TWI635860B - Use of acridine derivatives for preparing pharmaceutical compositions for treating urothelial cancer - Google Patents

Abstract

Use of an acridine derivative for the preparation of a pharmaceutical composition for the treatment of urinary tract epithelial cancer, the pharmaceutical composition comprising an effective amount of an acridine derivative.

Description

Use of acridine derivatives for the preparation of a pharmaceutical composition for treating urinary tract epithelial cancer

The present invention relates to the use of an acridine derivative for the preparation of a pharmaceutical composition for the treatment of urinary tract epithelial cancer.

Urothal epithelial (urothelium) refers to the epithelial tissue distributed in the urine transport and storage organs of the urinary system, and is also called histologically called transitional epithelium. Urothelial cancer is derived from the urinary tract epithelium of the lesion, so it is also called transitional cell carcinoma (TCC). Urinary tract epithelial cancer is a malignant tumor that occurs in all urinary tract epithelial cells, including bladder cancer, urethral cancer, ureteral cancer, and renal pelvic cancer. Among them, bladder cancer accounts for about 60%, and the recurrence rate is quite high.

Diagnosis of bladder cancer In addition to general urine tests to determine the cause of hematuria, it also includes urine cytology, ultrasound, transurethral ultrasonography (TUS), computed tomography (CT), and magnetic resonance imaging ( MRI), or a further invasive examination, wherein the invasive examination includes cystoscopy, intravenous or renal pelvic radiography or cystoscopy combined with biopsy. Cystoscopy is to enter the bladder through the urethra through the urethra, and to observe the lesions in the bladder by endoscope; X-ray photography is to insert the developer intravenously, or insert the ureter catheter into the ureter or ureter through the catheter. The renal pelvis, ureter and bladder are developed to check the urinary tract system.

In general, bladder cancer is treated with transurethral resection of bladder tumor (TURBT), and a bladder endoscope is inserted through the urethra to remove the bladder tumor tissue. After transurethral resection of the bladder tumor, bladder drug perfusion therapy is generally used. The chemotherapeutic drug or bacillus calmette-guerin (BCG) is injected into the bladder through the urethra through the urethra and left for 60 minutes to kill. Cancer cells that have not been removed or are not visible to the naked eye, in order to improve the treatment rate and reduce the chance of recurrence.

At present, in the treatment of urinary tract epithelial cancer, no method for treating or adjuvant treatment of urinary tract epithelial cancer after diagnosis and the use of the pharmaceutical composition thereof have been found. Therefore, how to provide a method for immediate treatment or adjuvant treatment of urinary tract epithelial cancer after diagnosis and use of the pharmaceutical composition thereof is an extremely difficult task in the industry.

In view of the above problems, it is an object of the present invention to provide a pharmaceutical composition for the preparation of a pharmaceutical composition which can assist or immediately treat urinary tract epithelial cancer after diagnosis.

To achieve the above object, the use of an acridine derivative according to the present invention for the preparation of a pharmaceutical composition for the treatment of urinary tract epithelial cancer, wherein the pharmaceutical composition comprises an effective amount of an acridine derivative.

In one embodiment, the pharmaceutical composition is formulated to illuminate with blue light.

In one embodiment, the acridine derivative is acridine orange and its salts, acridine yellow and its salts, acridine red and its salts, 9-amino-6-chloro-2-methoxyindole Pyridine and its salts, alkylated derivatives, methylated derivatives, polymer derivatives, or combinations thereof.

In one embodiment, a method of making a pharmaceutical composition comprises preparing a perfusate comprising an effective amount of an acridine derivative and an acidic buffer.

In one embodiment, the acridine derivative concentration in the perfusate is greater than 1.0 [mu]g/mL.

In one embodiment, the pH of the perfusate is between pH 5 and pH 8.

In one embodiment, the acidic buffer is an acetate buffer, a monophosphate buffer, a citrate buffer, a tris buffer, or a combination thereof.

In one embodiment, the pharmaceutical composition is required to infuse perfusate into at least a portion of the urinary system during treatment.

