NEW INDICATION OF PAROXETINE PHARMACEUTICAL COMPOSITION FOR TREATING CANCER

Info

Publication number: 20170304286
Type: Application
Filed: Oct 23, 2015
Publication Date: Oct 26, 2017
Applicant: LAUNX BIOMEDICAL CO., LTD. (Kaohsiung)
Inventors: Chiu-Hung Chen (Kaohsiung), Show-Mei Chuang (Taichung), Tzong-Der Way (Kaohsiung), Nai-Wan Hsiao (Taichung)
Application Number: 15/521,524

Abstract

A method for treating a cancer includes administering to a subject in need thereof a pharmaceutical composition containing a therapeutically effective amount of Paroxetine or a pharmaceutical acceptable salt thereof. The cancer is selected from the group consisting of pleural-related cancer, abdominal-related cancer, endocrine-related cancer, gastrointestinal tract-related cancer, osteosarcoma, skin cancer, and blood cancer. The pleural-related cancer is lung cancer. The abdominal-related cancer is selected from bladder cancer, cervical cancer, and kidney cancer. The endocrine-related cancer is selected from prostate cancer, breast cancer, and ovarian cancer. The gastrointestinal tract-related cancer is selected from gastric cancer, hepatic cancer, colorectal cancer, pancreatic cancer, and tongue cancer.

Description

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a National Phase Application filed under 35 U.S.C. 371 as a national stage of PCT/CN2015/092780 filed Oct. 23, 2015, an application claiming the benefit under 35 USC 119(e) to the following U.S. Provisional Applications No. 62/068,298 filed Oct. 24, 2014, the content of each of which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention related to a new indication of Paroxetine pharmaceutical composition, especially related to inhibition effect of Paroxetine pharmaceutical composition on a variety of cancer cells.

BACKGROUND OF THE INVENTION

Cancer is the most popular disease cause of death in the world. The cancer patients are gradually increase yearly, therefore the treatment method of the cancer has become an important issue. The medical treatments of cancer can be classified as surgical treatment, radiation therapy, chemotherapy and target therapy. Generally, the cancer drug, whether chemotherapy drug or target therapy drug, is to inhibit cancer cells duplication and split to prevent the tumor growth and metastasis.

Recently, the drug design for cancer is mainly focused on development of high-specific drug molecules or target-based antibody. Averagely, only about five of 10,000 new drugs can successfully enter the phase I of clinical trials.

Otherwise, the manufacturing of the drug is also a big problem. When the drug starting the clinical trials, there are lots of problems need to overcome, such as drug safety, patient selection, trial dose and other issues. Even the drug has approved by the FDA and sales on the market, there still possibly face the situation of the poor drug response in patients. Furthermore, if the cancer patients happen the drug resistance, that would reduce the effectiveness of the drugs and result in the medical treatment failure. Therefore, the new drug development is very difficult.

Paroxetine, also known by the trade names Paxil and Seroxat, is an antidepressant of the selective serotonin reuptake inhibitor (SSRI) class. It is used to treat major depressive disorder, obsessive-compulsive disorder, social anxiety disorder, panic disorder, posttraumatic stress disorder, generalized anxiety disorder and premenstrual dysphoric disorder. Paroxetine is approved by FDA and accumulated a huge data of drug use and drug mechanism research.

Due to the differences of the clinical use, there is no research present that the paroxetine has any potential to inhibit cancer cell.

SUMMARY OF THE INVENTION

In order to solve the above problems, the present invention provide the development of new cancer clinical indications of Paroxetine.

Accordingly, the present invention provides a new indication of Paroxetine. The experimental results showed that the Paroxetine had no toxicity or had little toxicity to normal cells in the present invention.

The present invention provides a pharmaceutical composition of Paroxetine for treating cancer. The pharmaceutical composition is composed of effective dose of Paroxetine and a pharmaceutical acceptable salt.

In one embodiment of the present invention, the cancer is selected from pleural-related cancer, abdominal-related cancer, endocrine-related cancer, gastrointestinal tract-related cancer.

In one embodiment of the present invention, the cancer is selected from osteosarcoma, skin cancer and blood cancer.

In one embodiment of the present invention, the pleural-related cancer is lung cancer.

In one embodiment of the present invention, the abdominal-related cancer is selected from bladder cancer, cervical cancer and kidney cancer.

In one embodiment of the present invention, the endocrine-related cancer is selected from prostate cancer, breast cancer, and ovarian cancer.

