A pharmaceutical composition for treating pancreatic cancer and a preparation method thereof

Abstract

This invention belongs to the field of pharmaceutical technology, and particularly relates to a pharmaceutical composition for treating pancreatic cancer and its preparation method. The pharmaceutical composition comprises, by weight, the following components: 20-25 parts gemcitabine, 0.3-0.5 parts pinocembrin, and 5-12 parts sophoridine derivative. This invention, by combining gemcitabine, sophoridine derivative, and pinocembrin, maintains synergistic antitumor activity while significantly reducing liver damage, achieving the dual goals of "enhanced efficacy and reduced toxicity."

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a pharmaceutical composition for treating pancreatic cancer and its preparation method. Background Technology

[0002] Cancer is a major global public health problem, and conquering cancer is one of the effective ways to extend human lifespan. Among the top ten malignant tumors, pancreatic cancer, due to its poor prognosis and similar mortality and morbidity rates, is the third leading cause of death for both men and women.

[0003] Clinically, based on tumor resectability, localized pancreatic cancer is classified into three main types: resectable, marginally resectable (the tumor is still localized but has surrounded or closely adhered to important blood vessels), and locally advanced unresectable. For the first two types of patients, the common practice is to perform radical surgery followed by single-agent chemotherapy such as gemcitabine to delay recurrence. For locally advanced cases where surgery is no longer an option, systemic treatment with gemcitabine combined with albumin-bound paclitaxel is commonly used. However, pancreatic cancer cells often develop rapid and refractory resistance to gemcitabine early in treatment, significantly weakening the efficacy and making the disease more difficult to control, directly increasing mortality. In addition, gemcitabine itself has clear hepatotoxicity and nephrotoxicity, limiting its long-term use and making it difficult to meet the needs of continuous treatment for pancreatic cancer patients.

[0004] Therefore, there is an urgent need for a pharmaceutical composition for the treatment of pancreatic cancer to overcome the shortcomings of existing treatments. Summary of the Invention

[0005] In order to overcome the shortcomings of the prior art, one of the objectives of the present invention is to provide a pharmaceutical composition for treating pancreatic cancer with excellent anti-tumor activity.

[0006] One of the objectives of this invention is achieved through the following technical solution:

[0007] A pharmaceutical composition for treating pancreatic cancer, comprising, by weight, the following components: 20-25 parts gemcitabine, 0.3-0.5 parts pinoside, and 9-12 parts sophoridine derivative; the structure of the sophoridine derivative is as follows:

[0008] .

[0009] Furthermore, the preparation process of the sophoridine derivative includes the following steps:

[0010] (1) 3-methoxy-4,5-ethylenedioxybenzaldehyde, sophoridine and 4-dimethylaminopyridine were added to a mixed solution of tetrahydrofuran and water to react. After the reaction was completed, the mixture was purified to obtain intermediate 1.

[0011] The structural formula of intermediate 1 is as follows:

[0012]

[0013] (2) The intermediate 1, cesium carbonate and tetrahydrofuran were mixed and stirred evenly. Then 1-methyl-3-bromo-2-piperidinone was added to react. After the reaction was completed, the product was purified to obtain the sophoridine derivative.

[0014] Furthermore, the molar ratio of 3-methoxy-4,5-ethylenedioxybenzaldehyde, sophoridine, and 4-dimethylaminopyridine in step (1) is 1:(1.2~1.5):(0.2~0.5).

[0015] Furthermore, the reaction time in step (1) is 16-18 hours.

[0016] Furthermore, the volume ratio of tetrahydrofuran to water in step (1) is 1:1.

[0017] Furthermore, in step (2), the molar ratio of intermediate 1, cesium carbonate, and 1-methyl-3-bromo-2-piperidinone is 1:(1~1.4):(1.2~1.6).

[0018] Furthermore, the reaction time in step (2) is 2 to 5 hours.

[0019] The second objective of this invention is to provide a method for preparing a pharmaceutical composition for treating pancreatic cancer, which has a simple preparation process.

