KR102385617B1 - A composition for inhibiting tumor cell death and metastasis containing novel compounds Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) - Google Patents

Abstract

The present invention relates to a composition for inhibiting tumor cell death and metastasis containing novel compounds potassium all-trans retinoate (PARA) and potassium 9-cis retinoate (P9-cis RA). As a result of conducting several experiments for the purpose of application of PARA and P9-cis RA to the suppression of tumor cell death and cancer cell metastasis inside and outside the human body, it has been confirmed that potassium all-trans retinoate (Tretinoin Potassium Slat) has an effect of inhibiting apoptosis and metastasis in neuroblastoma, breast cancer, and lung tumors. In addition, potassium 9-cis retinoate (Alitretinoin Potassium salt) has been confirmed to inhibit metastasis of lung tumors and breast cancer, and thus can be used as an intravenous solvent for treatment, etc.

Description

A composition for inhibiting tumor cell death and metastasis containing novel compounds Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) {A composition for inhibiting tumor cell death and metastasis containing novel compounds Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA)}

The present invention relates to a composition for inhibiting tumor cell death and metastasis comprising the novel compounds Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA). In detail, it relates to a composition for inhibiting apoptosis and metastasis of tumor cells inside and outside the human body according to blood and direct application containing Potassium all-trans retinoate (Tretinoin Potassium Slat) and Potassium 9-cis retinoate (Alitretinoin Potassium salt) .

Potassium all-trans and 9-cis retinoate is a form in which the OH group of all-trans and 9-cis retinoic acid is removed and a K ⁺ ion is attached. All-trans retinoic acid, under the name of Tretinoin, is used as a raw material for the treatment of acute promyelocytic leukemia and acne. As a very lipophilic compound, it has never been applied directly to cancer cells or tumor cell products or applied through blood, and is generally applied in the form of oral administration.

In addition, 9-cis retinoic acid is used as a raw material for the treatment of severe hand eczema under the name of alitretinoin, and it is also a lipophilic compound and it is difficult to apply directly to cancer cells or tumor cell preparations or through blood vessels.

Patent No. 10-2271364, registered as an earlier application by the present applicant, discloses "a method for synthesizing novel compounds Potassium all-trans retinoate and Potassium 9-cis retinoate", and Registered Patent No. 10-2311539 discloses a novel compound Potassium Disclosed is a pharmaceutical composition for inhibiting blood coagulation comprising all-trans retinoate and Potassium 9-cis retinoate.

In the present invention, various experiments were conducted for the purpose of applying the Potassium all-trans retinoate and Potassium 9-cis retinoate developed by the present application to the inhibition of tumor cell death and cancer cell metastasis inside and outside the human body.

In other words, as a result of conducting a related experiment to trace the use of Potassium all-trans retinoate (Tretinoin Potassium Slat) and Potassium 9-cis retinoate (Alitretinoin Potassium salt) in the blood and direct application to the death and metastasis prevention of tumor cells, Potassium all- Trans retinoate (Tretinoin Potassium Slat) has been shown to inhibit apoptosis and metastasis in neuroblastoma, breast cancer, and lung tumors. In addition, Potassium 9-cis retinoate (Alitretinoin Potassium salt) has been confirmed to have an inhibitory effect on metastasis of lung tumor and breast cancer, and was developed as a composition for treatment, such as an intravenous solution.

Korean Registered Patent No. 10-2289190 (2021.08.06.) Korean Registered Patent No. 10-2311539 (2021.10.05.) Korean Patent No. 10-2271364 (June 24, 2021) Korea Registered Patent No. 10-1872950 (2018.06.25)

The present invention utilizes the hydrophilic part of novel compounds Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) for tumor cell death and metastasis inhibition by direct injection into blood or tumors. To provide a composition for preventing tumor cell death and metastasis by conducting related experiments for the purpose of application.

The present invention provides a pharmaceutical composition for inhibiting tumor cell death and cell metastasis, comprising as an active ingredient Potassium all-trans retinoate represented by Formula 1 or Potassium 9-cis retinoate represented by Formula 2 below.

[Formula 1]

[Formula 2]

Potassium all-trans retinoate (Tretinoin Potassium Slat) in the pharmaceutical composition for inhibiting tumor cell death and cell metastasis of the present invention was confirmed to have an apoptosis and metastasis inhibitory effect on neuroblastoma, breast cancer, and lung tumors. In addition, Potassium 9-cis retinoate (Alitretinoin Potassium salt) has been confirmed to have an inhibitory effect on metastasis of lung and breast cancer, and thus can be used as an intravenous solvent for treatment.