In one embodiment, the acridine derivative is in the urinary system at a concentration between 1.0 [mu]g/mL and 5.0 mg/mL.

In one embodiment, the perfusate is perfused to at least a portion of the urinary system by perfusion through a catheter.

In one embodiment, the catheter is a catheter of an endoscope group and the blue light is emitted by the endoscope assembly.

In one embodiment, the combination of blue light illumination during treatment refers to at least a portion of the urinary system being illuminated by an external light source.

In one embodiment, the peak wavelength of blue light is between 400 nm and 490 nm.

In one embodiment, the time to illuminate the blue light is greater than 25 seconds.

In one embodiment, the time to illuminate the blue light is between 25 seconds and 900 seconds.

In summary, the present invention provides an acridine derivative for the preparation of a pharmaceutical composition for treating urinary tract epithelial cancer, when the acridine derivative is used for the detection of urinary epithelial cancer cells, if the medical staff is inspecting When the diseased cells/cancer cells are found, photodynamic therapy can be performed immediately, and the affected part is irradiated with blue light to instantly kill or reduce the number of urinary tract epithelial cancer cells during the detection of urinary tract epithelial cancer cells. In this way, the treatment can be performed at the first time when the cancer cells are detected, thereby simplifying the complicated examination and treatment process of the patient.

Hereinafter, the use of an acridine derivative for the preparation of a pharmaceutical composition for treating urinary tract epithelial cancer according to a preferred embodiment of the present invention will be described with reference to the accompanying drawings, wherein the same elements will be described with the same reference numerals.

In order to make this article more obvious and easy to understand, the terms used in this article are specifically defined and described below:

The term "treatment" encompasses the provision or administration of any treatment for a disease of a mammal, including a human. The so-called treatment and the inclusion of inhibition of disease, prevention of disease development, slowing down the disease; or stimulating inefficient physiological processes. The above nouns include obtaining an ideal pharmacological and/or physiological effect covering the treatment of any pathological condition or condition in a mammal, including humans. The above effects may be prophylactic, i.e., capable of completely or partially preventing the condition and or associated symptoms; or may be therapeutic, i.e., capable of partially or completely curing the condition and/or the adverse effects caused by the condition. Accordingly, the present invention provides for the treatment and prevention. It contains (1) preventing or recurring a patient who may have a disease but has not yet developed symptoms; (2) inhibiting the disease, such as preventing the development of the disease; (3) stopping or terminating the disease or at least the associated symptoms, leaving the patient no longer Suffering from the disease or its symptoms; or stimulating inefficient physiological processes; and (4) mitigating, mitigating or ameliorating the condition or related symptoms, wherein the term "improvement" broadly refers to the ability to reduce the intensity of at least one parameter, such as inflammation. , pain, and/or tumor size.

"Individual" or "patient" means a mammal, preferably a human. Mammals include farm animals, sport animals, pets, primates, horses, dogs, cats, mice, and hamsters.

"Composition" means a mixture which usually contains a carrier, such as a pharmaceutically acceptable carrier or a conventional excipient, and is suitable for administration to a patient for treatment, diagnosis or prophylaxis. It may comprise a cell culture in which a multi-peptide or polynucleotide is present in the cell or medium. For example, the oral compositions can be solutions, suspensions, lozenges, pills, capsules, sustained release formulations, mouthwash or powder.

"Pharmaceutically acceptable carrier" means a non-toxic solid, semi-solid or liquid, filler, diluent, encapsulating material, formulation adjuvant, or any of the conventional types of excipients. The pharmaceutically acceptable carrier is non-toxic to the patient at the dosages and concentrations employed and is compatible with the other ingredients of the formulation. Such carriers include, but are not limited to, saline, buffer solutions, dextrose, water, glycerol, ethanol, and combinations thereof.

