WO2019056376A1 - Acid-sensitive gefitinib-fluoroboronbipyrrole derivative and preparation method therefor and medical use thereof - Google Patents

Abstract

The present invention relates to a slightly acidic environment-sensitive and ketal-linked gefitinib-fluoroboronbipyrrole derivative and a preparation method therefor and a medical use thereof. The molecular structure of the derivative is as described in formula (I) of the description. In particular, the present invention relates to a gefitinib-fluoroboronbipyrrole derivative and a preparation method therefor and a pharmaceutical composition containing the compound, and the use thereof as a photosensitiser, especially the use in the treatment of cancers. In a normal tissue environment, due to the interplay between the gefitinib group and the fluoroboronbipyrroline derivative, the compound has a lower cytotoxicity. However, in the extracellular slightly-acidic environment of the tumour tissue, ketals hydrolyse and the fragments of the fluoroboronbipyrrole derivative obtained by hydrolysis exhibit an extremely high photosensitive activity, while a gefitinib derivative obtained by hydrolysis can be used as an inhibitor of epidermal growth factor receptor (EGFR) tyrosinase for inhibiting tumour growth. The compounds can be prepared into photodynamic therapeutic and chemotherapeutic double-efficacy anticancer drugs targeting both the tumour extracellular slightly-acidic environment and the EGFR tyrosine kinase.

Description

Acid-sensitive gefitinib-fluoroboron dipyrrole derivative, preparation method thereof and application in medicine Technical field

The invention belongs to the field of medicine and relates to gefitinib-fluoroboron dipyrrole derivative, a preparation method thereof and application thereof in medicine, and the invention discloses the same as a photodynamic therapy-chemotherapy dual therapeutic drug for treating cancer. use.

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. At present, photodynamic therapy has been successfully applied to 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 Treatment of ovarian cancer.

In recent years, anticancer drugs targeting the epidermal growth factor receptor (EGFR) target have received increasing attention. EGFR is a membrane receptor in a variety of malignant tumors such as glioma, breast cancer, lung cancer, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, prostate cancer, liver cancer, colon cancer, stomach cancer, etc. Overexpression in both, activation of EGFR will accelerate tumor cell proliferation, promote tumor angiogenesis, accelerate tumor metastasis, and impede tumor apoptosis.

Gefitinib is an EGFR tyrosine kinase inhibitor that is a targeted drug for the treatment of tumors. It was first marketed in Japan in 2002 and was approved as a third-line monotherapy for advanced non-small cell lung cancer in the United States and Australia in May 2003. Formally launched in China in 2005, it has been used for clinical treatment of locally advanced or metastatic non-small cell lung cancer.

The hypoxic microenvironment present in the solid tissue of the tumor results in a lower pH outside the tumor cell at that site (at 6.5 or so), while the normal tissue extracellular pH 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 present invention discloses a series of gefitinib-fluoroboron dipyrrole derivatives linked by micro-acid environmentally sensitive bonds outside the tumor tissue. In normal tissue environment, the compound has low cytotoxicity due to the interaction between the fluoroboron dipyrrole derivative group and the gefitinib group, but can be released simultaneously by hydrolysis in the extracellular micro-acid environment of the tumor tissue. Very high photosensitive active boron phthalocyanine fragment fraction and high EGFR tyrosine kinase inhibitory activity gefitinib derivative fragments. They can be prepared as a dual-targeted anti-cancer drug for tumor cell extracellular micro-acid environment and EGFR tyrosine kinase dual targeting.

Summary of the invention

The present invention relates to a microacid environment sensitive ketal linked gefitinib-fluoroboron dipyrrole derivative, a process for the preparation thereof and a pharmaceutical use. In particular, the present invention relates to a gefitinib-fluorobora dipyrrole derivative, a process for the preparation thereof, and a pharmaceutical composition containing the same, and its use as a photosensitizer, particularly in the treatment of cancer. In normal tissue environment, due to the interaction of gefitinib group and fluoroboron dipyrrole derivative, the compound has low cytotoxicity, but in the extracellular micro-acid environment of tumor tissue, the ketal is hydrolyzed and hydrolyzed. The obtained fluoroboron dipyrrole derivative fragments exhibit extremely high photosensitivity, and the hydrolyzed gefitinib derivative can be used as an epidermal growth factor receptor (EGFR) tyrosinase inhibitor to inhibit tumor growth. They can be prepared as a dual-targeted anti-cancer drug for tumor cell extracellular micro-acid environment and EGFR tyrosine kinase dual targeting.

The compound of formula (I) provided by the present invention:

Or a pharmaceutically acceptable salt thereof.