Potassium all-trans retinoate (Tretinoin Potassium Slat) of the present invention was confirmed to have apoptosis and metastasis inhibitory effect on neuroblastoma, breast cancer, and lung tumor, and Potassium 9-cis retinoate (Alitretinoin Potassium salt) inhibits metastasis of lung tumor and breast cancer The effect has been confirmed, and there is an effect that can be used as a composition for inhibiting tumor cell death and metastasis, and as an intravenous solvent for treatment.

1 shows the results of the MTS apoptosis test for renal cancer tumor cells Cak-1
Figure 2 shows the results of the MTS apoptosis test for liver tumor HEP 38.
3 shows the results of the MTS apoptosis test for lung tumor cells A549.
4 shows the apoptosis effect using PCR test on lung tumor cells.
5 shows a change in the expression level of genes related to apoptosis.
Figure 6 shows the test results of A549 cell wound healing assay (migration).
7 shows the cell status at intervals of 24 hours and 48 hours before, after, and after application of the drug used in the experiment.
8 shows the results of the A549 invasion assay.
9 shows the results of the MTS apoptosis test for breast cancer cells MDA-MB-231.
10 shows the results of the MDA-MB-231 apoptosis PCR test.
11 shows changes in the expression level of genes related to apoptosis.
12 shows the results of the MDA-MB-231 wound healing assay (migration).
Figure 13 shows the cell status before, after, 24 hours and 48 hours after application of the drug used in the experiment. ( ***: p value <0.0001, *: p value <0.05 = compared to control)
14 shows the results of the MTS apoptosis test for SH-SY5Y neuroblastoma cells. (*** -> P-value < 0.05 vs. Control, ** -> P-value < 0.01 vs. Control, * -> P-value < 0.001 vs. Control)
15 shows the SH-SY5Y apoptosis PCR test results.
16 shows changes in the expression level of genes related to apoptosis.
17 shows the results of the SH-SY5Y wound healing assay (migration).
18 shows the cell status at intervals of 24 hours and 48 hours before, after, and after application of the drug used in the experiment.
19 shows the results of the SH-SY5Y Invasion assay.
20 shows the results of the invasion inhibitory effect in RA (P9-cis RA, PARA) ionized with potassium and Cyclophosphamide.
21 shows a comparison result of inhibition of neuroblastoma cancer cell infiltration after 24 hours of treatment with each drug of 9-cis RA, All-trans RA, P9-cis RA, PARA and Cyclophosphamide at p value <0.05.

The present invention utilizes the hydrophilic portion of Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) developed and registered by the present applicant, and tumor cells for each part of the body by direct injection into blood or tumor Apoptosis and metastasis inhibitory effects were tested.

The types of tumors for the test of tumor cell death and metastasis inhibition effects were kidney cancer, liver cancer, breast cancer, and neuroblastoma, and the experimental method was MTS [3-(4, 5-dimethylthiazol-2-yl)-5-( 3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] was used to find the cytotoxic concentration of the drug, and it was confirmed by apoptosis PCR test, migration and invasion assay.

In order to understand the cancer cell metastasis inhibitory effect of each drug, wound healing assay (cancer cell migration degree) and invasion assay (cancer cell penetration degree), that is, the migration effect, which is the amount of inhibition of cancer cell migration, were analyzed, and the penetration power of cancer cells by the drug (chemotaxis) was analyzed. ) was measured and analyzed to measure the effect of invasion.

MTS apoptosis test for renal cancer tumor cells (Cak-1 cells)

1 shows the results of the MTS apoptosis test for renal cancer tumor cells Cak-1. Generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and a novel compound Potassium all-trans retinoate (PARA) for MTS apoptosis test on renal cancer tumor cell Cak-1 ) and Potassium 9-cis retinoate (P9-cis RA) were treated at concentrations of 100uM, 50uM, 25uM, and 12.5uM, respectively, and ONE-WAY ANOVA statistical analysis was performed 24 hours later.

Experimental results As shown in the results of FIG. 1 , all four drugs were confirmed and analyzed to have no effect on cell viability. From the experimental results, it was determined that all of 9-cis RA, P9-cis RA, All-trans RA, and PARA had no apoptotic effect on renal cancer cells. Therefore, drug migration and invasion assays were not performed.

