WO2018086241A1 - Ph-sensitive 1,4-disubstituted zinc phthalocyanine coordination complex, preparation method therefore, and application thereof in medicine - Google Patents

Abstract

The present invention relates to a pH-sensitive ketal-linked cholesterol-zinc phthalocyanine coordination complex and a preparation method therefor and application thereof in medicine. In particular, the present invention relates to the zinc phthalocyanine coordination complex shown in a general formula (I), a preparation method therefor, a pharmaceutical composition containing the coordination complex, and use thereof as a photosensitizer, especially use in treating cancers, wherein all substituents in the general formula (I) have the same definition as those in the description. As cholesterol radicals exist, the series of compounds are difficult for tumor cells and normal cells to take in; however, in an external subacid environment of tumor tissue cells, hydrolysis reaction of ketal occurs, and hydrolysis derivatives of zinc phthalocyanine can be easily taken in by cancer cells and show extremely-high photosensitive activity. The zinc phthalocyanine coordination complexes can be prepared into tumor cell external subacid environment target photosensitive medicine.

Description

pH-sensitive 1,4-disubstituted phthalocyanine zinc complex, preparation method thereof and application in medicine Technical field

The invention belongs to the field of medicine, relates to zinc phthalocyanine complexes, a preparation method thereof and application thereof in medicine, and discloses the use thereof as a photosensitizer for treating cancer.

technical background

Photodynamic Therapy (PDT), also known as Photoradiation Therapy (PRT) or Photochemotherapy, is a treatment based on the principle of photochemical reaction of specific chemicals. The chemical used is called a tumor chemistry drug (also known as photosensitizer, PS for short). In the PDT therapy, the photosensitizer is injected into the body by intravenous injection (for the skin, it can also be applied to the affected area). After a certain period of time, the tumor tissue is irradiated with light of a specific wavelength, and the photosensitizer enriched in the tumor tissue is excited by light. , produces a series of photophysical chemical reactions that produce cytotoxic reactive oxygen species, thereby killing cancer cells and destroying tumor tissue.

In 1996, it was approved by the US FDA for clinical use. In 1997, the FDA listed it as one of the five basic methods of cancer treatment (surgery, radiotherapy, chemotherapy, photodynamic, biochemical immunity). Compared with traditional therapies, PDT has minimal trauma, low toxicity, good selectivity, good applicability, reproducible treatment, palliative treatment, synergistic surgery to improve curative effect, elimination of recessive cancer lesions, protection of appearance and Important organ function, short treatment time and other advantages.

Photodynamic therapy can also effectively treat non-cancer diseases such as bacterial infections, oral infections, macular degeneration, arteriosclerosis, traumatic infections, and skin diseases. Photosensitizers can also be used for photodynamic disinfection, most importantly for sterilization of blood and blood derivatives. At the same time, photodynamic diagnosis using the fluorescent properties of photosensitizers is also an important use of medical photosensitizers.

The key to photodynamic therapy lies in photosensitizers, and the photodynamic efficacy depends on the pros and cons of photosensitizers. Based on the potential of photodynamic therapy in the treatment of tumors and other diseases, the scientific community generally believes that photodynamic therapy will become an important medical method in the 21st century. The main photosensitizer used in clinical practice is Porfimer sodium (Photofin), which has been listed in 28 countries and regions such as the Netherlands, Canada, Japan, the United States, France, Germany, and the United Kingdom. China's research on photodynamic therapy (PDT) started later than the United States, Japan and other countries, but the progress is faster. Since the early 1980s, a lot of work has been done to develop photodynamic therapy. Compared with the results reported abroad, there is not much difference between its depth and breadth. Chongqing Huading Modern Bio-Pharmaceutical Co., Ltd. has developed and produced Hematoporphyrin (Hematoporphyrin), which has been officially approved by the State Food and Drug Administration (SFDA) for production, sales and clinical application. Xibo is the only anti-cancer photosensitizer listed in China. It is a foreign-listed anti-cancer analog, both of which are hematoporphyrin derivatives, and belong to the first generation of anti-cancer photosensitive drugs.