In one embodiment of the present invention, the gastrointestinal tract-related cancer is selected from gastric cancer, hepatic cancer, colorectal cancer, pancreatic cancer, and tongue cancer.

In one embodiment of the present invention, the effective dose of Paroxetine is from 20 mg/kg/day to 500 mg/kg/day.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the results of the inhibitory effect of the different cancer cells by Paroxetine.

FIG. 2 shows the results of the inhibitory effect of gastric tumor volume by Paroxetine.

FIG. 3 shows the inhibitory effect of gastric tumor growth via administered high-dose and low-dose of Paroxetine.

DETAILED DESCRIPTION OF THE INVENTION

Cell Culture

Subculture the different types of cancer cells. The cancer cells includes lung cancer, gastric cancer, hepatic cancer, colon cancer, skin cancer, cervical cancer, prostate cancer, bladder cancer, breast cancer, leukemia, pancreatic cancer, ovarian cancer, tongue cancer, osteosarcoma, and renal cancer. The normal cells used in the control group included kidney cells (HEK293), canine fibroblast cell line HFW, and human bronchial epithelial cell line BEAS-2B. (as shown in Table 1).

Cancer cell lines were cultured in different culture medium according to different characteristics (as shown in Table 1). The cell numbers were counted and reseed as 2×10⁶in culture plate/flask. Then, the culture medium were added to a volume of 10 ml, and the cells were cultured for 2-3 days. Then, the cells were suspended for loading into 96-well plates. The number of cells was 3000 and the volume of the culture medium was 100 μl each well.

TABLE 1 Cancer cell lines and culture medium Culture No Cancer type Cancer cell type medium 1 lung cancer H1650 (lung adenocarcinoma) RPMI-1640 A549 (lung adenocarcinoma) DMEM 2 gastric cancer AGS (Gastric Adenocarcinoma) RPMI-1640 MKN-45 (Gastric RPMI-1640 Adenocarcinoma) 3 hepatic cancer HepG2 (hepatocellular DMEM carcinoma) DMEM Hep3B (hepatocellular carcinoma) 4 colon cancer HCT116(p53+) (colorectal DMEM carcinoma) DMEM LoVo(Colorectal Adenocarcinoma) 5 skin cancer A375 (amelanotic melanoma) DMEM BCC (basal cell carcinoma) DMEM 6 cervical cancer HeLa (CervixAdenocarcinoma) DMEM C-33A (Cervical carcinoma) MEM BCRC60554 7 prostate cancer PC3 (p53−)(Prostate DMEM adenocarcinoma) RPMI-1640 LNCaP clone FGC (LNCap.FGC) 8 bladder cancer 8301 (urinary bladder RPMI-1640 carcinoma) RPMI-1640 T24 9 breast cancer MCF7 (Mammary Gland, DMEM Adenocarcinoma) DMEM MDA-MB-231 (Mammary Gland, Adenocarcinoma) 10 pancreatic cancer BxPC-3 RPMI-1640 AsPC-1 RPMI-1640 11 ovarian cancer NIH: OVCAR-3 RPMI-1640 TOV-21G RPMI-1640 12 tongue cancer SAS (Tongue squamouscell DMEM carcinoma) 13 osteosarcoma U-2OS DMEM 14 renal cancer 786-O (Renal adenocarcinoma) RPMI-1640 BCRC 60243 15 normal cell kidney HEK293 (Kidney), MDCK, DMEM pulmonary VERO RPMI-1640 epithelial cell line BEAS-2B (Lung Epithelial)

Cell Viability Analysis

Removing the original culture medium from 96-well plate. Then add 100 μl of commercially drug at a concentration of 10 μM per well. After 72 hours, add the diluted WST-1 reagent to the well with 100 μl/well, and the diluted WST-1 reagent was acquired from the dilution of 9:1 medium and WST-1 stock reagent. Finally, the total volume of each well was 200 μl/well. Culture the 96-well plate at 37° C. for 30 to 90 minutes. Detecting and calculate the survival rate of each cancer cells with an ELISA reader at OD450 nm. The lower viability of cancer cells represents better inhibition effect via the Paroxetine drug. Otherwise, the higher viability of cancer cells represents worse inhibition effect via the Paroxetine drug.

