Use of pharmaceutical composition comprising retinoic acid in preparing drug for treating hepatocellular carcinoma accompanied by abdominal cavity metastasis

Abstract

Use of a pharmaceutical composition comprising retinoic acid in preparing a drug for treating hepatocellular carcinoma accompanied by abdominal cavity metastasis. The pharmaceutical composition comprises an FOLFOX4 chemotherapy regimen and retinoic acid. The FOLFOX4 chemotherapy regimen is a primary active ingredient for treating hepatocellular carcinoma accompanied by abdominal cavity metastasis. The retinoic acid is an auxiliary ingredient used for enhancing the FOLFOX4 chemotherapy regimen in treating hepatocellular carcinoma accompanied by abdominal cavity metastasis. The FOLFOX4 chemotherapy regimen consists of oxaliplatin, 5-fluorouracil, and calcium folinate. In the aspect of treating hepatocellular carcinoma accompanied by abdominal cavity metastasis, the pharmaceutical composition of the present invention exhibits significantly superior efficacy compared to equivalent doses of the FOLFOX4 chemotherapy regimen alone and the retinoic acid alone, indicating that the combined use of the two has a significant synergistic effect.

Description

Use of a pharmaceutical composition containing retinoic acid in preparing a medicament for treating hepatocellular carcinoma with abdominal metastasis

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of Chinese Application No. 2023115917737, filed on November 27, 2023. Said application No. 2023115917737 is hereby incorporated by reference in its entirety. Technical Field

[0003] The present invention belongs to the field of medicine, and particularly relates to use of a pharmaceutical composition containing retinoic acid in preparing a medicine for treating hepatocellular carcinoma with abdominal metastasis. Background Art

[0004] Hepatocellular carcinoma (HCC) has consistently ranked among the highest cancer incidence and mortality rates in my country in recent years. The disease presents insidiously, with approximately 80% of patients diagnosed at an advanced stage, precluding the opportunity for curative treatment. While surgical resection is the preferred treatment, only approximately 15% of HCC patients are eligible for such intervention. Approximately 70% of these patients will experience recurrence within 5 years, and if portal vein tumor thrombus (PVTT) or extrahepatic metastasis (EHM) develops, the median survival is only 3-6 months.

[0005] Currently, there are very limited drug therapies for the treatment of HCC. Previous studies have shown that liver cancer stem cells play an important role in the recurrence and metastasis of liver cancer. Systemic treatment options for HCC, such as immunotherapy, targeted therapy or intravenous chemotherapy, have generally had unsatisfactory results and obvious toxic side effects. Sorafenib tosylate (Nexavar from Bayer; a multikinase inhibitor that exerts anti-proliferative effects (RAF1, BRAF and KIT), anti-angiogenic effects (vascular endothelial growth factor receptor [VEGFR] and platelet-derived growth factor receptor" [PDGFRB]) and pro-apoptotic effects) and lenvatinib mesylate (Lenvima from Merck; VEGFR Multikinase inhibitors of VEGF receptors 1-3, fibroblast growth factor receptors (FGFR) 1-4, RET, KIT, and PDGFRa are the only drugs approved for first-line systemic treatment of advanced HCC that cannot be surgically removed. Second-line therapies include multikinase inhibitors such as regorafenib and cabozantinib, anti-VEGFR2 mAb, ramucirumab, and immune checkpoint inhibitors (anti-PD-1 mAb), nivolumab and pembrolizumab. Although immunotherapy and targeted therapy may be effective to some extent, While the drug has demonstrated significant efficacy against HCC, clinically, it faces challenges such as drug resistance and high costs, which patients must contend with during treatment. Furthermore, there are no specific chemotherapy regimens for HCC; instead, generic chemotherapy regimens for digestive system tumors are typically used. These regimens often suffer from poor efficacy and high systemic toxicity. During clinical application, patients are often unable to complete a full cycle of treatment due to the adverse effects of chemotherapy. Therefore, finding a highly effective, low-toxic, inexpensive, and less susceptible to drug resistance treatment regimen or pharmaceutical composition for HCC is an urgent and unmet clinical need.