Despite the clinical success of 卟非姆纳 and 喜泊分, its components are complex, and the role of various components in photodynamic damage has not been clarified so far, and the inactive ingredients accounting for more than 20% of the total amount of drugs are not only It can not produce effective photodynamic damage to the target tissue of the lesion, but instead becomes the culprit leading to photosensitive reaction in normal tissues. In addition, the first generation of photosensitizing drugs have a weak targeting effect on tumors. It is a recent research hotspot to study high-efficiency and low-toxic photosensitizing drugs with tumor targeting.

The hypoxic microenvironment in the solid tissue of the tumor results in a lower pH outside the tumor (about 6.5), while the extracellular pH of the normal tissue is about 7.4. The difference in pH between tumor solid tissue and normal tissue provides a new strategy for the design of tumor-targeted drugs. The inventors of the present invention have reported in recent years a number of series of pH-sensitive photosensitive compounds, such as axial amino derivatives substituted silicon phthalocyanine, which affect the photosensitivity by different degrees of ionization of amino groups in different pH environments ( Jiang, X.-J .et.al, Chem.Commun., 2010, 46, 3188-3190); Axial phenyl derivatives replace silicon phthalocyanine, which affects photosensitivity by different degrees of aggregation of phthalocyanine molecules in a pH environment ( Jiang, X.- J. et . al, Chem. Eur. J. , 2010, 16, 4777-4783); fluoroboron dipyrrole derivatives and ferrocene derivatives linked by pH-sensitive ketal linkages, due to ferrocene fluorescence The quencher can quench the photoactive activity of the fluoroboron dipyrrole derivative by photoinduced electron transfer (PET) process, but in the micro acid environment, the ketal is hydrolyzed and the ferrocene is supplied with electrons. The group is detached and the photosensitivity of the dipyrrole derivative is restored ( Jiang, X.-J. et. al, Chem. Eur . J. 2016, 22, 8273-8281). However, these compounds and most of the pH-sensitive drugs reported so far are not tumor cell extracellular micro-acid environment-targeting drugs, because the subcellular tissues of tumor cells and normal cells such as mitochondria and lysosomes are acidic, lysozyme The pH of the body can be as low as 5. These compounds, when taken up by tumor cells and normal cells, can be stimulated by the acid in the subcellular tissue to damage the photosensitizing activity and damage the tumor cells and normal cells.

Summary of the invention

The present invention discloses a series of tumor extracellular microacid environments that target photosensitizing drugs. The structure, synthesis and application of a series of ketal-linked zinc phthalocyanine complexes and cholesterol derivatives are disclosed. Introducing a cholesterol group at the 1,4 position of zinc phthalocyanine, an acid-sensitive ketal bond between zinc phthalocyanine complex and cholesterol, zinc phthalocyanine-cholesterol complex due to the non-polar nature of cholesterol and large steric hindrance The substance is difficult to be taken up by cancer cells and normal cells, but in the micro-acid environment outside the tumor solid tissue, the ketal bond is hydrolyzed, the cholesterol group is shed, and the zinc phthalocyanine hydrolyzed derivative can be taken up by the cancer cells and exhibited. Extremely high photosensitivity, they are tumor-specific extracellular micro-acid environments that target photosensitizing drugs.

The present invention provides a compound of the formula (I):

Wherein: n = 1, 2, 3, or 4; X = O or NH;

Or a pharmaceutically acceptable salt thereof.

Chemical formula (1) for hydrolysis of a compound of the formula (I) in a slightly acidic environment

The hydrolysate of the compound of the formula (I) is a compound of the formula (V) having the same structure and similar structure as the compound reported in the literature (Liu, J.-Y., Jiang, X.-J.et. Al., Org. Biomol. Chem., 2008, 6, 4560-4566; Liu, J.-Y., lo, P.-C., Jiang, X.-J., et . al., Dalton Trans. , 2009, 4129-4135), these compounds have high uptake rates in tumor cells and exhibit very high photosensitivity at very low concentrations.