MTS apoptosis test for liver cancer tumor cells (HEP 3B)

2 shows the results of the MTS apoptosis test for HEP 38 liver cancer tumor cells. Generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and novel compounds Potassium all-trans retinoate (PARA), Potassium for MTS apoptosis test for liver tumor HEP 38 9-cis retinoate (P9-cis RA) was treated at concentrations of 100uM, 50uM, 25uM, and 12.5uM, respectively, and ONE-WAY ANOVA statistical analysis was performed 24 hours later.

As a result of the experiment, it was confirmed that all four drugs had no effect on cell viability as shown in FIG. 2 . According to the experimental results, 9-cis RA, P9-cis RA, all-trans RA, and PARA were all judged to have no apoptotic effect on hepatocellular carcinoma cells.

MTS apoptosis test for lung tumor cells (A549)

3 shows the results of the MTS apoptosis test for lung tumor cells A549 (* -> P-value <0.05, ** -> P-value <0.01, *** -> P-value <0.001). Generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and novel compounds Potassium all-trans retinoate (PARA), Potassium for MTS apoptosis test on lung tumor cell A549 9-cis retinoate (P9-cis RA) was applied at concentrations of 100 uM, 50 uM, 25 uM, and 12.5 uM, respectively, and TWO-WAY ANOVA statistical analysis was performed 24 hours later.

As a result of the experiment, it was confirmed that cell viability was significantly decreased in all three drugs except for 9-cis RA as shown in FIG. 3, and it was confirmed that the effect of PARA 100uM was the best among them. From the experimental results, it is judged that PARA (Potassium all-trans retinoate) has an apoptosis effect on lung tumor cells.

Lung tumor cell A549 Apoptosis PCR test

4 shows the apoptosis effect using PCR test on lung tumor cells. As the PARA (Potassium all-trans) was confirmed through the MTS result, the apoptosis effect was confirmed through a PCR test on apoptosis.

The cells are the same as the A549 lung cancer cell line as the MTS test cell line, and the applied drugs are also the generic drugs 9-cis retinoic acid (9-cis RA), all-trans retinoic acid (All-trans RA) and the new compound Potassium all-trans 100uM of retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) was applied for 24 hours.

PCR primers Bax, Bcl-2, Caspase-3, Beta-actin disclosed in SEQ ID NOs: 1 to 8 were used, and Primer sequence information is shown in Table 1.

Sequence information of PCR Primer Bax, Bcl-2, Caspase-3, Beta-actin BCL-2 F: 5'- CCT GTG GAT GAC TGA GTA CC -3' SEQ ID NO: 1 R: 5'-ACT TGT GGC TCA GAT AGG C -3' SEQ ID NO: 2 BAX F: 5'-CTG GAC ATT GGA CTT CCT C -3' SEQ ID NO: 3 R: 5'- CTT CTT CCA GAT GGT GAG TG-3' SEQ ID NO: 4 Caspase-3 F: 5'-GAG GGG ATC GTT GTA GAA GT-3' SEQ ID NO: 5 R: 5'-ACC AGG TGC TGT GGA GTA T-3' SEQ ID NO: 6 beta-actin F: 5'-GGA CCT GAC TGA CTA CCT CA-3' SEQ ID NO: 7 R: 5'- AGC TCG TAG CTC TTC TCC A-3' SEQ ID NO: 8

Genes related to apoptosis are typically known as Bax, Bcl-2, and Caspase-3 genes. When apoptosis occurs, bax and caspase-3 genes are increased, whereas bcl-2 genes are decreased. As a result of the experiment, the expression level of bcl-2, a gene that suppresses apoptosis, was decreased in the PARA drug, but the expression level of caspase-3, which plays the most decisive role in apoptosis, did not change.

5 shows changes in the expression level of genes related to lung tumor cell death. According to the experimental results, the apoptosis test result of A549 through drug treatment of the lipophilic form of Retinoic acid and the hydrophilic form of Potassium Retinoate is not related to apoptosis, but it is judged to be inhibition of cancer or tumor cell growth.

As a result of the above experiment, based on the determination of inhibition of cancer or tumor cell growth, the center of the cell plate was scraped with a tip in the wound healing assay method, and then after the drug, an experiment to check the movement of cells through the area filled with cells over time The degree of phosphorus migration was measured.