[0006] Retinoic acid is also known as all-trans-Retinoic acid (ATRA), retinoic acid, retinoic acid, etc. Its chemical name is (13E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohexenyl)-2,4,6,8-nonatetraenoic acid, and its molecular formula is C 20 H 28O2, primarily a derivative of retinoic acid, is an intermediate metabolite of vitamin A in the body and is essential for maintaining growth and development. It has strong cell differentiation-inducing and immunomodulatory effects, making it the preferred drug for treating hematologic malignancies such as acute promyelocytic leukemia (APL) and myelodysplasia in China. It also has promising applications in other clinical areas, including skin diseases, solid tumors, and vascular diseases.

[0007] As a differentiation inducing agent, retinoic acid has a certain effect in clinical use for the treatment of advanced liver cancer. Previous studies have shown that retinoic acid can induce the differentiation of CD133+ liver cancer stem cells, increase the sensitivity of liver cancer cells to cytotoxic drugs, and thus enhance the efficacy of treatment. However, the main problem causing the poor prognosis and survival rate of HCC patients is the abdominal metastasis and distal metastasis caused by HCC, and there has been no research on whether retinoic acid combined with other drugs can significantly reduce the abdominal metastasis of hepatocellular carcinoma. In the process of carrying out clinical research, the inventors unexpectedly found that retinoic acid combined with intravenous FOLFOX4 (oxaliplatin / pentafluorouracil / leucovorin) chemotherapy has a good effect on patients with liver cancer and abdominal metastasis.

[0008] Based on the above findings, the present inventors provide a novel use of a pharmaceutical composition comprising retinoic acid, which is highly effective, low-toxic, inexpensive and not prone to drug resistance, for treating hepatocellular carcinoma with abdominal metastasis. Summary of the Invention

[0009] The purpose of the present invention is to provide a pharmaceutical composition for treating hepatocellular carcinoma with abdominal metastasis that is highly effective, low-toxic, inexpensive, and not prone to drug resistance, comprising retinoic acid, for the treatment of patients with hepatocellular carcinoma with abdominal metastasis, thereby providing a new approach for the safe, effective, convenient, and economical clinical treatment of hepatocellular carcinoma with abdominal metastasis.

[0010] Specifically, the present invention is achieved through the following technical solutions:

[0011] In a first aspect, the present invention provides a pharmaceutical composition for treating hepatocellular carcinoma with abdominal metastasis, the pharmaceutical composition comprising a FOLFOX4 chemotherapy regimen and retinoic acid, wherein the FOLFOX4 chemotherapy regimen is the main active ingredient for treating hepatocellular carcinoma with abdominal metastasis, and the retinoic acid is an auxiliary ingredient for enhancing the FOLFOX4 chemotherapy regimen in treating hepatocellular carcinoma with abdominal metastasis.

[0012] As an optional manner, in the above pharmaceutical composition, the ratio of the FOLFOX4 chemotherapy regimen to the retinoic acid is 1-10:7 by weight.

[0013] As an optional manner, in the above pharmaceutical composition, the ratio of the FOLFOX4 chemotherapy regimen to the retinoic acid is 4-6:7 by weight.

[0014] As an optional manner, in the above pharmaceutical composition, the ratio of the FOLFOX4 chemotherapy regimen to the retinoic acid is 5:7 by weight.

[0015] As an optional manner, in the above pharmaceutical composition, the FOLFOX4 chemotherapy regimen consists of oxaliplatin, 5-fluorouracil and folinate.

[0016] As an optional manner, in the above pharmaceutical composition, the weight ratio is oxaliplatin: 5-fluorouracil: calcium folinate = 1:1-10:1-10.

[0017] As an optional manner, in the above pharmaceutical composition, the weight ratio is oxaliplatin:5-fluorouracil:calcium folinate=1:5-10:1-5.

[0018] As an optional manner, in the above pharmaceutical composition, the weight ratio is oxaliplatin: 5-fluorouracil: calcium folinate = 1:6.5:2.5.

[0019] As an optional mode, in the above-mentioned pharmaceutical composition, the pharmaceutical composition further comprises other drugs having liver-protecting effects.

