CN114681609A

CN114681609A - Application of anti-IL-6 antibody composition in preparation of drugs for treating hepatocellular carcinoma

Info

Publication number: CN114681609A
Application number: CN202210481644.1A
Authority: CN
Inventors: 龚渭华; 苗小龙; 王垚; 姜源聪
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2022-05-05
Filing date: 2022-05-05
Publication date: 2022-07-01

Abstract

The invention provides application of an anti-IL-6 antibody composition in preparing a medicine for treating hepatocellular carcinoma, wherein the composition is prepared by combining an anti-IL-6 antibody and a phosphoinositide-3-kinase-rapamycin target small molecule inhibitor (NVP-BEZ 235). Based on the discovery that the expression level of proinflammatory cytokine IL-6 is increased after NVP-BEZ235 is taken, experiments prove that the composition group remarkably delays the tumor process and prolongs the survival time of tumor-bearing mice after treatment. Furthermore, the combination of the invention is expected to prepare a medicine which can improve the treatment effect of NVP-BEZ235 and can be applied to the treatment of hepatocellular carcinoma.

Description

Application of anti-IL-6 antibody composition in preparation of drugs for treating hepatocellular carcinoma

Technical Field

The invention belongs to the field of biomedicine, relates to an application of an anti-IL-6 antibody composition in preparation of a medicine for treating hepatocellular carcinoma, and relates to an application of an anti-IL-6 antibody and NVP-BEZ235 (phosphoinositide-3-kinase-rapamycin target spot small molecule inhibitor) composition in preparation of a medicine for treating hepatocellular carcinoma.

Background

Primary liver cancer occupies the prostate, the most cancer-related death, particularly Hepatocellular carcinoma (HCC), accounting for almost 90% of all cases of primary liver cancer. The liver cancer treatment is limited in selection and high in invasion degree, local treatment modes such as radio frequency ablation, hepatectomy and liver transplantation are usually selected for patients with liver cancer in early and middle stages, and general treatment is usually adopted for patients with liver cancer in late stage, and the use of multi-kinase inhibitors sorafenib and ranvatinib is the current main first-line treatment. Although the conventional therapy has a certain antitumor effect, the prognosis effect cannot be improved remarkably.

Most drugs have not shown significant survival benefit over the last 10 years. Currently, new therapeutic approaches to hepatocellular carcinoma involve strategies for combination therapy with molecular targeted inhibitors to achieve better synergy by enhancing the sensitivity of targeted drugs. Therefore, the development of new treatment strategies provides a certain reference value for the treatment of hepatocellular carcinoma.

The phosphoinositide-3-kinase-rapamycin target inhibitor Dactlisib (NVP-BEZ235) is a dual ATP competitive PI3K and mTOR inhibitor and is currently in preclinical phase I/II trials. Drug toxicity is a common challenge for small molecule inhibitors, and many patients treated with PI3K inhibitors have adverse reactions such as pneumonia, colitis, immune-related toxicity, etc. in previous clinical reports.

In previous experiments, the single administration of the compound can play an anti-tumor effect by inhibiting tumor proliferation, and the NVP-BEZ235 is found to cause the level of proinflammatory cytokine IL-6 to be increased after the administration of the compound. The occurrence of inflammatory responses is closely related to the development of tumors, and cytokines are considered as key mediators connecting the two. IL-6 has been shown to promote the development of a variety of tumor cells, while a number of clinical samples show that the IL-6 content in the serum of liver cancer patients is significantly higher than that of healthy patients, with a poorer prognosis. In response to this finding, we propose: a regimen for targeted inhibition of IL-6 while treating HCC in combination with NVP-BEZ 235. For the research of molecular targeted drug combination therapy, the defect of single drug use is made up to a certain extent, the drug toxicity is avoided, and the aim of enhancing the NVP-BEZ235 anti-tumor effect is achieved.

Disclosure of Invention

The invention aims to provide an application of an anti-IL-6 antibody composition in preparing a medicine for treating hepatocellular carcinoma, in particular to an application of an anti-IL-6 antibody and NVP-BEZ235 (phosphoinositide-3-kinase-rapamycin target small molecule inhibitor) composition in preparing a medicine for treating hepatocellular carcinoma, wherein the composition is prepared by combining the anti-IL-6 antibody and the NVP-BEZ235 according to the combination ratio of 10 mg/kg: 60 mg/kg. The medicine is prepared from the composition and pharmaceutic adjuvants.