By "effective dose" is meant that a desired amount of effect can be achieved in a patient when administered to a patient. For example, an effective amount of a pharmaceutical composition comprising an acridine derivative of the present invention administered to a patient is an amount required to prevent and/or treat urinary tract epithelial cancer. The actual amount of use will vary with the purpose of the treatment, and one skilled in the art can determine this amount using known techniques. It is also known in the related art that doses may need to be adjusted for systemic or topical administration, age, weight, general health, sex, eating habits, time of administration, drug interaction, and severity, and are known in the art. The quantity can be determined by routine experimentation.

"Acridine derivative" generally refers to an acridine dye which can be used as a fluorescent dye, and is fat-soluble and slightly alkaline. For example, the acridine dye as a fluorescent dye may be acridine orange and its salts, acridine yellow and its salts, acridine red and salts thereof, Or 9-amino-6-chloro-2-methoxyacridine (ACMA) and its salts, alkylated derivatives, methylated derivatives, polymers Derivative, or a combination of the above.

Acridine orange, chemical formula C ₁₇ H ₁₉ N ₃ , chemical name 3,6-dimethylamino acridine (N, N, N', N'-Tetramethylacridine-3, 6-diamine), with inorganic acids Salts are formed (for example, chloride salts, zinc chloride salts). Acridine orange is a kind of nucleic acid dye, and it is soluble in water and alcohol. Its absorption and emission spectra will vary according to its concentration. The excitation wavelength of acridine orange is roughly between 440nm and 505nm. The emission wavelength is between 520 nm and 650 nm. For example, when combined with DNA, the excitation wavelength is 502 nm and the emission wavelength is 525 nm (green); when combined with RNA, the excitation wavelength is 460 nm (blue) and the emission wavelength is 650 nm (red). The chemical structure of acridine orange is as follows:

Acridine yellow, chemical formula C ₁₅ H ₁₅ N ₃ Cl, chemical name 3,6-diamino-2,7-dimethylacridine (3,6-diamine-2,7-dimethylacridine) is a kind of coal from coal The dye extracted from tar has a brownish red appearance. Acridine yellow is soluble in alcohol, and its excitation wavelength is 420nm~470nm, and the emission wavelength is 525nm~550nm. The chemical structure of acridine yellow is as follows:

Acridine red, the chemical formula is C ₁₅ H ₁₅ ClN ₂ O, the chemical name is methyl-[6-(methylamino) xanthen-3-ylidene] azanium chloride, and its excitation wavelength is 455 nm. ~600nm, the emission wavelength is 560nm~680nm. The chemical structure of acridine red is as follows:

Acridine, a chemical formula of C ₁₄ H ₁₄ ClN ₃ , chemically known as 3,6-diamino-10-methylacridinium chloride, which is excited The wavelength was 452 nm and the emission wavelength was 510 nm. The chemical structural formula of methyl acridine is as follows:

9-amino-6-chloro-2-methoxyacridine (ACMA), chemical formula C ₁₄ H ₁₁ ClN ₂ O, excitation wavelength 412 nm ~ 430 nm, luminescence The wavelength is 471 nm to 474 nm. The chemical structure of ACMA is as follows:

Quiancrine, chemical formula C ₂₃ H ₃₀ ClN ₃ O, chemical name N4-(6-chloro-2-methoxy-9-acridinyl)-N1,N1-diethyl-1 , 4-pentanediamine ((RS)-N'-(6-Chloro-2-methoxy-acridin-9-yl)-N,N-diethylpentane-1,4-diamine), which has an excitation wavelength of 445 nm and emits light. The wavelength is 500 nm. The chemical structure of quinacrine is as follows:

Autophagy is a cellular response triggered by cellular stress (such as nutrient deficiencies). Autophagy produces preautophagosomes through reorganization of intracellular membrane structures and a series of messages. An isolation membrane/phagophore is formed, and after the bilayer membrane structure coats the cytoplasm to become an autophagosome, the autophagosome binds to lysosomes as autophagosomes. (autophagolysosomes), which decomposes its coated contents. When the lysosome is fused with the autophagosome, the organelle or protein encapsulated by the autophagosome can be digested by the enzyme in the lysosome, and the pH in the acid bubble is acidic, if it is stained by the acridine derivative. Small particles of fluorescent color are present, which are acidic vesicular organelles (AVOs). The accumulation of acridine derivatives in acidic organelles is related to the pH value, which can be combined with acidic cystic organelles, and is often used to detect the presence of acidic cystic organelles, thereby determining cells. Whether it has died. When cells are stained and stimulated by blue light, the cytoplasm and nucleoli will appear bright green or orange-red, while lysosomes or autophagic lysosomes will be bright red or orange-red (Reference Kanzawa, T., Kondo, Y , Ito, H. et al.: Induction of autophagic cell death in malignant glioma cells by arsenic trioxide. Cancer Res, 63: 2103, 2003. Lin, JF, Lin, YC, Lin, YH et al.: Zoledronic acid induces Autophagic cell death in human prostate cancer cells. J Urol, 185: 1490, 2011.).

Photosensitizer, in photochemical reactions, a class of molecules that only absorb photons and transfer energy to molecules that cannot absorb photons, causing them to undergo chemical reactions, while not participating in chemical reactions themselves, and returning to the original The state of this type of molecule is called a photosensitizer. The photochemical reaction initiated by the photosensitizer is called a photo-sensitive reaction. Photosensitive reactions involving the biological effects of aerobic molecules are usually called photodynamic reactions, and drugs that cause photodynamic reactions to destroy cell structures are called photodynamic drugs, that is, photosensitizing drugs.

Photodynamic therapy (PDT), formerly known as photoradiation therapy (PRT) or photochemical therapy (PCT), is a new technology for photodiagnosis and treatment of disease diagnosis. The basic process of photodynamic therapy is that when the endogenous or exogenous photosensitive substance in the biological tissue is illuminated by the corresponding wavelength (visible light, near-infrared light or ultraviolet light), the photon energy is absorbed, and the ground state becomes an excited state. Since the photosensitive material in the excited state is unstable, it will rapidly release energy and return to the ground state through physical de-excitation or chemical de-excitation, and the physical de-excitation process can generate fluorescence, and the disease can be diagnosed by analyzing the fluorescence spectrum; The chemical de-excitation process can generate a large amount of active oxygen, the most important of which is singlet oxygen, which can interact with various biological macromolecules, damage its cell structure or affect cell function, thus producing therapeutic effects. For example, killing tumor cells/cancer cells.

The present invention provides a use of an acridine derivative for the preparation of a pharmaceutical composition for treating urinary tract epithelial cancer, which is a pharmaceutical composition for the preparation of a medicament for treating or preventing urinary tract epithelial cancer. Herein, the pharmaceutical composition can be used for immediate diagnosis of urinary tract epithelial cancer, or combined with bladder drug perfusion therapy for adjuvant therapy.

A pharmaceutical composition for treating or preventing urinary tract epithelial cancer is provided to a desired individual for treatment, and is combined with blue light irradiation. Among them, the acridine derivative can be a photosensitizer, and the acid vesicles (AVOs) formed by the cancer cells during autophagy can be observed by staining with acridine derivatives, and can be used as an examination for cancer.

A pharmaceutical composition for treating or preventing urinary tract epithelial cancer can be administered by a known drug delivery method such as intravenous injection, smearing or infusion.

In one embodiment, the method for preparing the above pharmaceutical composition comprises: preparing a perfusate comprising an effective amount of an acridine derivative and an acidic buffer, wherein the acidic buffer is an acetate buffer, monophosphate The salt buffer, the citrate buffer, the Tris (hydroxymethyl) aminomethane, Tris buffer or a combination thereof, so that the pH of the perfusate is between pH 5 and pH 8. Wherein, the concentration of the acridine derivative in the perfusate is greater than 1.0 μg/mL, and the concentration of the acridine derivative is preferably between 1.0 μg/mL and 5.0 mg/mL.