Typical compounds of the invention include, but are not limited to:

Or a pharmaceutically acceptable salt thereof, wherein

The present invention also provides a process for the preparation of a compound of the formula (I), which comprises:

Wherein: n = 1, 2, 3, or 4; X = O or NH;

In the first step, the solvent is selected from the group consisting of N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, chloroform and tetrahydrofuran; the reaction is carried out at a temperature of -5 to 80 ° C; The basic condition is provided by a reagent selected from the group consisting of pyridine, triethylamine, sodium hydride and 4-N,N-lutidine; the molar ratio of the compound of the formula (II) to cholesterol formyl chloride is 1:0.2-2 .

In the second step, the solvent is selected from the group consisting of N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, chloroform and tetrahydrofuran; the reaction is carried out at a temperature of -5 to 80 ° C; The basic condition is provided by a reagent selected from the group consisting of pyridine, triethylamine, sodium hydride and 4-N,N-lutidine; the molar ratio of the compound of the formula (IV) to the compound of the formula (III) is 1: 0.5 to 5.

If necessary, the compound can also be purified by methods well known to those skilled in the art, such as by distillation, by silica gel column chromatography or by high performance liquid chromatography (HPLC).

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound of the formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

The present invention also relates to the use of a compound of the formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for the preparation of a photodynamic drug or a photosensitizing drug.

The present invention also relates to a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof, or a medicament comprising the same Use of a composition for the manufacture of a medicament for the treatment of cancer. The cancer described therein is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, breast cancer, bladder cancer, prostate cancer, pancreatic cancer, cholangiocarcinoma, rectal cancer, colon cancer, skin cancer, head and neck cancer, Eye tumor, uterine cancer and ovarian cancer, excellent breast cancer.

The present invention also relates to a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, which is used as a photodynamic drug or a photosensitizing drug.

The present invention also relates to a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, which is useful for treating cancer. The cancer described therein is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, breast cancer, bladder cancer, prostate cancer, pancreatic cancer, cholangiocarcinoma, rectal cancer, colon cancer, skin cancer, head and neck cancer, eye tumor, uterine cancer and ovary. Cancer, preferably breast cancer.

The present invention also relates to a method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, and then suitably The light source is illuminated. The suitable light source may be provided by a conventional light source coupled to a suitable filter or by a laser of a particular wavelength, the source having a wavelength in the range of 550 to 900 nm, preferably 620 to 720 nm.

The compounds according to the invention may be administered orally, sublingually, parenterally, subcutaneously, intramuscularly, intravenously, transdermally, topically or rectally.

In the pharmaceutical compound of the present invention, for oral administration, sublingual administration, parenteral administration, subcutaneous administration, intramuscular administration, intravenous administration, transdermal administration, topical administration or rectal administration, the active ingredient may be conventionally used. The pharmaceutically acceptable carriers are mixed together and administered to the animal or human in the form of an administration unit. Suitable administration unit forms include oral forms such as tablets, gel capsules, powders, granules and solutions or suspensions for oral administration, sublingual or buccal administration, parenteral, subcutaneous, intramuscular, intravenous, nasal Internal or intraocular administration forms and rectal administration forms.

When the solid composition is prepared in the form of a tablet, the main active ingredient is mixed with a pharmaceutically acceptable carrier such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic or the like. The tablets may be coated with sucrose or other suitable materials or treated in such a way that they have prolonged or delayed activity and continuously release a predetermined amount of active ingredient.

A gel capsule preparation is obtained by mixing the active ingredient with a diluent and by pouring the obtained mixture into a soft or hard capsule.

Formulations in the form of syrups or elixirs may contain the active ingredient along with sweetening agents, preservatives, and perfuses, and suitable colorants.

The powder or granules which may be dispersed in water may contain the active ingredient in admixture with a dispersing agent, a surfactant, a wetting or suspending agent, and a flavoring or sweetening agent. The pharmaceutical composition contains polyoxyethylene castor oil and its derivatives, dimethyl sulfoxide, ethanol, glycerin, N, N-dimethylformamide, polyethylene glycol 300-3000, cyclodextrin, glucose, Tween One or more of polyethylene glycol monostearate.