Hydrolysis reaction of compound represented by formula (I) in tumor tissue micro-acid environment (1)

The compound of formula (I) is structurally stable under normal tissue extracellular pH 7.4. The complex has low cytotoxicity due to the interaction of the fluoroboron dipyrrole derivative group and the gefitinib group.

However, in the environment where the pH outside the extracellular tissue of the tumor tissue is low (about 6.5), the ketal bond is unstable, and the compound of the formula (I) can be hydrolyzed to obtain the compound of the formula (II) and the formula (III). The structure of the compound of the formula (II) is identical and similar to that of the compounds reported in the literature ( Jiang, X.-J. et . al, Chem . Eur . J. 2016, 22, 8273-8281), and these compounds are in tumors. The cell uptake rate is high and exhibits very high photosensitivity at very low concentrations. The compound of the formula (III) is a compound very similar to the structure of gefitinib, exhibits an inhibitory effect on the epidermal growth factor receptor tyrosine kinase and inhibits tumor growth.

The present invention also provides a process for the preparation of the compound of the formula (I), the reaction equation being as follows, but not limited to the following methods:

In the first step, the organic 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 at least one agent selected from the group consisting of pyridine, triethylamine, sodium hydride and 4-N,N-lutidine; the molar ratio of the material a to p-toluenesulfonyl chloride is 1: 0.2 to 2.

In the second step, the solvent is selected from the group consisting of N,N-dimethylformamide, dimethyl sulfoxide, ethanol, and methanol; the reaction is carried out at a temperature of 30 to 120 ° C; and the basic condition is selected from the group consisting of pyridine Providing at least one of triethylamine, potassium carbonate, and sodium carbonate; the molar ratio of the intermediate a to the material b is 1:0.3-3. The material b reference method is easily synthesized from gefitinib (Tetrhedron Letters, 46(43), 7381-7384).

In the third step, the solvent is selected from the group consisting of N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, chloroform and tetrahydrofuran; the catalyst is selected from the group consisting of triethylamine, pyridine, N, N- Lutidine; the condensing agent is dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, 1 -ethyl-(3-dimethylaminopropyl)carbonyldiimide hydrochloride, O-(7-azabenzotriazol-1-yl)-bis(dimethylamino)carbonate Hexafluorophosphate, O-(benzotriazol-1-yl)-bis(dimethylamino)carbonium hexafluorophosphate, O-(benzotriazol-1-yl)-di(II) At least one of methylamino)carbonium tetrafluoroborate, O-(N-succinimidyl)-di(dimethylamino)carbonium tetrafluoroborate; and/or The condensation activator is selected from the group consisting of 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, N-hydroxysuccinimide, and N-hydroxyphthalimide; The reaction is carried out at a temperature of -5 to 80 ° C; the molar ratio of the compound of the formula (IV) to the intermediate c is 1:0.4 to 4; the molar ratio of the compound of the formula (IV) to the condensing agent is 1: 0.4 to 4. The compound of the formula (IV) is synthesized by a reference method ( Jiang, X.-J. et . al , Chem . Eur . J. 2016 , 22 , 8273-8281 ).

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 invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of 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 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 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, uterine and ovarian 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.

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 using a suitable 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 500 to 900 nm, preferably 550 to 750 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 using a binder that melts at rectal temperatures, for example, cocoa butter or poly Prepared with ethylene glycol.

An aqueous suspension, an isotonic physiological saline solution or a sterile and injectable solution comprising a pharmacologically compatible dispersant and/or wetting agent for parenteral, intranasal or intraocular administration Apply. 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.

"TsCl" is p-toluenesulfonyl chloride.

Detailed ways

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 material b reference method is easily synthesized from gefitinib (Tetrhedron Letters, 46(43), 7381-7384). The compound of the formula (IV) is synthesized by a reference method ( Jiang, X.-J. et . al , Chem . Eur . J. 2016 , 22 , 8273-8281 ).

Nuclear Magnetic Resonance Instrument: Bruker ARX-400 high resolution high resolution nuclear magnetic resonance instrument.

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

MTT test instrument: 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).

Example 1: Synthesis of a compound of formula (I)

step 1

In an ice water bath, material a (1.64 g, 10 mmol), p-toluenesulfonyl chloride (1.9 g, 10 mmol) and pyridine (1.6 g, 20.2 mmol) were added to dichloromethane (45 mL) and the mixture was warmed to room temperature and stirring was continued. After the reaction was continued for 15 hours, the reaction was stopped, and water (300 mL) was added to the reaction mixture, and the mixture was stirred and left to dryness. The organic phase was collected, dried over anhydrous sodium sulfate and evaporated under reduced pressure. The solvent was methylene chloride/methanol (10:1). MS (ESI): m/z = 319 [M+H] ⁺ .