The pharmaceutical composition can be treated after the patient has confirmed that it is a urinary tract epithelial cancer, and the means of diagnosis can be a conventional technique, and thus it is no longer repeated. At the time of treatment, the pharmaceutical composition can be carried out by bladder drug perfusion therapy, or even directly mixed into a perfusate containing a bladder drug (for example, a chemotherapeutic drug, BCG, etc.) for adjuvant treatment, that is, the pharmaceutical composition may further comprise a chemotherapeutic drug. , BCG or other urinary tract epithelial cancer treatment drugs. The following examples are illustrated by the use of a pharmaceutical composition to assist in the diagnosis of urinary tract epithelial cancer, followed by treatment with blue light, to achieve immediate treatment of urinary tract epithelial cancer.

Referring to Figure 1, in the urinary tract test, a perfusion solution containing an effective dose can be infused into the affected part by the catheter, that is, at least part of the urinary system (kidney K, bladder B, ureter Ur, and urethra Ua). To perform staining of cancer cells. It should be noted that in addition to the acridine derivative, there may be other dyes in the perfusate that can be used for cancer cell detection.

For example, for bladder B perfusion, catheter C can be a separate tube or set in a group with a bladder endoscope. Usually, during bladder infusion, the patient is lying flat or in a lithotomy position, and a thin flexible catheter is inserted into the bladder from the urethra. The urine can be led through the hole O of the catheter, and the pharmaceutical composition is then About 30 to 50 ml) is infused into the bladder.

In the detection process, the acridine derivative is firstly used as a fluorescent dyeing property, and a perfusion solution which has been contacted by the urinary system is first taken out by a catheter to observe, or directly observed by an endoscope to assist in confirming whether or not There are lesions or cancerous tumor cells. Taking acridine orange as an example, when an AVO of orange-red color is found during the examination, it indicates that the patient has a diseased or cancerous tumor cell, and the medical staff immediately starts blue light irradiation for photodynamic therapy.

The purpose of blue light irradiation is to allow the photosensitive material or acridine derivative to generate an excited state to emit singlet oxygen. When the excitation peak wavelength required for the complex acridine derivative is not blue light, it may be replaced by other radiation light corresponding to the excitation peak wavelength of the photosensitive material or the acridine derivative, such as red light, green light, violet light or ultraviolet light.

The timing of blue light exposure to an acridine derivative (or medical composition) may be when the perfusate or medical composition has entered the urinary system, or before the perfusate or medical composition is administered to the patient. If the perfusate or medical composition is administered to the patient prior to administration, the perfusate or medical composition can be directly administered with blue light for administration. Here, the infusion solution or the medical composition has entered the urinary system, and then blue light irradiation is taken as an example. Wherein, the blue light source can be provided by an LED light bulb, and the blue light can be indirectly irradiated to the perfusate or the medical composition through the skin of the human body by an external blue light source or a combination of other light sources and a filter to generate blue light. Here, the peak wavelength of the blue light is between 400 nm and 490 nm, and when the corresponding pharmaceutical composition is acridine orange, it is preferably between 430 nm and 450 nm; the irradiation time of the blue light is greater than 25 seconds. Preferably, the system is between 25 seconds and 900 seconds; and the blue light illumination distance is between 0.1 cm and 15 cm.

It should be noted that the blue light can also be emitted by the light source carried or set by the endoscope group, so that the endoscope group protrudes into the human body, for example, after extending into the bladder along the urethra, and is equipped with the endoscope group. The blue light source above can be directly irradiated to a perfusate or medical composition containing an acridine derivative. Wherein the concentration of the acridine derivative in the urinary system is between 1.0 μg/mL and 5.0 mg/mL. In addition, the endoscope group can also be equipped with a catheter to assist in the procedure of perfusion. In order to allow the drug to fully contact the mucous membrane of the bladder, the patient is usually allowed to rest in different postures for at least 60 minutes, preferably 1 to 3 hours, but not limited thereto. After the break or when the urine rises, the patient discharges the urine by himself. liquid.