Suppositories are used for rectal administration, which are prepared using a binder that melts at the rectal temperature, for example, cocoa butter or polyethylene glycol.

An aqueous suspension, an isotonic physiological saline solution or a sterile and injectable solution (including a medicament) A physiologically compatible dispersing and/or wetting agent) is for parenteral, intranasal or intraocular administration. The pharmaceutical composition contains polyoxyethylene castor oil and its derivatives, dimethyl sulfoxide, ethanol, glycerin, N, N-dimethylformamide, polyethylene glycol 300-3000, cyclodextrin, glucose, Tween One or more of polyethylene glycol monostearate.

The active ingredient (possibly together with one or more additive carriers) can also be formulated as a microcapsule.

The compounds of the invention can be administered at doses between 0.01 mg/day and 5000 mg/day, in a single dose/day manner or in several doses throughout the day, for example, the same dose twice daily. . The daily dose administered is advantageously between 0.1 mg and 200 mg, even more advantageously between 2.5 mg and 50 mg. It may be desirable to use dosages outside of these ranges, as will be appreciated by those skilled in the art.

In a particular embodiment of the invention, the pharmaceutical composition may also be formulated for external administration. It can be introduced into the usual form of the application type (i.e., especially lotions, foams, gels, dispersants, sprays), the usual forms having excipients, in particular excipients It is able to penetrate the skin in order to improve the properties and accessibility of the active ingredients. In addition to the compositions according to the invention, these compositions generally further comprise a physiologically acceptable medium, which usually comprises water or a solvent, for example an alcohol, an ether or an ethylene glycol. The composition may further comprise a surfactant, a preservative, a stabilizer, an emulsifier, a thickener, other active ingredients that produce a complementary effect or a possible synergistic effect, trace elements, essential oils, perfumes, colorants, collagen, Chemical or mineral filter.

definition

Unless otherwise stated, the following terms used in the specification and claims have the following meanings.

In the present invention, "pharmaceutically acceptable" is understood to mean that it is used in the preparation of a pharmaceutical composition which is generally safe, non-toxic, biologically or otherwise satisfying needs and said combination Objects can be accepted for use in mammals and humans.

In the present invention, a "pharmaceutically acceptable salt" of a compound is understood to mean a salt which is a pharmaceutically acceptable (as defined herein) salt and which possesses the desired pharmacological activity of the parent compound. This salt includes:

(1) Acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or with organic acids such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid , fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, sticky An acid addition salt formed by an acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, dibenzoyl-L-tartaric acid, tartaric acid, p-toluenesulfonic acid, trimethylacetic acid, trifluoroacetic acid or the like; with

(2) When the acid proton present in the parent compound is replaced by a metal ion such as an alkali metal ion (for example, Na ⁺ , K ⁺ or Li ⁺ ), an alkaline earth metal ion (such as Ca ²⁺ or Mg ²⁺ ) or aluminum ion. Or a salt formed when coordinated with an organic or inorganic base. Acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, and the like. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, and sodium hydroxide.

"Pharmaceutical composition" means a mixture comprising one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers. And excipients. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby exerts biological activity.

"Ts" is p-toluenesulfonate.

detailed description

Those skilled in the art will better understand the present invention by reading the following examples. These examples are only intended to explain the invention.

The experimental methods in the examples of the present invention which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the raw material or the manufacturer of the commodity. Reagents without specific source are routine reagents purchased from the market. The reference compound method of the formula (IV) is easily synthesized (New J. Chem., 2013, 37, 1746-1752; Org. Biomol. Chem., 2008, 6, 4560-4566; Dalton Trans., 2009, 4129 -4135).

Nuclear Magnetic Resonance Instrument: Bruker ARX-300 high resolution high resolution nuclear magnetic resonance instrument.