[0020] The second objective of this invention is achieved by the following technical solution:

[0021] The preparation method of the above-mentioned pharmaceutical composition for treating pancreatic cancer includes the following steps:

[0022] Gemcitabine, pinocembrin, and sophoridine derivatives were weighed according to the stated weight ratio, mixed evenly, and the pharmaceutical composition for treating pancreatic cancer was obtained.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0024] This invention utilizes the Morita-Baylis-Hillman reaction to condense sophoridine with 3-methoxy-4,5-ethylenedioxybenzaldehyde, yielding a novel sophoridine derivative. This derivative can reverse the gemcitabine resistance in pancreatic cancer cells, providing a potential strategy for overcoming the clinical challenge of chemotherapy resistance in pancreatic cancer. Further research revealed that combining gemcitabine, the sophoridine derivative, and coniferin significantly reduces liver damage while maintaining synergistic antitumor activity, thus achieving the dual goals of "enhancing efficacy and reducing toxicity." Detailed Implementation

[0025] The present invention will be further described below with reference to specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments. Specific conditions not specified in the embodiments shall be performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, all reagents or instruments used are conventional products obtained through commercial channels.

[0026] (a) Preparation example

[0027] Preparation Example 1

[0028] Preparation Example 1 provides a sophoridine derivative, and the preparation process is as follows:

[0029]

[0030] (1) Add 3-methoxy-4,5-ethylenedioxybenzaldehyde (CAS: 75889-54-2, 10 mmol), sophoridine (CAS: 6483-15-4, 13 mmol), and 4-dimethylaminopyridine (DMAP, 4 mmol) to a tetrahydrofuran / water mixture (10 mL, V 四氢呋喃 V 水 The reaction mixture (r=1:1) was reacted at room temperature for 17 h. After the reaction was complete, the reaction solution was diluted with water, then extracted with dichloromethane. The combined organic layers were dried with magnesium sulfate, concentrated under reduced pressure, and subjected to silica gel chromatography (V). 正己烷 V 乙酸乙酯 Elution with a ratio of 1:1 yielded intermediate 1; the NMR and mass spectrometry results of intermediate 1 are as follows:

[0031] 1 HNMR (C 25 H 32 N₂O₅, 400MHz, DMSO-d₆)δ 6.66(s, 2H), 5.73(t, 1H), 5.59(s, 1H), 5.13(s, 1H), 4.29(s, 4H), 3.73(s, 3H), 3.48-3.46(m, 1H), 3.35-3.33(m, 1H), 3.23-3.21(m, 1H), 2.49-2.30(m, 5H), 2.06-2.04(m, 1H), 1.69(t, 1H), 1.56-1.31(m, 10H); MS m / z 441.23(M+1), found: 441.25.

[0032] (2) The above intermediate 1 (0.25 mmol), cesium carbonate (CAS: 534-17-8, 0.3 mmol), and tetrahydrofuran (2 mL) were mixed and stirred evenly at room temperature. Then, 1-methyl-3-bromo-2-piperidinone (CAS: 49785-85-5, 0.35 mmol) was added and reacted for 4 h. After the reaction was completed, the reaction solution was filtered through a diatomaceous earth layer, eluted with tetrahydrofuran, and extracted with ethyl acetate. The organic layers were combined, washed with water, dried over magnesium sulfate, concentrated under reduced pressure, and subjected to silica gel chromatography (V) 正己烷 V 乙酸乙酯 The sophoridine derivative was obtained by elution with a ratio of 3:1; the NMR and mass spectrometry results of the sophoridine derivative are as follows:

[0033] 1 HNMR (C 31 H 41 N3O6,400MHz,DMSO-d6)δ6.66(s,2H),5.73(t,1H),5.29(s,1H),4.29(s,4H),4.15(t,1H),3.73(s,3H),3.48-3.46(m,1H),3 MS m / z551.30(M+1), found: 551.31.

[0034] Preparation Example 2

[0035] Preparation Example 2 provides a sophoridine derivative, and the preparation process is as follows:

[0036] (1) Add 3-methoxy-4,5-ethylenedioxybenzaldehyde (10 mmol), sophoridine (12 mmol), and 4-dimethylaminopyridine (DMAP, 2 mmol) to a tetrahydrofuran / water mixture (10 mL, V 四氢呋喃 V 水 The reaction mixture (r=1:1) was reacted at room temperature for 16 h. After the reaction was complete, the reaction solution was diluted with water, then extracted with dichloromethane. The combined organic layers were dried with magnesium sulfate, concentrated under reduced pressure, and subjected to silica gel chromatography (V). 正己烷 V 乙酸乙酯 =1:1) Elution and separation to obtain intermediate 1; the NMR and mass spectrometry results of intermediate 1 are the same as those in preparation example 1;