Lung tumor cell A549 cell wound healing assay (migration) experiment

Figure 6 shows the test results of A549 cell wound healing assay (migration). Experimental results: Generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and new compounds Potassium all-trans retinoate (PARA), Potassium 9-cis retinoate (P9-cis RA) ) Before and after drug application, the cell status was checked at intervals of 24 hours and 48 hours, and it was confirmed that the speed of filling the area was slower in the cells treated with the drug compared to the filling speed of the control.

7 shows the cell status at intervals of 24 hours and 48 hours before, after, and after application of the drug used in the experiment. As a result of confirming that the cells were filled at intervals of 24 hours and 48 hours after drug treatment, it was confirmed that the cell migration rate was significantly lowered after 48 hours of drug treatment. Among lipophilic drugs, All-trans RA and hydrophilic drugs It is judged that PARA and P9C-cis RA have relatively high efficacy in inhibiting cell migration of lung tumor cells (A549). In addition, it is judged that P9-cis RA has a relatively higher inhibitory effect than PARA among hydrophilic preparations as intravenous drugs.

Since lipophilic drugs are very difficult to deliver through venous blood, there is no other application method other than direct injection of the drug into lung tumor cells. However, only hydrophilic drugs, that is, Potassium all-trans retinoate and Potassium 9-cis retinoate, are judged to be drugs that can be applied through blood.

According to the above experiment, it was confirmed that PARA and P9-cis RA as drugs can be applied as an intravenous solution for the treatment of lung tumor cells.

In addition, in addition to the wound healing assay described above, in order to check the degree of inhibition of movement for cancer cell metastasis, the cells were mixed with serum-free media and then put into a trans-well with an 8um size pour, and serum was placed under the trans-well. Invasion assay was performed as an experiment to check the extent to which cells move from the serum-free side to the active side by chemotaxis (a phenomenon in which organisms move by a chemical reaction) when the media containing this is added.

Lung tumor cell A549 invasion assay experiment

8 shows the results of the A549 invasion assay. As a result of the experiment, it was confirmed that the drugs PARA and P9-cis RA were effective in the Invasion assay as above. From the above experiment, it is judged that the efficacy of P9-cis is better among the two drugs shown in the Invasion assay.

Summarizing the above, it was confirmed that the drug P9C-cis RA had the effect of inhibiting lung tumor cell growth and metastasis.

MTS apoptosis test for breast cancer cells (MDA-MB-231)

9 shows the results of the MTS apoptosis test for breast cancer cells MDA-MB-231 (* -> P-value <0.05, ** -> P-value <0.01, *** -> P-value <0.001) Generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and the novel compound Potassium all-trans retinoate (PARA) for MTS apoptosis test on breast cancer cells MDA-MB-231 ) and Potassium 9-cis retinoate (P9-cis RA) were treated at concentrations of 100uM, 50uM, 25uM, and 12.5uM, respectively, and 24 hours later, TWO-WAY ANOVA statistical analysis was performed.

As a result of the experiment, it was confirmed that cell viability appeared at 100uM, 25uM, and 12.5uM concentrations in the Potassium all-trans RA drug as shown in FIG. 9, and there was no significant difference in cell viability and cell viability in the remaining three drugs. Therefore, according to the results of MTS, an apoptosis PCR test was performed by applying 100 uM.

Breast cancer cell MDA-MB-231 Apoptosis PCR test

10 shows the results of apoptosis PCR test of breast cancer cells MDA-MB-231. The cells are the MTS test cell line, which is the same as the MDA-MB-231 breast cancer cell line, and the applied drugs are also generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and new compounds. Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) were applied in an amount of 100 uM for 24 hours, and MDA-MB-231 (breast cancer cell) cells were treated with each drug, and then the apoptosis was reduced. Expression patterns of related genes were confirmed.

PCR Primer, Bcl-2, Bax, Caspase-3, Beta-actin identical to SEQ ID NOs: 1 to 8 of Table 1 were used. Genes related to apoptosis are typically known as Bax, Bcl-2, and Caspase-3 genes, and when apoptosis occurs, bax and caspase-3 genes are increased and, conversely, bcl-2 genes are decreased.

As a result of the experiment, it was confirmed that the expression of the gene bax causing apoptosis was increased in all four drugs, and all-trans RA and PARA were higher. The expression level of caspase-3, a gene that causes apoptosis, was not significantly increased in both drugs, 9-cis RA and P9-cis RA. It was confirmed that the expression level of the all-trans RA and PARA drugs was highly increased. In the case of bcl-2, a gene that inhibits apoptosis, it was confirmed that the expression levels of all-trans RA and PARA drugs were reduced compared to the control group and 9-cis RA and P9-cis RA.