[0020] Preferably, the other drugs having liver-protecting effects are one or more of Schisandra chinensis, puerarin, silymarin, reduced glutathione, glutamine or vitamin D.

[0021] As an optional approach, in the above pharmaceutical composition, by combining the various active ingredients in the pharmaceutical composition, a significant synergistic effect can be achieved in the treatment of hepatocellular carcinoma with abdominal metastasis.

[0022] In a second aspect, the present invention provides use of the pharmaceutical composition described in the first aspect in preparing a medicament for treating hepatocellular carcinoma with abdominal metastasis.

[0023] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention combines the applicant's advantages in researching and developing retinoic acid to screen out a pharmaceutical composition with synergistic effects in treating liver cancer with abdominal metastasis, significantly enhancing the effect of the FOLFOX4 chemotherapy regimen in treating hepatocellular carcinoma with abdominal metastasis, and significantly increasing the safety of using such drugs and patients' medication compliance. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1: Photos of tumors removed from mice in each group at the end of the experiment in Example 1.

[0027] Figure 2: Average tumor weight of mice in each group at the end of the experiment in Example 1.

[0028] Figure 3: In vivo imaging monitoring of mice in each group during the study and at the end of the experiment in Example 2.

[0029] Figure 4: Imaging findings before and after treatment in a representative patient case of hepatocellular carcinoma with abdominal metastasis. DETAILED DESCRIPTION

[0030] The present inventors, through extensive screening and in-depth research on the mechanism of retinoic acid in treating advanced liver cancer, have discovered for the first time that combining the FOLFOX4 chemotherapy regimen with retinoic acid can significantly enhance the efficacy of the FOLFOX4 chemotherapy regimen in treating hepatocellular carcinoma with abdominal metastasis. Based on this, the present invention was completed.

[0031] As used herein, "liver cancer with abdominal metastasis" refers to advanced hepatocellular carcinoma with abdominal metastasis.

[0032] As used herein, "auxiliary ingredients" generally refer to substances that have no or almost no target pharmacological activity but can enhance the target pharmacological activity of the main active ingredient. The target pharmacological activity of the present invention is mainly the effect of treating hepatocellular carcinoma with abdominal metastasis.

[0033] As used herein, the retinoic acid and FOLFOX4 chemotherapy regimen in the pharmaceutical composition of the present invention are administered in different pharmaceutical formulations. The dosage forms of retinoic acid and FOLOX4 chemotherapy regimen are different, and retinoic acid and FOLOX4 chemotherapy regimen can be administered simultaneously or sequentially.

[0034] In the medical uses described above, the administration time, number of administrations and frequency of administration of "retinoic acid" and "FOLFOX4 chemotherapy regimen" need to be determined according to the specific diagnosis results of the disease, which is within the technical scope mastered by those skilled in the art.

[0035] For example, when a treatment regimen for mice or rats is applied to humans, the effective doses of all drugs for humans can be converted by the effective doses of the drugs for mice or rats, which is also easy to achieve for ordinary technicians in this field.

[0036] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0037] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0038] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0039] Unless otherwise specified, all percentages and parts in the present invention are by weight.

[0040] Example 1: Inhibitory effect of FOLOX4 combined with retinoic acid on the growth of Hub7 subcutaneous transplanted tumors

[0041] 1.1 Experimental Materials and Methods

[0042] In this study, a subcutaneous transplant tumor model was constructed using the liver cancer cell line Huh7 (purchased from ATCC) to detect the inhibitory effect of FOLOX4 combined with retinoic acid on the growth of Hub7 subcutaneous transplant tumors.

[0043] 2×10 6 Huh7 liver cancer cells were injected into the right groin of 3-4 week old NSG mice. 3 The mice were randomly divided into 4 groups: model group, retinoic acid group, FOLOX4 group and combination group, with 5 mice in each group.