The Effect of Paroxetine on Different Cancer Cell Lines

The Inhibition Effect of Paroxetine on Pleural-Related Cancer Cells

This inhibition test of Paroxetine on pleural-related cancer cells were using two lung cancer cell lines A549 and H1650. The inhibitory tests of Paroxetine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 2.

TABLE 2 The inhibition effect of Paroxetine on pleural-related cancer cell lines 0524-10 0526-10 0529-10 0531-10 min min min min Average A549 84.19 127.41 54.01 104.12 92.43 1-10 2-20 3-20 4-20 min min min min Average H1650 66.4 31.6 58.2 71.3 56.9

The Inhibition Effect of Paroxetine on Abdominal-Related Cancer Cell Lines

This inhibition test of Paroxetine on abdominal-related cancer cells were using bladder cancer cell lines TSGH and T24 (Table 3), cervical cancer cell lines HeLa and C-33A (Table 4), renal cancer cell line 786-O (Table 5).The inhibitory tests of Paroxetine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 3, Table 4, and Table 5.

TABLE 3 The inhibition effect of Paroxetine on bladder cancer cell lines 0510-10 0512-10 0515-10 0517-10 min min min min Average TSGH 37.09 37.39 36.75 42.18 38.4 1-30 2-20 3-20 4-20 min min min min Average T24 111.7 85.2 101.5 101.5 99.9

TABLE 4 The inhibition effect of Paroxetine on cervical cancer cell lines 0524-10 min 0526-10 min 0529-10 min 0531-10 min Average HeLa 52.61 78.32 50.99 73.36 63.82 C- 33.2 32.1 31.7 35.8 33.2 33A

TABLE 5 The inhibition effect of Paroxetine on renal cancer cell line 0524-10 min 0526-10 min 0529-10 min 0531-10 min Average 786- 35.2 24.0 14.6 22.3 24.0 O

The Inhibition Effect of Paroxetine on Endocrine-Related Cancer Cell Lines

This inhibition test of Paroxetine on endocrine-related cancer cells were using prostate cancer cell lines PC-3 and LNCap (Table 6), breast cancer cell lines MCF7 and MDA-MB-231 (Table 7),and ovarian cancer cell linesNIH-OVCAR-3 and TOV-21G(Table 8). The inhibitory tests of Paroxetine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 6, Table 7, and Table 8.

TABLE 6 The inhibition effect of Paroxetine on prostate cancer cell lines PC-3-0524- PC-3-0526- PC-3-0529- PC-3-0531- Aver- 10 min 10 min 10 min 10 min age PC-3 33.52 61.61 39.26 41.84 44.06 LNCap-1- LNCap-2- LNCap-3- LNCap-4- 10 min 20 min 20 min 20 min Average LNCap 65.9 61.7 80.9 78.3 71.7

TABLE 7 The inhibition effect of Paroxetine on breast cancer cell lines 0619- 0612-10 min 0614-10 min 0616-10 min 10 min Average MCF7 47.71 49.97 55.41 51.60 51.17 MDA- 47.94 35.88 42.04 41.18 41.76 MB-231

TABLE 8 The inhibition effect of Paroxetine on ovarian cancer cell lines 7-3-30 min 7-4-30 min 7-7-30 min Average 7-4- 30 min NIH- 80.2 81.9 83.9 97.8 86.0 OVCAR-3 4-30 min TOV-21G 93.5 79.7 89.2 86.1 87.1

The Inhibition Effect of Paroxetine on Gastrointestinal Tract-Related Cancer Cell Lines

This inhibition test of Paroxetine on gastrointestinal tract-related cancer cells were using gastric cancer cell lines AGS and MKN-45 (Table 9), hepatic cancer cell lines HepG2 and Hep3B (Table 10), colorectal cancer cell lines HCT116-wt and LoVo (Table 11), pancreatic cancer cell lines AsPC-1 and BxPC-3 (Table 12), and tongue cancer cell line SAS (Table 13).The inhibitory tests of Paroxetine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 9, Table 10, Table 11, Table 12and Table 13.