Mass Spectrometry: QSTAR Elite tandem quadrupole time-of-flight mass spectrometer.

MTT Tester: Thermo Scientific Multiskan GO Full Wavelength Microplate Reader

PBS buffer: phosphate buffer

The structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). The NMR chemical shift (δ) is given in units of 10 ^-6 (ppm). The solvent was determined to be deuterated dimethyl sulfoxide (DMSO-d ₆ ), and the internal standard was tetramethylsilane (TMS). The following abbreviations are used: s is a single peak, bs is a broad single peak, d is a doublet, t is a triplet, qdt is a quartet, m is a multiple or a large number of peaks, and dd is a doublet.

The thin layer chromatography silica gel plate uses Qingdao GF254 silica gel plate, and the silica gel plate used for thin layer chromatography (TLC) has a specification of 0.15 mm to 0.2 mm, and the thin layer chromatography separation and purification product adopts a specification of 0.4 mm to 0.5 mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.

In the examples, unless otherwise specified, the reactions were all carried out under an argon atmosphere or a nitrogen atmosphere.

In the examples, the solution in the reaction means an aqueous solution unless otherwise specified.

In the examples, the temperature of the reaction was room temperature unless otherwise specified.

The monitoring of the progress of the reaction in the examples was carried out by thin layer chromatography (TLC).

Synthesis of Compound 1 of Example 1

step 1

In an ice water bath, compound 1-1 (1.62 g, 10 mmol), cholesteryl chloride (2.2 g, 4.9 mmol) and triethylamine (2 g, 20 mmol) were added to dichloromethane (50 mL), and the mixture was stirred for 2 hr. After the reaction, water (100 mL) was added to the reaction mixture, and the mixture was stirred and the organic phase was collected. The organic phase was dried over anhydrous sodium sulfate and evaporated under reduced pressure. The crude product was purified by silica gel chromatography eluting with chloroform. /methanol (9:1) gave 1-2 (1.53 g, 55%) as a white solid. MS (ESI): m/z = 598 [M+Na] ⁺ .

Step 2

Compound 1-3 (1.27 g, 1 mmol), 1-2 (1.55 g, 2.7 mmol) and pyridine (0.8 g, 10.1 mmol) were added to chloroform (25 mL). The reaction was stopped, water (120 mL) was added to the reaction mixture, and the mixture was stirred and the organic phase was collected. The organic phase was dried over anhydrous sodium sulfate and evaporated under reduced pressure. The crude product was purified by silica gel chromatography. Methane/methanol (20:1) gave a green solid 1 (1. 18 g, 57%). ^{1 H NMR (300MHz, DMSO-} d 6): δ9.33-9.37 (m, 4H, Pc-H α), 9.21-9.25 (m, 2H, Pc-H α), 8.15-8.20 (m, 6H, Pc-H _β ), 7.53 (s, 2H, Pc-H _β ), 5.28-5.37 (m, 2H, chol-6-CH), 4.82-4.90 (m, 4H, CH ₂ ), 4.35-4.49 (m , 6H, CH ₂ , chol-3-CH), 3.85-4.10 (m, 8H, CH ₂ ), 3.41-3.75 (m, 32H, CH ₂ ), 1.72-2.35 (m, 16H, chol), 0.85- 1.58 (s, 76H, chol, CH ₃ ), 0.68 (s, 6H, chol-18-CH ₃ ). MS (ESI): m / z = 2077 [M + H] +.

Synthesis of Compound 2 of Example 2

step 1

In an ice water bath, compound 2-1 (1.64 g, 10 mmol), cholesteryl chloride (1.55 g, 3.5 mmol) and pyridine (0.8 g, 10.1 mmol) were added to dichloromethane (25 mL). The reaction was stopped for 2 hours, the reaction was stopped, water (100 mL) was added to the reaction mixture, and the mixture was stirred. The organic phase was collected. The organic phase was dried over anhydrous sodium sulfate and evaporated under reduced pressure. The solvent was chloroform/methanol (15:1) to afford white crystals 2-2 (1.02 g, 49%). MS (ESI): m/z = 495 [M+Na] ⁺ .