Therefore, when performing cystoscopy or other urinary system examination, the perfusion solution containing acridine derivative can be perfused, and the acridine derivative is used as a fluorescent dye to help confirm the presence of lesions or cancerous. Tumor cells. When AVOs stained or fluorescently labeled are found during the examination, it indicates that the patient has a diseased or cancerous tumor cell, and the medical staff can perform blue light irradiation for a short time for photodynamic therapy. This system utilizes the acridine derivative as a photosensitizer to produce singlet oxygen after exposure to blue light, and to kill or reduce cancerous or cancerous tumor cells.

Men's bladder cancer cell lines have been mentioned in the references (Lin, YC, Lin, JF, Wen, SI et al.: Inhibition of High Basal Level of Autophagy Induces Apoptosis in Human Bladder Cancer Cells. J Urol, 2015). In the presence of highly activated autophagy, autophagy is required for cancer cell growth; and by promoting apoptotic cell death, it can inhibit autophagy before cancer cells (pro-survival). Autophagy), which in turn reduces the survival rate of urinary tract epithelial cancer cells. Some photosensitizers have been reported to have the effect of inducing apoptosis.

The invention also provides a method for treating urinary tract epithelial cancer, wherein the method comprises the steps of: preparing a perfusate comprising an effective amount of an acridine derivative; and illuminating the perfusate with blue light.

Hereinafter, it will be confirmed by a plurality of experimental examples that the pharmaceutical composition having an acridine derivative can have an effect of treating urothelial cancer cells after being irradiated with blue light.

Materials and analytical methods

Chemicals and cell lines: All chemicals are prepared as stock solutions using standard procedures. Human immortalized urothelial cell line (SV-Huc-1) and bladder cancer cell lines (RT4, 5637 and T24) were purchased from the Bioresource Collection and Research Center. BCRC).

Blue light source: In the experimental example, six blue LED bulbs (peak wavelength 443.7 nm) were used, soldered to a circuit board and placed in an aluminum foil-wrapped carton, and the LED bulb was driven by a power converter. The carton covers a 96-well cell culture dish or a 10 cm culture dish.

Fluorescence imaging analysis of cells using a Nikon inverted microscope Eclipse Ti-E equipped with a 130 W fluorescent source and a band-pass excitation filter from 465 nm to 495 nm And a 590 nm long-pass barrier filter.

Cell viability assays: In this study, reference is made to the literature (Lin, JF, Tsai, TF, Liao, PC et al.: Benzyl isothiocyanate induces protective autophagy in human prostate cancer cells via inhibition of mTOR signaling. Carcinogenesis, 34: 406, 2013.) The activity of the cells was determined by WST-1 reagent (Roche Diagnostics, Mannheim, Germany), and the analysis process avoided illumination.

Statistical analysis: Data were expressed as mean ± standard deviation and calculated by Student t-test for statistical significance, and a P value of less than 0.05 was considered statistically significant.

The first experimental example: the effect of acridine derivatives or blue light on the activity of bladder cancer cell line 5637 and bladder cancer cell line T24

As shown in Fig. 2A to Fig. 2D, in order to study whether the acridine derivative or blue light irradiation has an effect on the cell activity of bladder cancer cells, the bladder cancer cell line 5637 and T24 are first cultured in a 96-well cell culture plate for 24 hours. The following different treatments are carried out separately, wherein the acridine derivative is exemplified by acridine orange (AO): (A) the culture solution is replaced with 0-2.5 μg/mL AO for 30 minutes, but not blue light; (B The blue light is irradiated for different time from 0 to 300 seconds, but the culture solution is not added with AO; (C) the culture solution is replaced with 1 μg/mL AO for 30 minutes, and the blue light is irradiated for 0 to 300 seconds; (D) the culture solution is After replacing it with 0~2 μg/mL AO for 30 minutes, the blue light was irradiated for 30 seconds. This experiment avoided illumination in the AO treatment process.