Step 2

Intermediate a (1.59 g, 5 mmol), material b (2.16 g, 5 mmol) was added to N,N-dimethylformamide (50 mL), and anhydrous potassium carbonate (1.38 g) was added to the reaction mixture. 10 mmol), the reaction solution was heated to 90 ° C, and the reaction was stopped after stirring for 15 hours. After the reaction was stopped, the organic solvent was removed under reduced pressure. The crude product was added to dichloromethane (100 mL), and then stirred and dissolved. Then, 200 mL of water was added and stirred. After standing, the organic phase was collected and dried over anhydrous sodium sulfate. Purified, the eluent was methylene chloride / methanol (15:1) toield ^{1 H NMR (400MHz, DMSO-} d 6): δ9.50 (s, 1H, NH), 8.50 (s, 1H, Ar-H), 8.08-8.15 (m, 1H, Ar-H), 7.70-7.83 (m, 2H, Ar-H), 7.38-7.49 (m, 1H, Ar-H), 7.20 (s, 1H, Ar-H), 4.10-4.26 (m, 4H, CH ₂ ), 3.55-3.72 ( m, 10H, CH ₂ ), 2.44 - 2.55 (m, 2H, CH ₂ ), 2.34 - 2.45 (m, 4H, CH ₂ ), 1.94 - 2.05 (m, 2H, CH ₂ ), 1.41 (s, 6H, CH ₃ ). MS (ESI): m/z = 579 [M] ⁺ .

Step 3

The compound of the formula (IV) (1.06 g, 1.0 mmol) and the intermediate c (0.64 g, 2 mmol) were dissolved in N,N-dimethylformamide (30 mL), and the temperature of the solution was dropped to 0 ° C and stirred for 0.5 hour. Add 1-ethyl-(3-dimethylaminopropyl)carbonyldiimide hydrochloride (EDC.HCL) (767 mg, 4 mmol), 4-N,N-dimethylpyridine (244 mg, 2 mmol) The temperature of the reaction liquid was raised to react at 60 ° C for 24 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was evaporated. The solvent was evaporated to dryness, and the residue was purified by silica gel chromatography. The eluent was chloroform/methanol (9:1 v/v). Drying under reduced pressure gave a brown solid (yield: Compound: _{^{1 H NMR (400MHz, CDCl 3}} ): δ8.50 (s, 1H, Ar-H), 8.06-8.15 (m, 3H, Ar-H, CH = CH), 7.70-7.83 (m, 2H, Ar- H), 7.38-7.49 (m, 1H, Ar-H), 7.57 (d, J = 8.4 Hz, 4H, ArH), 7.55 (d, J = 16.4 Hz, 2H, CH = CH), 7.20 (s, 1H, Ar-H), 7.13 (d, J = 8.4 Hz, 2H, ArH), 7.05 (d, J = 8.4 Hz, 2H, ArH), 6.98 (d, J = 8.4 Hz, 4H, ArH), 4.70 -4.76 (m, 4H, CH ₂ ), 4.10-4.26 (m, 14H, CH ₂ ), 3.55-3.78 (m, 24H, CH ₂ ), 3.42-3.51 (m, 14H, CH ₂ ), 3.24 (s , 3H, CH ₃ ), 2.34 - 2.55 (m, 10H, CH ₂ ), 1.94 - 2.05 (m, 4H, CH ₂ ), 1.47 (s, 3H, CH ₃ ), 1.41 (s, 12H, CH ₃ ) . MS (ESI): m / z = 2183 [M + H] +.

Test Example 1: Hydrolysis of a compound of the formula (I)

The compound of formula (I) (218 mg, 0.1 mmol) was dissolved in 50 mL of tetrahydrofuran and 50 mL of a pH 6.5 PBS solution was added. The pH of the solution was maintained at 6.5 and the reaction was stirred at room temperature for 24 hours. The organic solvent was distilled off under reduced pressure, dichloromethane (200 mL) was added, and the mixture was shaken, and the organic phase was collected. The organic phase is dried over anhydrous sodium sulfate and evaporated under reduced pressure. The crude product is purified eluting with silica gel column eluting with methylene chloride/methanol (20:1) to give red solid (II) compound (34 mg, 30 %) and a brown solid compound of formula (III) (18 mg, 38%).

Compounds of formula (II): ¹ H NMR (400 MHz, DMSO-d ₆ )): δ 8.06 (d, J = 16.4 Hz, 2H, CH=CH), 7.57 (d, J = 8.4 Hz, 4H, ArH) , 7.45 (d, J = 16.4 Hz, 2H, CH = CH), 7.33 (d, J = 8.4 Hz, 2H, ArH), 7.15 (d, J = 8.4 Hz, 2H, ArH), 7.02 (d, J) = 8.4 Hz, 4H, ArH), 4.70-4.76 (m, 4H, CH ₂ ), 4.16-4.20 (m, 2H, CH ₂ ), 3.76-3.78 (m, 2H, CH ₂ ), 3.70-3.74 (m , 2H, CH ₂ ), 3.66-3.68 (m, 2H, CH ₂ ), 3.55-3.59 (m, 2H, CH ₂ ), 3.42-3.45 (m, 2H, CH ₂ ), 3.24 (s, 3H, CH ₃ ), 1.47 (s, 3H, CH ₃ ); MS (ESI): m/z = 1150 [M] ⁺ .