Step 2

Compound 2-3 (0.7 g, 0.6 mmol), 2-2 (1.6 g, 2.8 mmol) was added to tetrahydrofuran (30 mL), and sodium hydride (0.1 g, 4.2 mmol) was slowly added to the reaction mixture. Stirring the reaction for 2 hours, slowly adding 2 ml of water to quench the reaction, adding water (100 mL) to the reaction mixture, stirring and standing, collecting the organic phase, the organic phase was dried over anhydrous sodium sulfate, and evaporated. The column was separated and purified, and the eluent was chloroform/methanol (20:1) to afford a blue solid 2 (0.25 g, 22%). ^{1 H NMR (300MHz, DMSO-} d 6): δ9.32-9.36 (m, 4H, Pc-H α), 9.20-9.25 (m, 2H, Pc-H α), 8.12-8.2 (m, 6H, Pc-H _β ), 7.51 (s, 2H, Pc-H _β ), 5.30-5.37 (m, 2H, chol-6-CH), 4.81-4.91 (m, 4H, CH ₂ ), 4.32-4.48 (m , 10H, CH ₂ , chol-3-CH), 3.85-4.12 (m, 8H, CH ₂ ), 3.41-3.75 (m, 20H, CH ₂ ), 1.70-2.35 (m, 16H, chol), 0.83- 1.57 (s, 76H, chol, CH ₃ ), 0.67 (s, 6H, chol-18-CH ₃ ). MS (ESI): m / z = 1992 [M] +.

Synthesis of Compound 3 of Example 3

In an ice water bath, compound 3-3 (0.6 g, 5.5 mmol), 2-2 (0.8 g, 1.4 mmol) was added to tetrahydrofuran (30 mL), and sodium hydride (0.1 g, 4.2 mmol) was slowly added to the reaction mixture. Stirring the reaction for 2 hours, slowly adding 2 ml of water to quench the reaction, adding water (100 mL) to the reaction mixture, stirring and standing, collecting the organic phase, the organic phase was dried over anhydrous sodium sulfate, and evaporated. The column was separated and purified, and the eluent was trichloromethane/methanol (20:1) to afford a blue solid 3 (0.22 g, 21%). ¹ H NMR (300MHz, DMSO-d ₆ ): δ 9.35-9.49 (m, 6H, Pc-H _α ), 8.12-8.2 (m, 6H, Pc-H _β ), 7.58 (s, 2H, Pc- H _β ), 5.31-5.35 (m, 2H, chol-6-CH), 4.81-4.98 (m, 4H, CH ₂ ), 4.32-4.56 (m, 10H, CH ₂ , chol-3-CH), 4.10 -4.20 (m, 4H, CH ₂ ), 3.41-3.75 (m, 12H, CH ₂ ), 1.70-2.35 (m, 16H, chol), 0.80-1.62 (s, 76H, chol, CH ₃ ), 0.66 ( s, 6H, chol-18-CH ₃ ). MS (ESI): m/z = 1904 [M] ⁺ .

Synthesis of Compound 4 of Example 4

Compound 4-3 (0.7 g, 0.7 mmol), 1-2 (1.33 g, 2.1 mmol) and triethylamine (2 g, 20 mmol) were added to chloroform (30 mL). The reaction was stopped, water (100 mL) was added to the reaction mixture, and the mixture was stirred and allowed to stand. The organic phase was collected. The organic phase was dried over anhydrous sodium sulfate and evaporated under reduced pressure. Methyl chloride/methanol (20:1) gave a blue solid 4 (0.25 g, 20%). ^{1 H NMR (300MHz, DMSO-} d 6): δ9.33-9.53 (m, 6H, Pc-H α), 8.10-8.18 (m, 6H, Pc-H β), 7.58 (s, 2H, Pc- H _β ), 5.36-5.40 (m, 2H, chol-6-CH), 4.73-4.80 (m, 4H, CH ₂ ), 4.38-4.60 (m, 2H, chol-3-CH), 3.35-3.85 ( m, 20H, CH ₂ ), 1.73-2.40 (m, 16H, chol), 0.84-1.65 (s, 76H, chol, CH ₃ ), 0.68 (s, 6H, chol-18-CH ₃ ). MS (ESI): m/z = </RTI> [M+H] ⁺ .