[0037] (2) The above intermediate 1 (0.25 mmol), cesium carbonate (0.25 mmol), and tetrahydrofuran (2 mL) were mixed and stirred evenly at room temperature. Then, 1-methyl-3-bromo-2-piperidinone (0.3 mmol) was added and the mixture was reacted for 2 h. After the reaction was completed, the reaction solution was filtered through a diatomaceous earth layer, eluted with tetrahydrofuran, and extracted with ethyl acetate. The organic layers were combined, washed with water, dried over magnesium sulfate, concentrated under reduced pressure, and subjected to silica gel chromatography (V). 正己烷 V 乙酸乙酯 The sophoridine derivative was obtained by elution with a ratio of 3:1. The NMR and mass spectrometry results of the sophoridine derivative were the same as those in Preparation Example 1.

[0038] Preparation Example 3

[0039] Preparation Example 3 provides a sophoridine derivative, and the preparation process is as follows:

[0040] (1) Add 3-methoxy-4,5-ethylenedioxybenzaldehyde (10 mmol), sophoridine (15 mmol), and 4-dimethylaminopyridine (DMAP, 5 mmol) to a tetrahydrofuran / water mixture (10 mL, V) 四氢呋喃 V 水 The reaction mixture was prepared at a 1:1 ratio and reacted at room temperature for 18 h. After the reaction was complete, the reaction solution was diluted with water and then extracted with dichloromethane. The combined organic layers were dried over magnesium sulfate, concentrated under reduced pressure, and subjected to silica gel chromatography (V). 正己烷 V 乙酸乙酯 =1:1) Elution and separation to obtain intermediate 1; the NMR and mass spectrometry results of intermediate 1 are the same as those in preparation example 1;

[0041] (2) The above intermediate 1 (0.25 mmol), cesium carbonate (0.35 mmol), and tetrahydrofuran (2 mL) were mixed and stirred evenly at room temperature. Then, 1-methyl-3-bromo-2-piperidinone (0.4 mmol) was added and the mixture was reacted for 5 h. After the reaction was completed, the reaction solution was filtered through a diatomaceous earth layer, eluted with tetrahydrofuran, and extracted with ethyl acetate. The organic layers were combined, washed with water, dried over magnesium sulfate, concentrated under reduced pressure, and subjected to silica gel chromatography (V) 正己烷 V 乙酸乙酯 The sophoridine derivative was obtained by elution with a ratio of 3:1; the NMR and mass spectrometry results of the sophoridine derivative were the same as those in Preparation Example 1.

[0042] (II) Implementation Examples

[0043] Example 1

[0044] This embodiment provides a pharmaceutical composition for treating pancreatic cancer, comprising the following components by weight: 22 parts gemcitabine, 0.4 parts pinoside, and 10 parts of the sophoridine derivative prepared in Preparation Example 1.

[0045] This embodiment also provides a method for preparing the above-mentioned pharmaceutical composition for treating pancreatic cancer as follows:

[0046] Gemcitabine, pinocembrin, and sophoridine derivatives were weighed according to the above-mentioned weight ratio, mixed evenly, and the pharmaceutical composition for treating pancreatic cancer was obtained.

[0047] Example 2

[0048] This embodiment provides a pharmaceutical composition for treating pancreatic cancer, comprising the following components by weight: 25 parts gemcitabine, 0.5 parts pinoside, and 12 parts sophoridine derivative prepared in Preparation Example 2.

[0049] This embodiment also provides a method for preparing the above-mentioned pharmaceutical composition for treating pancreatic cancer as follows:

[0050] Gemcitabine, pinocembrin, and sophoridine derivatives were weighed according to the above-mentioned weight ratio, mixed evenly, and the pharmaceutical composition for treating pancreatic cancer was obtained.

[0051] Example 3

[0052] This embodiment provides a pharmaceutical composition for treating pancreatic cancer, comprising the following components by weight: 20 parts gemcitabine, 0.3 parts pinoside, and 9 parts of the sophoridine derivative obtained in Preparation Example 3.