TABLE 9 The inhibition effect of Paroxetine on gastric cancer cell lines 0517- 0510-10 min 0512-10 min 0515-10 min 10 min Average AGS 12.02 11.10 12.65 12.85 12.2 MKN- 36.20 52.00 37.76 36.02 40.5 45

TABLE 10 The inhibition effect of Paroxetine on hepatic cancer cell lines 0531- 0524-20 min 0526-20 min 0529-20 min 20 min Average HepG2 66.44 58.28 50.07 41.73 54.13 0619- 0612-20 min 0614-20 min 0616-20 min 20 min Average Hep3B 69.85 67.38 66.16 60.90 66.07

TABLE 11 The inhibition effect of Paroxetine on colorectal cancer cell lines 0609- 0602-30 min 0605-10 min 0607-10 min 10 min Average HCT116- 23.78 30.62 29.66 29.47 28.38 wt 0623- 0616-10 min 0619-10 min 0621-10 min 10 min Average LoVo 54.11 51.54 77.28 48.93 57.97

TABLE 12 The inhibition effect of Paroxetine on pancreatic cancer cell lines 1-4- 1-7-3-30 min 1-7-4-30 min 1-7-7-30 min 30 min Average AsPC-1 78.8 87.3 69.3 72.5 77.0 3-4- 3-7-3-30 min 3-7-4-30 min 3-7-7-30 min 30 min Average BxPC-3 59.7 93.7 70.6 87.0 77.7

TABLE 13 The inhibition effect of Paroxetine on tongue cancer cell line 6-26-10 min 6-28-10 min 6-30-10 min 7-3-10 min Average SAS 48.20 26.74 57.97 73.09 51.50

The Inhibition Effect of Paroxetine on Other Cancer Cell Lines

This inhibition test of Paroxetine on other cancer cells were using osteosarcoma cell line U2OS (Table 14), skin cancer cell lines A375 and BCC (Table 15). The inhibitory tests of Paroxetine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 14 and Table 15.

TABLE 14 The inhibition effect of Paroxetine on osteosarcoma cancer cell line 7-3- 6-26-10 min 6-28-10 min 6-30-10 min 10 min Average U2OS 40.61 30.75 30.29 31.10 33.19

TABLE 15 The inhibition effect of Paroxetine on skin cancer cell lines 0609- 0602-30 min 0605-10 min 0607-10 min 10 min Average A375 46.75 44.40 35.38 48.77 43.83 BCC 42.13 52.62 41.11 97.05 58.23

The Experiment Design on Control Group

The Inhibition Effect of Paroxetine on Normal Cells

This inhibition test of Paroxetine on normal cells were using normal kidney cell line HEK293 (Table 16) and normal pulmonary epithelial cell lines BEAS-2B (Table 17). The inhibitory tests of Paroxetine were performed 4 times for each cell lines and then the average value of the inhibitory tests was calculated. The results were shown in Table 16 and Table 17.

TABLE 16 The inhibition effect of Paroxetine on normal kidney cell line 0609- 0602-30 min 0605-30 min 0607-30 min 30 min Average HEK293 64.91 76.80 76.05 67.85 71.40

TABLE 17 The inhibition effect of Paroxetine on normal pulmonary epithelial cell line 0517- 0510-10 min 0512-10 min 0515-10 min 10 min Average BEAS-2B 100.97 102.64 96.40 94.56 98.64

This inhibition test results of Paroxetine on all kinds of cells were shown in Table 18. It is clear that Paroxetine has a significant inhibitory effect on several cancer cell lines. As a result in the experiments of the present invention, Paroxetine has a significant inhibitory effect on various cancer cells. (FIG. 1)

TABLE 18 Summary of the Effect on different cancer cell lines by Paroxetine cancer cells Inhibitory effect lung cancer 74.64462204 bladder cancer 69.1 cervical cancer 63.82 prostate cancer 44.06 breast cancer 46.47 ovarian cancer 86.5 gastriccancer 26.3 hepatic cancer 60.10 colorectal cancer 43.17 pancreatic cancer 77.4 tongue cancer 51.50 osteosarcoma 33.19 skin cancer 51.03 renal cancer 72.5 kidney cell 71.40 pulmonary epithelial cell line 98.64

Animal Model Test of Gastric Cancer with Dose 100 mg/kg/day and 200 mg/kg/day

In this invention, the female mice were (BALB/cAnN.Cg-Foxn1^nu/CrlNarl) purchased from National Laboratory Animal Center (Taiwan). The weight of the mice were 21±1 g. These mice were subcutaneously injected with gastric cancer cells (AGS) and then put these mice into different cage sat random. The drug test experiment was divided into three groups, include “control group”, “low dose group (100 mg/kg/day)”, and “high dose group (200 mg/kg/day)”. These mice were then injected test drug intraperitoneally once daily until the tumor size reached 100 mm³. The tumor sizes and body weight were measured twice a week. The tumor sizes were measured and calculated by formula: (L×W²)/2. L represents the tumor longest length. W represents the tumor shortest diameter. The experiment results were shown in Table 19.