11 shows changes in the expression level of genes related to apoptosis. As a result of the experiment, it can be judged that all-trans RA and PARA are effective in apoptosis of breast cancer, and among them, PARA is judged to have a higher apoptosis effect. In addition, when the drug is delivered through a blood vessel such as a vein, it can be delivered as the compound PARA.

Through the above experiments, it was confirmed that PARA had an apoptotic effect on breast cancer. In addition, as a hydrophilic drug, the drug can be delivered to the breast cancer site through a vein, and it can be used as an injection that can be directly injected into the breast cancer site.

Breast cancer cell MDA-MB-231 wound healing assay (migration) experiment

12 shows the results of breast cancer cell MDA-MB-231 wound healing assay (migration) (***: p value <0.0001, *: p value <0.05 = compared to control). After scraping the center of the cell plate with a tip, the wound healing assay, which is an experiment to check the movement of cells through the area filled with cells over time after the drug, was measured, that is, the degree of migration.

As a result of the experiment, the significance of 9-cis RA, p9-cis RA, and PARA compared to controls was confirmed 24 hours after drug application. In addition, the significance of 9-cis RA, p9-cis RA, and All-trans RA was confirmed compared to controls 48 hours after application of the drug.

Figure 13 shows the cell status before, after, 24 hours and 48 hours after the application of the drug used in the experiment. ( ***: p value <0.0001, *: p value <0.05 = compared to control) Experimental result p value After 48 hours at <0.05, it was confirmed that the efficacy of P9-cis RA was higher when the significance of 9-cis and P9-cis RA was confirmed. In addition, when comparing all-trans RA and PARA at p value <0.05, the significance of All-trans RA and PARA was confirmed after 24 hours (it was confirmed that the efficacy of PARA was higher after 24 hours. That is, all-trans and It was confirmed that Potassium All-trans and 9-cis Retinoate were more effective in inhibiting cancer cell migration than 9-cis Retinoic acid.

Through the above experiment, it was confirmed that P9-cis RA had an inhibitory effect on the metastasis of breast cancer cells.

MTS apoptosis test for neuroblastoma cells (SH-SY5Y)

14 shows the results of the MTS apoptosis test for neuroblastoma cells SH-SY5Y (*** -> P-value < 0.05 vs. Control, ** -> P-value < 0.01 vs. Control, * -> P -value < 0.001 vs. Control). Generic drugs 9-cis retinoic acid (9-cis RA), All-trans retinoic acid (All-trans RA) and positive control Cyclophosphamide for MTS apoptosis test for neuroblastoma cell SH-SY5Y are currently used as generic drugs Neuroblastoma treatment and novel compounds Potassium all-trans retinoate (PARA) and Potassium 9-cis retinoate (P9-cis RA) were applied at concentrations of 100uM, 90uM, 80uM, 70uM, 60uM, and 50uM, respectively, and after 24 hours, ONE-WAY ANOVA statistical analysis was performed.

As a result of the experiment, it was confirmed that both Potassium 9-cis and All-trans RA were significantly different up to the low concentration, and in the case of cyclophosphamide as a positive control, the significance was confirmed at 100uM and 90uM, both concentrations. However, in the case of P9-cis RA and PARA, there was a difference in significance at 70uM and 60uM, respectively, and SH-SY5Y had a higher cytotoxic effect than cyclophosphamide.

Neuroblastoma cell SH-SY5Y Apoptosis PCR test

15 shows the SH-SY5Y apoptosis PCR test results. Genes related to apoptosis are typically known as Bax, Bcl-2, and Caspase-3 genes. When apoptosis occurs, bax and caspase-3 genes are increased, whereas bcl-2 genes are decreased.

PCR Primer, Bcl-2, Bax, Caspase-3, Beta-actin identical to SEQ ID NOs: 1 to 8 of Table 1 were used.

16 shows changes in the expression level of genes related to apoptosis. As a result of the experiment, the expression levels of bcl-2 and Caspase-3 mRNA, which are genes that inhibit apoptosis, were increased in PARA and Cyclophosphamide drugs, and PARA decreased more in BCL-2 than Cyclophosphamide.

In addition, PARA and Cyclophosphamide had similar levels of expression of BAX and Caspase-3 mRNA, and BCL-2 mRNA decreased more with PARA than Cyclophosphamide. Therefore, PARA had a relatively higher apoptosis effect than Cyclophosphamide, which is currently used as a treatment for neuroblastoma.