According to the invention, a mouse hepatocellular carcinoma model is constructed by using a hydrodynamic transfection method, the IL-6 expression is found to be increased after NVP-BEZ235 is taken (figure 1), and a medication scheme is designed according to the tumor process (figure 2). Experiments prove that the liver weight/body weight, the maximum tumor diameter and the tumor node number of mice in an observation group treated by the composition are obviously reduced after treatment (figure 3), and the survival time is obviously prolonged (figure 4). Treatment with the composition group significantly reduced tumor burden (figure 5). The Ki-67 positive rate of the liver cancer region of the composition group is obviously reduced (figure 6). The composition of the invention can reduce the side effect of NVP-BEZ235 medication and simultaneously achieve the aim of synergistic treatment of hepatocellular carcinoma. The medicine prepared from the composition can improve the treatment effect of NVP-BEZ235, and is applied to treatment of hepatocellular carcinoma.

Drawings

FIG. 1 shows H & E detection of IL-6 protein expression in HCC tissues of mice from different treatment groups.

FIG. 2 shows the construction and drug treatment process of HCC in mice of different treatment groups.

FIG. 3 is a graph showing the results of liver weight/body weight, maximum tumor diameter, and tumor nodule number of tumor-bearing mice.

FIG. 4 is a statistical plot of tumor burden survival time in mice.

FIG. 5 shows gross appearance and H & E staining of liver specimens from mice in different treatment groups.

FIG. 6 shows the expression of tumor-bearing Ki-67 in immunohistochemical detection mice.

Detailed Description

The invention is further explained by the accompanying drawings and examples.

Example 1

The influence of the anti-IL-6 antibody and NVP-BEZ235 composition and the tumor bearing index on the tumor bearing survival time is detected by constructing a mouse hepatocellular carcinoma model.

1. Construction of mouse hepatocellular carcinoma model

1.1 experimental animals and main materials: male inbred C57 mice, weighing 20-25g, were purchased from Shanghai Spideco.

1.2 method: wild type C57 mouse is selected, and a plasmid-induced mouse hepatocellular carcinoma (HCC) model is constructed. According to the C-myc plasmid: n-ras plasmid: the plasmid SB was added to sterile 1 × PBS at a ratio of 1:19:2 to prepare a plasmid mixture at a concentration of 11 μ g/mL; weighing 8-10 week old mice, and extracting the corresponding volume (mL) of the mixed plasmid solution according to 10% of the weight (g) of the mice; fixing the mouse in a venous visual mouse tail injection fixer, pressing the tail of the mouse until the tail vein is fully exposed, and injecting the mixed plasmid into the mouse body by using an injector within 5-9 seconds; and (5) after the state of the mouse is recovered to be normal, the mouse is returned to the cage for continuous feeding.

2. And (5) experimental results. Firstly, a mouse hepatocellular carcinoma model is constructed, and compared with a control group of mice, the NVP-BEZ235 after being taken can cause the expression level of proinflammatory cytokine IL-6 to be increased (figure 1). After different drug treatments (FIG. 2), the NVP-BEZ 235-treated group and the IL-6 antibody-treated group were able to inhibit the tumor progression of HCC to some extent but were very limited. The composition-treated mice showed a significant reduction in the indices of liver-to-weight ratio, maximum tumor diameter and tumor nodule number compared to the control mice (fig. 3). Survival analysis showed that the tumor-bearing survival time was most significantly prolonged in the mice of the composition group (fig. 4). The combination treatment significantly delayed tumor progression as shown by gross appearance of liver specimens and H & E staining (figure 5). The tumor area Ki-67 was significantly reduced in the composition group mice (FIG. 6).

Claims

1. The application of the anti-IL-6 antibody composition in the preparation of the drugs for treating hepatocellular carcinoma is characterized in that the composition consists of the anti-IL-6 antibody and phosphoinositide-3-kinase-rapamycin target small molecule inhibitors.

2. Use according to claim 1, wherein the composition is in a combined ratio of: anti-IL-6 antibodies: the phosphoinositide-3-kinase-rapamycin target small molecule inhibitor is 10 mg/kg: 60 mg/kg.

3. The use of claim 1, wherein the medicament is prepared from the composition and a pharmaceutically acceptable excipient.