TABLE 19 The inhibitory effect of tumor volume via administered Paroxetine control group Tumor low concentration (100 mg/kg/day) longest volume longest weight length width volume growth weight length width volume (g) mm mm mm3 mm3 (g) mm mm mm3 First measurement A 18.5 7 7 171.5 171.5 19 7 5 87.5 B 22 8 6 144 144 19 8 6 144 C 20.5 9 8 288 288 18 6 6 108 average 201.1667 201.1667 113.1667 Second measurement A 22 7 6 126 −45.5 19 6 5 75 B 20 8 7 196 52 21 7 6 126 C 20 9 7 220.5 −67.5 20 7 6 126 average 180.8333 −20.3333 109 Third measurement A 23 9 6 162 36 20.5 7 6 126 B 20 10 8 320 124 23.5 7 4 56 C 21 11 7 269.5 49 19 7 5 87.5 average 250.5 69.667 89.833 Fourth measurement A 23 11 7 269.5 107.5 22 6 6 108 B 22 10 6 180 −140 22 4 4 32 C 23 11 8 352 82.5 20 6 5 75 average 267.1667 16.667 71.667 Fifth measurement A 22 12 8 384 114.5 22 6 6 108 B 22 11 8 352 172 20 4 5 50 C 23 12 9 486 134 20 4 4 32 average 407.33 140.167 63.333 high concentration (200 mg/kg/day) Tumor longest volume low concentration (100 mg/kg/day) weight length width volume growth Tumor volume growth mm3 (g) mm mm mm3 mm3 First measurement A 87.5 19 6 4 48 48 B 144 18.5 8 5 100 100 C 108 20 7 5 87.5 87.5 average 113.1667 78.5 78.5 Second measurement A −12.5 20 6 4 48 0 B −18 23 5 3 22.5 −77.5 C 18 20 7 5 87.5 0 average −4.16667 52.66667 −25.8333 Third measurement A 51 19 5 4 40 −8 B −70 18.5 0 0 0 −22.5 C −38.5 18.5 0 0 0 −87.5 average −19.167 13.333 −39.33 Fourth measurement A −18 20 0 0 0 −40 B −24 21 0 0 0 0 C −12.5 20 0 0 0 0 average −18.167 0 −13.33 Fifth measurement A 0 21 0 0 0 0 B 18 20 0 0 0 0 C −43 20 0 0 0 0 average −8.333 0 0

According to the results in FIG. 2, both low dose and high dose of Paroxetine had significant inhibition effect on tumor cells, and the weight of mice did not show a significant decrease during the experiment. These results indicate that both high and low doses of Paroxetine could keep the tested mice in healthy status during the treatment without death.

According to the results in FIG. 3, both low dose and high dose of Paroxetine had effectively slow down the tumor volume growth, and can also reduce the tumor volume. Especially, high doses of Paroxetine had better effect to inhibit tumor growth.

Although the present invention has been described in terms of specific exemplary embodiments and examples, it will be appreciated that the embodiments disclosed herein are for illustrative purposes only and various modifications and alterations might be made by those skilled in the art without departing from the spirit and scope of the invention as set forth in the following claims.

Claims

1. A method for treating a cancer comprising: administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of Paroxetine or a pharmaceutical acceptable salt thereof.

2. The method of claim 1, wherein the cancer is selected from the group consisting of pleural-related cancer, abdominal-related cancer, endocrine-related cancer, and gastrointestinal tract-related cancer.

3. The method of claim 1, wherein the cancer is selected from the group consisting of osteosarcoma, skin cancer, and blood cancer.

4. The method of claim 2, wherein the pleural-related cancer is lung cancer.

5. The method of claim 2, wherein the abdominal-related cancer is selected from the group consisting of bladder cancer, cervical cancer, and kidney cancer.

6. The method of claim 2, wherein the endocrine-related cancer is selected from the group consisting of prostate cancer, breast cancer, and ovarian cancer.

7. The method of claim 2, wherein the gastrointestinal tract-related cancer is selected from the group consisting of gastric cancer, hepatic cancer, colorectal cancer, pancreatic cancer, and tongue cancer.

8. The method of claim 1, wherein the effective amount of Paroxetine is from 20 mg/kg/day to 500 mg/kg/day.