[0053] This embodiment also provides a method for preparing the above-mentioned pharmaceutical composition for treating pancreatic cancer as follows:

[0054] Gemcitabine, pinocembrin, and sophoridine derivatives were weighed according to the above-mentioned weight ratio, mixed evenly, and the pharmaceutical composition for treating pancreatic cancer was obtained.

[0055] (III) Comparative Example

[0056] Comparative Example 1

[0057] This comparative example provides a pharmaceutical composition comprising, by weight, the following components: 22 parts gemcitabine and 10 parts sophoridine derivative prepared in Preparation Example 1.

[0058] Comparative Example 2

[0059] This comparative example provides a pharmaceutical composition comprising, by weight, the following components: 22 parts gemcitabine, 10 parts sophoridine, and 0.4 parts pinoside.

[0060] (iv) Test Cases

[0061] Experimental Example 1

[0062] (a) with 1×10 4At a cell density of cells / mL, drug-resistant pancreatic cancer Panc-1 cells were seeded in 35 mm culture dishes and cultured at 37°C and 5% CO2 for 24 h for later use.

[0063] (b) Gemcitabine solutions with concentrations of 1 μM, 5 μM, 10 μM, 20 μM, 40 μM, and 80 μM, and sophoridine derivative solutions with concentrations of 0 μM, 5 μM, 10 μM, and 20 μM, were prepared using α-MEM medium (containing 1% fetal bovine serum). The two solutions were then applied in combination to the drug-resistant Panc-1 cells from step (a). After 72 h of combined treatment, the effect of different drug concentrations on cell growth inhibition was determined and analyzed using the trypan blue exclusion assay. The results are shown in Table 1-2.

[0064] Table 1

[0065]

[0066] Table 2

[0067]

[0068] As shown in Table 1, compared with gemcitabine alone, the combination of gemcitabine and sophoridine derivatives reduced the resistance of resistant Panc-1 cells to gemcitabine. When gemcitabine was used in combination with a sophoridine derivative at a concentration of 20 μM, the growth inhibition effect on resistant Panc-1 cells was the best.

[0069] As shown in Table 2, compared with gemcitabine alone, the combination of gemcitabine and sophoridine derivatives significantly reversed the resistance of drug-resistant Panc-1 cells to gemcitabine and significantly increased the resistance reversal fold.

[0070] Experimental Example 2

[0071] 1. Laboratory animals

[0072] A total of 70 BALB / c mice were used, with a weight of 18-22g.

[0073] 2. Model Building

[0074] Pan02 mouse pancreatic cancer cell line in logarithmic growth phase was obtained and diluted to 1×10⁻⁶ cells using α-MEM medium (containing 10% FBS and 1% penicillin-streptomycin). 7 / mL. Under sterile conditions, 100 μL of Pan02 cell solution was injected subcutaneously into the axillary region of mice. The mice were observed daily after inoculation, and the long diameter (a) and short diameter (b) of the tumor were measured using calipers. The tumor volume (mm²) was calculated according to the formula. 3 )=(a×b 2 Calculate the tumor volume by 2, and continue until the tumor volume reaches 200 mm. 3Treatment will begin in groups at that time.

[0075] 3. Experimental Grouping

[0076] All mice were randomly divided into 7 groups: model group, gemcitabine group, and experimental group (divided into Example 1-3 groups and Comparative Example 1-2 groups), with 10 mice in each group.

[0077] 4. Administration method

[0078] Model group: Orally administered an equal volume of normal saline, 3 times a day, for 30 days;

[0079] Gemcitabine (Gem) group: Gemcitabine was administered intraperitoneally at a dose of 50 mg / kg, 3 times a day, for 30 days.

[0080] Example 1 group: The composition of Example 1 group was administered intraperitoneally at a dose of 30 mg / kg, 3 times a day, for 30 days;

[0081] Example 2 group: The composition of Example 2 group was administered intraperitoneally at a dose of 30 mg / kg, 3 times a day, for 30 days;

[0082] Example 3 group: The composition of Example 3 group was administered intraperitoneally at a dose of 30 mg / kg, 3 times a day, for 30 days;

[0083] Comparative Example 1: The drug of Comparative Example 1 was administered intraperitoneally at a dose of 30 mg / kg, once every 3 days for 30 days.