As a result, as shown in Fig. 2A, bladder cancer cell lines 5637 and T24 were not exposed to blue light, and the cell viability of 5637 and T24 was not different from that of the control group (AO concentration of 0) only after treatment with different AO concentrations for 30 minutes. As shown in Fig. 2B, the bladder cancer cell lines 5637 and T24 without acridine orange were irradiated with blue light for different periods of time, and the cell viability of bladder cancer cell lines 5637 and T24 was not different from that of the control group (no blue light). Therefore, bladder cancer cell lines 5637 and T24 have only AO-treated or blue-light-only groups, and have no effect on cell viability. As shown in Fig. 2C, bladder cancer cell lines 5637 and T24 treated with AO (AO concentration of 1 μg/mL) were exposed to blue light for 30 seconds, and the activity of 5637 and T24 cells was significantly higher than that of the control group. Reduced. As shown in Fig. 2D, after 30 seconds of blue light irradiation, when the AO concentration was greater than or equal to 1 μg/mL, the cell viability of bladder cancer cell lines 5637 and T24 was significantly reduced compared with the control group, and the higher the AO concentration, 5637 and The lower the activity of T24 cells.

In addition, based on the results of the first experimental example, the concentration of the subsequent AO treatment was selected to be 1 μg/mL, and the blue light irradiation time was 30 seconds. Thus, the combination of acridine derivative treatment and blue light irradiation may be referred to as acridine derivative. Photodynamic therapy (AO-PDT).

Second Example: Effect of acridine derivatives and blue light on normal cells (human immortal urothelial cell line SV-Huc-1)

As shown in FIG. 3A to FIG. 3C, since the bladder cancer cells are subjected to AO treatment and blue light irradiation, the cell activity is significantly reduced. In order to understand whether the treatment of AO and blue light irradiation has an effect on cancer cells, this experiment (Fig. 3A and FIG. 3B) The bladder cancer cell line 5637 and T24 were cultured in a 96-well cell culture plate for 24 hours, and then the culture solution was replaced with 1 μg/mL AO for 30 minutes, and then the blue light was irradiated for 30 seconds to complete the blue light. After the irradiation, the culture solution (normal culture medium containing 10% serum) was replaced, and cell viability was calculated after 1 hour, 4 hours, and 24 hours of culture (this was the treatment group, and the control group was not subjected to AO treatment or performed). Blue light illumination). 3C shows the treatment conditions of the human immortal urothelial cells SV-Huc-1 and the above treatment group, and compared with the results of the same treatment of bladder cancer cells T24 24 hours after the blue light is irradiated (this is In the treatment group, the control group was not subjected to AO treatment or blue light irradiation).

As can be seen from Fig. 3A and Fig. 3B, the cell viability of the bladder cancer cell lines 5637 and T24 which were photodynamically treated with acridine derivatives was significantly reduced as the culture time increased. As shown in Fig. 3C, even if the human immortal urothelial cells SV-Huc-1 was cultured for 24 hours, the cell activity did not decrease, indicating that AO and blue light had no effect on normal cell activity.

Third Example: Effect of acridine derivatives and blue light on normal cells (human immortal urothelial cell line SV-Huc-1)

Please refer to FIG. 4A to FIG. 5D simultaneously, and culture the human immortal urothelial cell line SV-Huc-1, bladder cancer cells RT4, 5637 and T24 into a 96-well cell culture dish for 24 hours, and replace the culture solution with 1 μg. After treatment with /mL AO for 30 minutes, the blue light was irradiated for 30 seconds (this is the treatment group, the control group is not subjected to AO treatment or blue light irradiation); after the blue light irradiation, the culture medium (normal medium containing 10% serum) is replaced, and then The plates were placed in a Cytation 5 Imaging Reader and the cells were automatically counted by the detector's built-in software (Biotek) at 0, 6, 12, 18 and 24 hours. The cell confluence was measured with a cell fusion degree of 0 hours (1.00), and the results are shown in FIGS. 4A to 4D, and the cell morphology recorded by the automatic photographing is as shown in FIGS. 5A to 5D. It can be seen from the results that AO treatment and blue light irradiation have no effect on the activity of SV-Huc-1 cells, but the cell viability of bladder cancer cells RT4, 5637 and T24 after AO treatment and blue light irradiation is significantly reduced, showing that AO and blue light are only Bladder cancer cells have an effect and have no effect on normal cells.

