more than 95% of C6 cells, and the half-lethal concentration is about 10nM; 2 Under the irradiation of red light (other conditions are the same), a concentration of 50nM is required to kill more than 85% of C6 cells, and the half-lethal concentration is about 30nM. It also shows that the synergistic effect of photodynamic and photothermal killing of brain glioma cells is significantly better than the effect of photodynamic alone.

[0301] Another example: The derivative HC-1-NH-PEG16-CH3 (Example 17) was irradiated with red light (0.5 W / cm 2), showing stronger killing power against basal cell carcinoma BCC cells. The concentration range of 30nM can kill more than 95% of BCC cells, and the half-lethal concentration is about 10nM; while at 0.1W / cm 2 Under red light irradiation (0.5W / cm 2 )(other conditions being the same), a concentration of 50 nM is required to kill more than 90% of BCC cells, and the half-lethal concentration is about 25 nM. This also shows that the synergistic effect of photodynamic and photothermal therapy is significantly better than the effect of photodynamic therapy alone in killing basal cell cancer cells.

[0302] The above results show that the hypocrellin derivatives prepared by the present invention have good photothermal conversion efficiency. -2 ) can significantly increase the temperature in the environment around the photosensitizer, producing a significant photothermal effect. This shows that the photosensitizer can simultaneously produce singlet oxygen and photothermal effect under appropriate laser irradiation, and can be used for photodynamic / photothermal synergistic therapy. Table 5 also lists other derivatives at 0.5 W / cm 2 MTT results of killing various tumor cells under light. It can be seen that this type of derivative can kill more than 90-95% of tumor cells in the concentration range of 30nM, and its half lethal concentration IC 50 The value is about 10-15 nM. Therefore, the derivatives of the present invention kill various tumor cells under the synergistic effect of photodynamics and photothermals, which is significantly better than the effect of photodynamics alone.

[0303] Table 5: MTT data of the derivatives of the present invention on digestive tract tumors, head and neck facial tumors, skin tumors and genital urinary tumor cells (0.5W / cm 2 )

[0304]

[0305] Example 40 In vivo enrichment of photosensitizing drugs

[0306] In order to study the enrichment process of the hypocrellin derivative prepared by the present invention in subcutaneous tumors, this experiment used mice as a model, inoculated subcutaneous tumors with esophageal cancer AKR cells, and obtained an esophageal cancer subcutaneous mouse tumor model. The derivative HB-1-PEG16-CH3 in Example 2 of the present invention was used as a photosensitizer and injected into tumor-bearing mice through the tail vein at a dose of 10 mg / kg. Its fluorescence imaging behavior in the esophageal cancer AKR cell tumor-bearing mice was observed, and the fluorescence signal of the tumor site at 4 hours was collected using a multi-spectral small animal in vivo imaging system. The animal fluorescence imaging is as follows: Fig.17As shown in (a), it can be seen that after several cycles in the body via intravenous injection, the drug molecule HB-1-PEG16-CH3 has a strong enrichment ability in the tumor through passive targeting for about 4 hours.

[0307] Then, lung cancer A549 cells were inoculated into subcutaneous tumors to obtain a lung cancer subcutaneous mouse tumor model. The derivative HC-1-PEG16-CH3 in Example 7 of the present invention was used as a photosensitizer 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, and the fluorescence signal of the tumor site at 4 hours was collected using a multi-spectral small animal in vivo imaging system. The animal fluorescence imaging is as follows: Fig.17 As shown in (b), it can be seen that after several cycles in the body via intravenous injection, the drug molecule HC-1-PEG16-CH3 has a strong enrichment ability in the tumor through passive targeting for about 4 hours.

[0308] The above results show that the hypocrellin derivatives provided in the present invention can be administered to tumor-bearing mice by tail vein injection. From the in vivo imaging of small animals, it can be seen that these drugs produce significant fluorescent signals at the tumor of the mouse, indicating that such derivatives can be used as photosensitizers to have a good enrichment effect at the tumor of 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, brain glioma, basal cell carcinoma, squamous cell skin cancer, melanoma, prostate cancer, and bladder cancer.

[0309] The above is an explanation of the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A hypocrellin derivative represented by formula (I), and its isomers, isotope-labeled substances or pharmaceutically acceptable salts: In formula (I), the substituent R1 is -H or -COCH3; The general structural formula of the substituent R2 is -NH-(CH2) p —X—Y—(CH2CH2O) n —Z; formula (II); in, X is a linking group selected from cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl; Y is a linking group selected from the following groups: -C(=O)O-, -C(=O)O-CH2-, -OC(=O)-, -OC(=O)-CH2-, -OC(=O)-CH2O-, -C(=O)NH-, the left side of the above group is connected to X, and the right side is connected to the polyethylene glycol unit; Z is a terminal group selected from -H, -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; -(CH2CH2O) n - is a polyethylene glycol unit, and n is an integer selected from 1 to 100.

2. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: Z is selected from -H, -CH3, -C2H5, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H.

3. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: When X is a cyclohexyl group, the group -(CH2) connected to the cyclohexyl group p - and group Y are located in the para, ortho or meta position; When X is a cyclopentyl group, the group -(CH2) connected to the cyclopentyl group p - and group Y are located in the ortho or meta position.

4. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: R2 is selected from: -NH-CH2C6H 10 -COO-(CH2CH2O) n -Z、-NH-CH2C6H 10 -COO-CH2-(CH2CH2O) n -Z、-NH-CH2C6H 10 -CO-NH-(CH2CH2O) n -Z、-NH-CH2C6H 10 -O-CO-(CH2CH2O) n -Z、-NH-C6H 10 -COO-(CH2CH2O) n -Z、-NH-C6H 10 -CO-NH-(CH2CH2O) n -Z、-NH-C6H 10 -COO-CH2-(CH2CH2O) n -Z、-NH-C6H 10 -O-CO-(CH2CH2O) n -Z、-NH-C6H 10 -O-CO-CH2-(CH2CH2O) n -Z、-NH-C6H 10 -O-CO-CH2O-(CH2CH2O) n -Z、-NH-C5H8-COO-(CH2CH2O) n -Z、-NH-C5H8-CO-NH-(CH2CH2O) n -Z、-NH-C5H8-COO-CH2-(CH2CH2O) n -Z、-NH-C5H8-O-CO-(CH2CH2O) n -Z、-NH-C5H8-O-CO-CH2-(CH2CH2O) n -Z、-NH-C5H8-O-CO-CH2O-(CH2CH2O) n -Z、-NH-C4H6-COO-(CH2CH2O) n -Z、-NH-C4H6-CO-NH-(CH2CH2O) n -Z、-NH-C4H6-COO-CH2-(CH2CH2O) n -Z、-NH-C4H6-O-CO-(CH2CH2O) n -Z、-NH-C4H6-O-CO-CH2-(CH2CH2O) n -Z、-NH-C4H6-O-CO-CH2O-(CH2CH2O) n -Z、-NH-C3H4-COO-(CH2CH2O) n -Z、-NH-C3H4-CO-NH-(CH2CH2O) n -Z、-NH-C3H4-COO-CH2-(CH2CH2O) n -Z、-NH-C3H4-O-CO-(CH2CH2O) n -Z、-NH-C3H4-O-CO-CH2-(CH2CH2O) n -Z、-NH-C3H4-O-CO-CH2O-(CH2CH2O) n -Z、 Each n is the same or different and is independently selected from an integer of 1 to 100; Z is selected from -H, -CH3, -C2H5, -CH2COOH, -CH2CH2COOH, -CH2CH2COOCH3 or -CH2CH2SO3H; -C6H 10 - represents cyclohexyl, -C5H8- represents cyclopentyl, -C4H6- represents cyclobutyl, and -C3H4- represents cyclopropyl.

5. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: The R2 is selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Or selected from: Wherein, each n is the same or different and is independently selected from integers of 1 to 100.

6. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: The R2 is selected from Or selected from 7. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: The structural formula of the hypocrellin derivative is:

8. The derivative according to claim 1, and its isomers, isotope-labeled substances or pharmaceutically acceptable salts, characterized in that: The general structural formula of the hypocrellin derivatives of formula (I) above also includes the enol tautomer shown in formula (I'):

9. A pharmaceutical composition comprising the hypocrellin derivative according to any one of claims 1 to 8, and at least one of its isomers, isotope labels or pharmaceutically acceptable salts; the pharmaceutical composition is used as a photodynamic therapy drug and / or a fluorescence-mediated drug and / or a photothermal therapy drug.

10. Use of the hypocrellin derivative according to any one of claims 1 to 8, or at least one of its isomers, isotope labels or pharmaceutically acceptable salts in the preparation of anti-tumor drugs.

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

12. The use according to claim 10, characterized in that The tumors are digestive tract tumors, head and neck facial tumors, skin tumors, and genital and urinary 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; the genital and urinary system tumors include prostate cancer and bladder cancer.

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