[0084] Comparative Group 2: The drug of Comparative Group 2 was administered intraperitoneally at a dose of 30 mg / kg, once every 3 days for 30 days.

[0085] Twenty-four hours after the last administration, mice in each group were sacrificed by cervical dislocation, and the tumor masses were removed and weighed. The tumor inhibition rate of each group was calculated as follows: Tumor inhibition rate = (tumor mass of model group - tumor mass of experimental group) / tumor mass of model group × 100%. The results are shown in Table 3. The liver and kidney indices, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr) levels of mice in each group were measured. The results are shown in Table 4.

[0086] Table 3

[0087]

[0088] Table 4

[0089]

[0090] As shown in Table 3, the pharmaceutical compositions of Examples 1-3 were able to inhibit the growth of Pan02 cell xenografts in nude mice. Compared with the omission of pinocembrin (Comparative Example 1) and the use of sophoridine instead of sophoridine derivatives (Comparative Example 2), the compositions of Examples 1-3 exhibited extremely high tumor inhibition rates. This indicates that the above compositions have good efficacy in treating pancreatic cancer, and that the components have a synergistic effect.

[0091] Table 4 shows that gemcitabine alone reduced the liver and kidney coefficients in mice, while significantly increasing ALT and AST levels, indicating obvious liver damage. However, Example 1, which combined gemcitabine, a sophoridine derivative, and pinocembrin, effectively reduced liver damage while maintaining synergistic antitumor activity.

[0092] In summary, this invention utilizes the Morita-Baylis-Hillman reaction to condense sophoridine with 3-methoxy-4,5-ethylenedioxybenzaldehyde to prepare a novel sophoridine derivative. This derivative can reverse the resistance of pancreatic cancer cells to gemcitabine, providing a potential strategy for overcoming the clinical challenge of chemotherapy resistance in pancreatic cancer. Further research revealed that the combined use of gemcitabine, the sophoridine derivative, and coniferin significantly reduces liver damage while maintaining synergistic antitumor activity, thus achieving the dual goals of "enhancing efficacy and reducing toxicity."

[0093] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.

Claims

1. A pharmaceutical composition for treating pancreatic cancer, characterized in that, By weight, it comprises the following components: gemcitabine 20-25 parts, pinoside 0.3-0.5 parts, and sophoridine derivative 9-12 parts; the structure of the sophoridine derivative is as follows: 。 2. A method for preparing a sophoridine derivative, characterized in that, Includes the following steps: (1) 3-methoxy-4,5-ethylenedioxybenzaldehyde, sophoridine and 4-dimethylaminopyridine were added to a mixed solution of tetrahydrofuran and water to react. After the reaction was completed, the mixture was purified to obtain intermediate 1. The structural formula of intermediate 1 is as follows: (2) The intermediate 1, cesium carbonate and tetrahydrofuran were mixed and stirred evenly, and then 1-methyl-3-bromo-2-piperidinone was added to react; after the reaction was completed, the product was purified to obtain the sophoridine derivative. The structure of the sophoridine derivative is as follows: 。 3. The method for preparing the sophoridine derivative according to claim 2, characterized in that, The molar ratio of 3-methoxy-4,5-ethylenedioxybenzaldehyde, sophoridine, and 4-dimethylaminopyridine in step (1) is 1:(1.2~1.5):(0.2~0.5).

4. The method for preparing the sophoridine derivative according to claim 2, characterized in that, The reaction time described in step (1) is 16-18 hours.

5. The method for preparing the sophoridine derivative according to claim 2, characterized in that, The volume ratio of tetrahydrofuran to water in step (1) is 1:

1.

6. The method for preparing the sophoridine derivative according to claim 2, characterized in that, The molar ratio of intermediate 1, cesium carbonate, and 1-methyl-3-bromo-2-piperidinone in step (2) is 1:(1~1.4):(1.2~1.6).

7. The method for preparing the sophoridine derivative according to claim 2, characterized in that, The reaction time in step (2) is 2 to 5 hours.

8. A method for preparing the pharmaceutical composition for treating pancreatic cancer according to claim 1, characterized in that, Includes the following steps: Gemcitabine, pinocembrin, and sophoridine derivatives were weighed according to the stated weight ratio, mixed evenly, and the pharmaceutical composition for treating pancreatic cancer was obtained.

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