Compounds of formula (III): ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 9.50 (s, 1H, NH), 8.50 (s, 1H, Ar-H), 8.07-8.14 (m, 1H, Ar- H), 7.70-7.85 (m, 2H, Ar-H), 7.37-7.49 (m, 1H, Ar-H), 7.20 (s, 1H, Ar-H), 3.96-4.26 (m, 6H, CH ₂ ), 3.55-3.72 (m, 4H, CH 2), 2.94-2.55 (m, 2H, CH 2), 2.34-2.45 (m, 4H, CH 2), 1.94-2.05 (m, 2H, CH 2); MS (ESI): m/z = 437 [M+H] ⁺ .

Test Example 2: In vitro anti-tumor cell photosensitivity test

Test sample: a compound of the formula (I), a compound of the formula (II) and a compound of the formula (III)

Test cells: human lung cancer cell PC9

Main reagents: 1) DMEM complete medium: Add 500,000 U of penicillin/streptomycin, 56 mL of fetal bovine serum to 500 mL DMEM 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:

The test sample was formulated into a mother liquor at a concentration of 1 mM in DMSO; 100 μL of 1 mg/mL mother liquor was taken during the experiment, and 1.15 mL of 0.5% (w/w) polyoxyethylene castor oil pH 7.4 PBS and pH 6.5 PBS buffer were added to prepare the test solution. The solution was made into 80 μg/mL, and diluted with different PBS buffers into different concentrations of the drug solution. The pH value of the drug solution was kept unchanged during the dilution process. After the drug solution was prepared, it was allowed to stand at room temperature for 24 hours, and then subjected to cell dosing culture. The final concentration of DMSO in each drug and negative control group was < 1%. Adherent tumor cells in logarithmic growth phase were selected and digested with trypsin, and mixed with DMEM medium containing 10% fetal bovine serum to prepare a suitable concentration of cell suspension, inoculated in 96-well culture plate at 37 ° C, 5% Incubate for 24 hours under CO ₂ conditions. Then, 100 μL of each test drug, solvent and culture solution of different concentrations were added, respectively, and 3 parallel holes per group. After mixing, they were divided into two groups: light and dark. After co-cultivation for 2 hours, the medium was discarded, and the medium containing no test or control substance was added again at 37 ° C under 5% CO ₂ . Continue to train for 24 hours. After 24 hours, add 5 mg/mL MTT per well, 20 μL, incubate for 4 hours at 37 ° C, 5% CO ₂ , discard the supernatant, add 200 μL of DMSO to each well, shake for 10 minutes, and measure the absorbance by microplate reader. The measurement wavelength was 570 nm and the excitation 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 .

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

Experimental results:

Table 1 IC ₅₀ (nM) values of human lung cancer cell PC9 when the drug is illuminated

drug	Cell pH 7.4 liquid culture	Cell pH 6.5 liquid culture
Compound of formula (I)	>5000	45
Compound of formula (II)	88	95
Compound of formula (III)	85	78

The experimental results showed that all the tested compounds showed no cytotoxicity at concentrations up to 5000 nM in the dark, but in the case of illumination, the compound of formula (II) and the compound of formula (III) were applied to human lung cancer at pH 6.5 and pH 7.4. PC9 cells the lethal concentration IC ₅₀ values between 78-95nM. However, when the compound of the formula (I) is cultured in a liquid solution of pH 7.4, the concentration is as high as 5000 nM, and there is no photosensitivity at the time of illumination, but the compound of the formula (I) exhibits a very high photosensitivity when the cells are cultured at pH 6.5. activity, IC ₅₀ value of 45nM, this is due to the formula (I) in a tumor acidic environment obtained by hydrolyzing highly cytotoxic compound of formula (II) and a compound of formula (III).

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 (5)

1. A compound of formula (I):

   

   Or a pharmaceutically acceptable salt thereof.
2. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1 and a pharmaceutically acceptable carrier.
3. Use of a compound according to claim 1 or a pharmaceutical composition according to claim 2 for the preparation of a photodynamic drug or a photosensitizing drug.
4. Use of a compound according to claim 1 or a pharmaceutical composition according to claim 2 for the preparation of a medicament for the treatment of cancer.
5. The use according to claim 4, 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.