The invention also provides a method for treating urinary tract epithelial cancer, wherein the method comprises the steps of: preparing a perfusate comprising an effective amount of an acridine derivative; and illuminating the perfusate with blue light. Among them, the preparation method of the perfusate and the irradiation method of the blue light have been described in the foregoing, and will not be described herein.

In summary, the present invention provides an acridine derivative for the preparation of a pharmaceutical composition for treating urinary tract epithelial cancer, when the acridine derivative is used for the detection of urinary epithelial cancer cells, if the medical staff is inspecting When the diseased cells/cancer cells are found, photodynamic therapy can be performed immediately, and the affected part is irradiated with blue light to instantly kill or reduce the number of urinary tract epithelial cancer cells during the detection of urinary tract epithelial cancer cells. In this way, the treatment can be performed at the first time when the cancer cells are detected, thereby simplifying the complicated examination and treatment process of the patient. The pharmaceutical composition can also be treated after the diagnosis of urinary tract epithelial cancer, for example, combined with bladder drug perfusion therapy, in particular, adjuvant chemotherapy or BCG treatment for urinary tract epithelial cancer to improve the treatment rate.

The above is intended to be illustrative only and not limiting. Any equivalent modifications or alterations to the spirit and scope of the invention are intended to be included in the scope of the appended claims.

B‧‧‧Bleak
C‧‧‧ catheter
K‧‧‧Kidney
O‧‧‧ Hole
Ua‧‧ urethra
Ur‧‧‧ ureter

Fig. 1 is a schematic view showing a catheter inserted into a bladder for perfusion in an experimental example of the present invention. 2A to 2D are experimental results of the effects of acridine derivatives or blue light on the activity of bladder cancer cells 5637 and T24 cells. 3A to 3C show the results of experiments on the effects of acridine derivatives and blue light on normal cells (human immortal urothelial cell line SV-Huc-1). 4A to 4D are experimental results of the effects of acridine derivatives and blue light on normal cells (human immortal urothelial cell line SV-Huc-1). 5A to 5D are experimental results of the effects of acridine derivatives and blue light on normal cells (human immortal urothelial cell line SV-Huc-1).

Claims (9)

Use of an acridine derivative for the preparation of a pharmaceutical composition for treating urinary tract epithelial cancer, wherein the pharmaceutical composition comprises an effective amount of an acridine derivative, wherein the pharmaceutical composition is irradiated with a light when treated, and the light Having an excitation peak wavelength required for the acridine derivative, wherein the acridine derivative is selected from the group consisting of acridine orange and its salts, acridine yellow and its salts, acridine red and salts thereof, 9- a group consisting of amino-6-chloro-2-methoxyacridine and salts thereof, methyl acridine and salts thereof, quinacrine and salts thereof, and any combination of the foregoing, wherein The preparation method of the pharmaceutical composition comprises: preparing a perfusate comprising an effective amount of an acridine derivative and an acidic buffer, wherein the concentration of the acridine derivative in the perfusate is greater than 1.0 μg/mL, and the perfusate The pH value is between pH 5 and pH 8. The pharmaceutical composition is perfused into at least a part of the urinary system during treatment, and the concentration of the acridine derivative in the urinary system is between 1.0 μg/mL and 5.0. Mg/mL. The use of claim 1, wherein the pharmaceutical composition is treated with blue light when treated. The use according to claim 1, wherein the acidic buffer is an acetate buffer, a monophosphate buffer, a citrate buffer, a tris buffer or a combination thereof. The use of claim 1, wherein the perfusate is perfused to at least a portion of the urinary system by perfusion. The use of claim 4, wherein the catheter is a catheter of an endoscope group, the light being emitted by the endoscope group. The use of claim 2, wherein the treatment with blue light means that at least a portion of the urinary system is illuminated by an external light source. The use of claim 2, wherein the peak wavelength of the blue light is between 400 nm and 490 nm. The use of claim 2, wherein the blue light is irradiated for more than 25 seconds. The use of claim 2, wherein the time of illuminating the blue light is between 25 seconds and 900 seconds.