Test case: in vitro anti-tumor cell light-sensitive experiment

Test article: Compound 2 of the present invention

Positive control: hematoporphyrin injection (English name: Hematoporphyrin Injection; trade name: Xibo, produced by Chongqing Huading Modern Biopharmaceutical Co., Ltd.).

Test cells: human breast cancer cells MCF-7

Main reagents: 1) RPMI-1640 complete medium: Add 500,000 U of penicillin/streptomycin, 56 mL of fetal bovine serum to 500 mL of RPMI-1640 liquid medium (GIBCO), and mix. 2) MTT solution (MTT: 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide, purchased from MP Company, USA): 5 mg/mL of powdered MTT The concentration is dissolved in PBS solution, sterilized by filtration, and is now ready for use.

experimental method:

1) Preparation method of the test sample: the test sample is formulated into a mother liquor having a concentration of 1 mM in DMSO; 100 μL of a 1 mg/mL mother liquor is taken during the experiment, and 1.15 mL of 0.5% (w/w) polyoxyethylene castor oil pH 7.4 PBS is added. pH 6.5 PBS buffer, formulated into 80μg / mL drug solution, and diluted with the corresponding PBS buffer into different concentrations of the drug solution, the pH value of the drug solution is kept unchanged during the dilution process, and the drug solution is cultured immediately after preparation. . The final concentration of DMSO in each drug and negative control group was < 1%. Xibofen is a 25 mL preparation containing 5 mg of liquid solution at a concentration of 5 mg/mL. Take 100 μL of 5 mg/mL preparation, add 4.90 mL of pH 7.4 PBS or pH 6.5 PBS buffer, and dilute to different concentrations of the solution with the corresponding PBS buffer. Keep the pH of the solution unchanged during the dilution process. Cell dosing culture is performed immediately.

2) The adherent tumor cells in logarithmic growth phase were selected and digested with trypsin, and the cell suspension containing the appropriate concentration was prepared by using RPMI 1640 medium containing 10% fetal bovine serum, and inoculated into a 96-well culture plate. Inoculate 100 μL per well. Shake the cell suspension after each row. After adding the cells, gently rotate the plate horizontally to spread the cells evenly on the surface of the well. Add a sterile PBS around the well of the 96-well plate. Incubate for 24 hours at ° C, 5% CO ₂ . Then, 100 μL of each test drug, positive drug, solvent and culture solution of different concentrations were added, and 3 parallel holes were set in each group. After mixing, it was divided into two groups: light and dark. After co-cultivation for 2 hours, the medium was discarded, and the medium containing no test sample was added again. The culture was continued at 37 ° C and 5% CO ₂ . hour. After 24 hours, add 5 mg/mL MTT per well, 20 μL, and incubate for 4 hours at 37 ° C, 5% CO ₂ , carefully aspirate the supernatant, add 200 μL of DMSO to each well, and shake for 10 minutes to form the formed formazan. After the particles were sufficiently dissolved, the absorbance was measured by a microplate reader, and the measurement wavelength was 570 nm, and the reference wavelength was 630 nm. The light source is connected to the insulated water tank by a 200W halogen lamp and a filter larger than 610 nm, and the light dose is 48 J cm ^-2 .

3) Calculation method of drug inhibition rate of tumor cell growth: tumor cell growth inhibition rate (%) = [(negative control group OD mean - administration group OD mean) / negative control group OD mean value] × 100%. The calculation of the half-inhibitory concentration IC ₅₀ was determined by logit regression.