Through the above experiment, it was confirmed that PARA is effective in apoptosis of neuroblastoma.

Neuroblastoma cells SH-SY5Y wound healing assay (migration)

17 shows the results of the SH-SY5Y wound healing assay (migration). As a result of the experiment, all drugs except Na (Sodium All-trans retinoic acid) were effective, and the drugs with potassium (All-trans & 9-cis retinoic acid) had higher anti-movement efficacy, and Cyclo (Cyclophosphamide) and In comparison, the migration effect was higher.

18 shows the cell status at intervals of 24 hours and 48 hours before, after, and after application of the drug used in the experiment. As a result of the experiment, when PARA (Potassium All-trans retinoate) and Cyclo (Cyclophosphamide) were compared at p value <0.05 after 24 hours, it was confirmed that Potassium All-trans Retinoate had the highest effect of inhibiting cancer cell migration. That is, it was confirmed that the cancer metastasis inhibitory effect of SH-SY5Y neuroblastoma cells was the highest.

Neuroblastoma cell SH-SY5Y Invasion assay

It was confirmed through the migration assay that PARA inhibited the metastasis of SH-SY5Y cancer cells, and the Invasion assay was performed to confirm the degree of penetration of the drug to inhibit the metastasis of cancer cells. For the experimental method, after mixing cancer cells with serum-free media, put the cells into a trans-well with an 8um size pour, and put media containing serum under the trans-well, chemotaxis (a phenomenon in which organisms move by chemical reaction) This is an experiment using the property of cells to move from the serum-free side to the active side. Drug treatment time and invasion time: 24 hours were set, and 100uM of Control, DMSO, 9-cis RA, P9-cis RA, All-trans RA, PARA, Na, Cyclophosphamide was applied as confirmed by MTS.

19 shows the results of the SH-SY5Y Invasion assay. As a result of the experiment, potassium-ionized RA (P9-cis RA, PARA) and Cyclophosphamide had an invasion inhibitory effect, and among them, PARA showed better inhibitory effect. When cyclophosphamide and PARA were compared, PARA showed higher penetration inhibition efficacy.

Figure 20 shows the results of the invasion inhibition effect in RA (P9-cis RA, PARA) and cyclophosphamide ionized with potassium. 21 shows a comparison result of inhibition of neuroblastoma cancer cell infiltration after 24 hours of treatment with each drug of 9-cis RA, All-trans RA, P9-cis RA, PARA and Cyclophosphamide at p value <0.05.

As a result of the experiment, when the inhibition of neuroblastoma cancer cell infiltration was compared after 24 hours of treatment with each of 9-cis RA, All-trans RA, P9-cis RA, PARA and Cyclophosphamide at p value <0.05, PARA in the ionized RA type It was confirmed that the inhibitory power was the highest, and it was confirmed that PARA had a higher penetration inhibitory effect when compared to PARA and Cyclophosphamide, a generic drug for neuroblastoma.

As a result of the above experiment, it was confirmed that PARA had an inhibitory effect on cancer metastasis by inhibiting the migration and invasion of neuroblastoma cancer cells through the migration and invasion assay.

Potassium all-trans retinoate (Tretinoin Potassium Slat) of the present invention was confirmed to have an apoptosis and metastasis inhibitory effect on neuroblastoma, breast cancer, and lung tumors, and Potassium 9-cis retinoate (Alitretinoin Potassium salt) was used to metastasize lung and breast cancer. Since the inhibitory effect has been confirmed, it can be used as a therapeutic intravenous solution, etc., and thus has potential industrial application.

Claims (3)

As a composition comprising Potassium all-trans retinoate represented by the following Chemical Formula 1 as an active ingredient,
The composition is a pharmaceutical composition, characterized in that it has an effect on any one tumor cell death selected from neuroblastoma, breast cancer, and lung tumor, and an inhibitory effect on cell metastasis of any one selected from neuroblastoma, breast cancer, and lung tumor.
[Formula 1]

As a composition comprising Potassium 9-cis retinoate represented by the following Chemical Formula 2 as an active ingredient,
The composition is a pharmaceutical composition, characterized in that it has an effect on killing lung tumor or neuroblastoma and inhibiting cell metastasis of breast cancer or lung tumor
[Formula 2]

The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is prepared in any one form selected from tablets, capsules, soft capsules, granules, liquids, and injections.

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