[0044] The dosing schedule is as follows:

[0045] (1) Model group: 5% glucose was intraperitoneally injected once a week and normal saline was administered orally once a day;

[0046] (2) Retinoic acid group: 20 mg / kg retinoic acid was administered orally once daily;

[0047] (3) FOLOX4 group: FOLOX4 regimen was administered intraperitoneally once a week, including oxaliplatin 10 mg / kg, 5-Fu 65 mg / kg, and folinic acid 25 mg / kg;

[0048] (4) Combination group: 20 mg / kg of retinoic acid was administered orally once daily, and FOLOX4 regimen was administered intraperitoneally once a week, including 10 mg / kg of oxaliplatin, 65 mg / kg of 5-Fu, and 25 mg / kg of folinic acid.

[0049] The above regimen was administered continuously for 6 weeks.

[0050] The weight of the mice was monitored every three days and the size of the tumor was measured with a caliper. The volume of the tumor was calculated using the formula V = (length × width squared) / 2, where length and width are the maximum longitudinal and transverse diameters, respectively. The trend of tumor changes can be monitored by plotting a tumor volume curve.

[0051] At the end of the experiment after 6 weeks of drug administration, the mice were euthanized and the tumors were removed and photographed.

[0052] The experimental data were statistically analyzed using Graphpad Prism 8.0 software. The experimental data results were expressed as mean ± SD. Data statistics involved t-test or one-way analysis of variance (ANOVA) statistical methods. P < 0.05 indicated that the data differences were statistically significant.

[0053] 1.2 Experimental Results

[0054] Photos of the tumors removed from each group of mice at the end of the experiment are shown in Figure 1. The average tumor weight of each group of mice is shown in Figure 2. The experimental results showed that compared with the model group, both the retinoic acid group and the FOLFOX4 group were able to significantly reduce the tumor weight of the experimental animals, and the FOLFOX4 regimen had a better inhibitory effect on tumors than retinoic acid. When the FOLFOX4 regimen was combined with retinoic acid, the inhibitory effect of the combination group on the tumors of experimental animals became more significant, and the tumor weight of the animals in the combination group was significantly different from that in the FOLFOX4 regimen group (p<0.001). The above results suggest that the combination of the FOLFOX4 regimen and retinoic acid produced a synergistic inhibitory effect on the growth of Hub7 subcutaneous transplanted tumors.

[0055] Example 2: Inhibitory effect of FOLOX4 combined with retinoic acid on Hub7 abdominal metastases

[0056] 2.1 Experimental Materials and Methods

[0057] In this study, Huh7 cells expressing luciferase were used to construct an abdominal metastasis model to observe the inhibitory effect of FOLOX4 combined with retinoic acid on the growth of liver cancer abdominal metastasis. 6 Huh7 liver cancer cells were intraperitoneally injected into NSG mice. On day 14, small animal bioimaging (BLI) demonstrated successful establishment of an intraperitoneal tumor model. The animals were then randomly divided into four groups: a model group, a retinoic acid group, a FOLOX4 group, and a combination group, with five mice in each group.

[0058] The dosing schedule is as follows:

[0059] (1) Model group: 5% glucose was intraperitoneally injected once a week and normal saline was administered orally once a day;

[0060] (2) Retinoic acid group: 20 mg / kg retinoic acid was administered orally once daily;

[0061] (3) FOLOX4 group: FOLOX4 regimen was administered intraperitoneally once a week, including oxaliplatin 10 mg / kg, 5-Fu 65 mg / kg, and folinic acid 25 mg / kg;

[0062] (4) Combination group: 20 mg / kg of retinoic acid was administered orally once daily, and FOLOX4 regimen was administered intraperitoneally once a week, including 10 mg / kg of oxaliplatin, 65 mg / kg of 5-Fu, and 25 mg / kg of folinic acid.

[0063] The above regimen was administered continuously for 7 weeks.

[0064] Mice were monitored by in vivo imaging at regular intervals during the study and at the end of the experiment.

[0065] The experimental data were statistically analyzed using Graphpad Prism 8.0 software. The experimental data results were expressed as mean ± SD. Data statistics involved t-test or one-way analysis of variance (ANOVA) statistical methods. P < 0.05 indicated that the data differences were statistically significant.