Experimental results:

Table 1 IC ₅₀ (ng/mL) values of compound 2 and Xipo fraction in human breast cancer cell MCF-7 at illumination

Photosensitive drug	Cell pH 7.4 liquid culture	Cell pH 6.5 liquid culture
Compound 2	>8000	95
Hi bo	3800	4100

The experimental results showed that all the tested compounds showed no cytotoxicity at concentrations up to 5000 ng/mL in the dark, but the human breast cancer cells MCF-7 at pH 6.5 and pH 7.4 under illumination. the lethal concentration IC ₅₀ value or less, between 3800-4100ng / mL. However, when Compound 2 was cultured in a pH 7.4 solution, the concentration was as high as 8000 ng/mL, and there was no photosensitivity at the time of illumination, but Compound 2 exhibited very high photosensitivity when cultured at pH 6.5, IC ₅₀ A value of 95 ng/mL demonstrates a very significant tumor microcellular environment targeting.

It is known that almost all solid tumors have a slightly acidic environment, such as lung cancer, stomach cancer, esophageal cancer, breast cancer, bladder cancer, prostate cancer, pancreatic cancer, cholangiocarcinoma, rectal cancer, colon cancer, skin cancer, head and neck. Solid tumors such as cancer, ocular tumor, uterine cancer and ovarian cancer all have a slightly acidic environment, and the compound disclosed in the present patent or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same can be prepared as a photosensitizing drug for treating the above cancer.

The above is only the embodiments of the present invention, and is not intended to limit the present invention. Any modifications, equivalents, and improvements made within the spirit and scope of the present invention should be included in the scope of the present invention. Inside.

Claims (8)

1. A compound of the formula (I):

   

   Where: n = 1, 2, 3, or 4; X = O or NH; R =

   

   Or a pharmaceutically acceptable salt thereof.
2. The compound of the formula (I) according to claim 1, which is selected from the group consisting of:

   

   

   Where R=

   
3. A method of preparing a compound of the formula (I) according to claim 1, the method comprising:

   

   Where: n = 1, 2, 3, or 4; X = O or NH; R =

   

   In the first step, a compound of the formula (II) containing an acetal bond is reacted with cholesterylcarbonyl chloride in an organic solvent in an alkaline environment to obtain a compound of the formula (III);

   In the second step, a compound of the formula (IV) is reacted with a compound of the formula (III) in an organic solvent under basic conditions to obtain a compound of the formula (I);
4. The method of claim 3 wherein:

   In the first step, the solvent is selected from the group consisting of N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, chloroform and tetrahydrofuran; the reaction is carried out at a temperature of -5 to 80 ° C; The basic condition is provided by a reagent selected from the group consisting of pyridine, triethylamine, sodium hydride and 4-N,N-lutidine; the molar ratio of the compound of the formula (II) to cholesterol formyl chloride is 1:0.2-2 .

   In the second step, the solvent is selected from the group consisting of N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, chloroform and tetrahydrofuran; the reaction is carried out at a temperature of -5 to 80 ° C; The basic condition is provided by a reagent selected from the group consisting of pyridine, triethylamine, sodium hydride, and 4-N,N-lutidine; the molar ratio of the compound of the formula (IV) to the compound of the formula (III) is 1: 0.5 to 5.
5. A pharmaceutical composition comprising a therapeutically effective amount of a compound of the formula (I) according to any one of claims 1 to 2, together with a pharmaceutically acceptable carrier, diluent or excipient.
6. Use of the compound of the formula (I) according to any one of claims 1 to 2 or the pharmaceutical composition according to claim 5 for the preparation of a photodynamic drug or a photosensitizing drug.
7. Use of the compound of the formula (I) according to any one of claims 1 to 2 or the pharmaceutical composition according to claim 5 for the preparation of a medicament for treating cancer.
8. The use according to claim 7, wherein the cancer is selected from the group consisting of lung cancer, gastric cancer, esophageal cancer, breast cancer, bladder cancer, prostate cancer, pancreatic cancer, cholangiocarcinoma, rectal cancer, colon cancer, skin cancer, head and neck Cancer, eye tumors, uterine cancer and ovarian cancer, preferably breast cancer.