[0066] 2.2 Experimental Results

[0067] Figure 3 shows the in vivo imaging monitoring of mice in each group during and at the end of the study. At the end of the study, the fluorescence ROI values ​​for each group are shown in Table 1. The experimental results showed that compared with the model group, the FOLFOX4 regimen alone had a weaker inhibitory effect on Hub7 peritoneal metastases, while retinoic acid alone had a moderate inhibitory effect on Hub7 peritoneal metastases (p < 0.01). However, when FOLFOX4 was combined with retinoic acid, the combination group significantly inhibited Hub7 peritoneal metastases. The experimental results of the combination group were significantly different from those of the model group and the retinoic acid group alone (p < 0.001 and p < 0.001, respectively). These results suggest that the combination of FOLFOX4 and retinoic acid produces a synergistic inhibitory effect on the growth of Hub7 peritoneal metastases.

[0068] Table 1: Fluorescence ROI values ​​of mice in each group at the end of the study (mean ± SD)

[0069] Note: Compared with the model group, ** p<0.01, *** p<0.001; compared with the retinoic acid group,## p<0.01.

[0070] Example 3: Representative Case of Hepatocellular Carcinoma with Peritoneal Metastasis

[0071] A 67-year-old male patient, Mr. Ni (from the Department of Liver Surgery, Shanghai Oriental Hepatobiliary Surgery Hospital, a co-development partner for the clinical trial), was found to have multiple abdominal metastases after liver cancer surgery. He was treated with six cycles of ATRA+FOLFOX4, and the abdominal metastases disappeared. Figure 1 shows the pre- and post-treatment imaging results of a representative patient with hepatocellular carcinoma and abdominal metastases.

[0072] The combined treatment regimen is as follows:

[0073] Chemotherapy usage:

[0074] Oxaliplatin 85 mg / m 2 (Day 1), 200 mg / m2 of calcium folinate 2 (Day 1), 5-FU 400 mg / m 2 (Day 1), 5-FU 600 mg / m 2 (Last 40 hours).

[0075] Retinoic acid usage:

[0076] 20 mg orally 3 times a day, starting 3 days before chemotherapy and continuing for 5 days.

[0077] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A pharmaceutical composition for treating hepatocellular carcinoma with abdominal metastasis, characterized in that: The pharmaceutical composition comprises FOLFOX4 chemotherapy regimen and retinoic acid, wherein the FOLFOX4 chemotherapy regimen is the main active ingredient for treating hepatocellular carcinoma with abdominal metastasis, and the retinoic acid is an auxiliary ingredient for enhancing the FOLFOX4 chemotherapy regimen for treating hepatocellular carcinoma with abdominal metastasis.

2. The pharmaceutical composition according to claim 1, characterized in that: In terms of weight ratio, the dosage ratio of the FOLFOX4 chemotherapy regimen to the retinoic acid is 1-10:

7.

3. The pharmaceutical composition according to claim 2, characterized in that: In terms of weight ratio, the dosage ratio of the FOLFOX4 chemotherapy regimen to the retinoic acid is 4-6:

7.

4. The pharmaceutical composition according to claim 1, characterized in that: The FOLFOX4 chemotherapy regimen consists of oxaliplatin, 5-fluorouracil and folinate.

5. The pharmaceutical composition according to claim 4, characterized in that: In terms of weight ratio, oxaliplatin: 5-fluorouracil: calcium folinate = 1: 1-10: 1-10.

6. The pharmaceutical composition according to claim 5, characterized in that: In terms of weight ratio, oxaliplatin: 5-fluorouracil: calcium folinate = 1:5-10:1-5.

7. The pharmaceutical composition according to claim 1, characterized in that: The pharmaceutical composition also contains other drugs with liver protection effects.

8. The pharmaceutical composition according to claim 7, characterized in that: The other drugs with liver protection effect are one or more of Schisandra chinensis, puerarin, silymarin, reduced glutathione, glutamine or vitamin D.

9. The pharmaceutical composition according to any one of claims 1 to 8, characterized in that: The combination of various active ingredients in the pharmaceutical composition has a significant synergistic effect in treating hepatocellular carcinoma with abdominal metastasis.

10. Use of the pharmaceutical composition according to any one of claims 1 to 9 in the preparation of a medicament for treating hepatocellular carcinoma with abdominal metastasis.

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