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CN-121987790-A - Hepatocellular carcinoma drug resistance reversal product targeting ABHD6 mediated drug resistance pathway and application thereof

CN121987790ACN 121987790 ACN121987790 ACN 121987790ACN-121987790-A

Abstract

The application provides a hepatocellular carcinoma drug resistance reversal product targeting an ABHD 6-mediated drug resistance pathway and application thereof, relates to the technical field of biomedicine, and discloses application of the ABHD 6-mediated drug resistance pathway serving as a target in preparation of a drug resistance reversal product for hepatocellular carcinoma (HCC) and lenvatinib and related biomarkers. The pathway core mechanism is lactic acid to induce the K245 site of ABHD6 to be lactated, promote the mitochondrial translocation and competitive combination with FIS1, lead to excessive mitochondrial fusion, and inhibit apoptosis and ROS generation. The product comprises an ABHD 6K 245 lactate inhibitor, an ABHD6-FIS1 interaction blocker, an allosteric inhibitor WWL70, an LDHA inhibitor and a natural substrate 2-AG/C16:0 MAG. Cell, xenograft, PDX and in situ liver transplantation model verification shows that the products can effectively reverse drug resistance, and the ABHD 6K 245 lactate level can predict drug resistance. The application fills the blank of a drug resistance mechanism of the lenvatinib, provides a plurality of high-efficiency drug resistance reversal products and accurate prediction markers, and provides a new scheme for clinical treatment.

Inventors

  • TANG ZHIYUAN
  • SUN YUENING
  • Luo Chengju

Assignees

  • 南通大学附属医院

Dates

Publication Date
20260508
Application Date
20260120

Claims (9)

  1. The application of the ABHD6 mediated drug resistance pathway serving as a target point in preparing a product for reversing the drug resistance of hepatocellular carcinoma (HCC) and lenvatinib is characterized in that the sequence of the ABHD6 is shown as SEQ ID NO. 01.
  2. 2. The use of the ABHD 6-mediated drug resistance pathway as a target in the preparation of a product for reversing the drug resistance of hepatocellular carcinoma (HCC) and lenvatinib according to claim 1, wherein the ABHD 6-mediated drug resistance pathway induces lactic acid at the K245 site of ABHD6, promotes the translocation of the mitochondria of ABHD6 and competitive binding with mitotic protein 1 (FIS 1), and replaces the dynamics related protein (DRP 1) to cause excessive mitochondrial fusion, thereby inhibiting apoptosis and Reactive Oxygen Species (ROS) production.
  3. 3. The use of the ABHD6 mediated drug resistant pathway of claim 1 as a target for the preparation of a product for reversing the resistance of hepatocellular carcinoma (HCC) and lenvatinib, wherein said product is selected from at least one of the group consisting of an agent that inhibits lactate at the ABHD 6K 245 site, an agent that blocks the interaction of ABHD6 with FIS1, an allosteric inhibitor that binds to the catalytic site of ABHD 6S 148, an agent that inhibits lactate production, and a natural substrate for ABHD 6.
  4. 4. The use of the ABHD 6-mediated drug resistance pathway as a target in the preparation of a product for reversing the resistance of hepatocellular carcinoma (HCC) and lenvatinib, wherein the allosteric inhibitor comprises WWL70.
  5. 5. The use of the ABHD 6-mediated drug resistance pathway as a target in the preparation of a product for reversing the resistance of hepatocellular carcinoma (HCC) and lenvatinib, wherein the agent for inhibiting the production of lactic acid comprises a lactate dehydrogenase A chain (LDHA) inhibitor.
  6. 6. The use of the ABHD 6-mediated drug resistance pathway of claim 3 as a target for the preparation of a product for reversing the resistance of hepatocellular carcinoma (HCC) and lenvatinib, wherein the natural substrate is 2-arachidonylglycerol (2-AG) or C16:0 monoacylglycerol (C16:0 MAG).
  7. 7. A product for reversing the drug resistance of hepatocellular carcinoma lenvatinib is characterized in that the product takes an ABHD6 mediated drug resistance pathway as a target point.
  8. 8. A product for reversing the resistance of hepatocellular carcinoma, as set forth in claim 7, characterized in that it further comprises other auxiliary agents acceptable in medicine.
  9. Application of K245 site lactate level of ABHD6 in preparing drug resistance prediction biomarker of lenvatinib.

Description

Hepatocellular carcinoma drug resistance reversal product targeting ABHD6 mediated drug resistance pathway and application thereof Technical Field The application relates to the technical field of biomedicine, in particular to a hepatocellular carcinoma drug resistance reversal product targeting an ABHD6 mediated drug resistance pathway and application thereof. Background Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide, accounting for over 50% of new cases and deaths annually in china. While Tyrosine Kinase Inhibitors (TKIs) such as lenvatinib improve survival outcomes in patients with advanced HCC, about 50% of patients develop resistance within 6-7 months of treatment, with median survival only slightly exceeding 12 months due to treatment failure. This resistance stems from heterogeneous mechanisms involving metabolic reprogramming and mitochondrial adaptation, however current strategies for kinase activity have limited efficacy on drug-resistant persisting cells (drug tolerant persisters). Disclosure of Invention The application aims to solve the technical problem that the strategy for kinase activity in the prior art has limited curative effect on the drug-resistant persistent cell. In order to solve the technical problems, the application provides application of an ABHD6 mediated drug resistance pathway serving as a target point in preparation of a product for reversing drug resistance of hepatocellular carcinoma (HCC) and lenvatinib, wherein the sequence of the ABHD6 is shown as SEQ ID NO: 01. Compared with the prior art, the application at least comprises the following beneficial effects: 1. The application firstly clarifies the ABHD6 mediated lactic acid dependent drug resistance path, fills the blank of HCC (drug resistance) drug resistance mechanism research, and provides a clear target for drug resistance intervention; 2. The application provides a drug resistance reversal product with different structures and action mechanisms, has flexible application scene, can be used singly or in combination with lenvatinib, and meets different clinical requirements; 3. the application discovers and verifies the specific biomarker of ABHD 6K 245 site lactate, can predict the sensitivity of a patient to lenvatinib in advance, avoids invalid treatment, and reduces medical cost; 4. The application clearly shows that the ABHD6 mediated drug resistance is independent of the catalytic activity, provides a theoretical basis for developing a targeting drug which does not influence the normal physiological function of the ABHD6, and reduces potential side effects. Drawings In the present application, the data of the drawings are expressed as mean ± SD, and the statistical significance is defined as P <0.05, P <0.01, P <0.001, and "ns" indicates no significant difference, and unless otherwise indicated, the comparison between the groups uses a one-way variance analysis, and the comparison between the two groups uses a t-test. FIG. 1 single cell transcriptome identification of ABHD6 as a key driver for HCC drug resistance to lenvatinib. A is Lenva-R volcanic plot of non-drug resistant hepatocytes DEGs, ABHD6 is the most significantly up-regulated metabolic gene, and B is IHC validation of ABHD6 protein elevation in Lenva-R recurrent HCC tumor (n=20, scale bar 100 μm). FIG. 2 ABHD6 drives HCC cell resistance to lenvatinib. A is CCK-8 experiment showing that the ABHD6 operation bidirectionally regulates the sensitivity of Huh7 cells to lenvatinib (n=10), B is Lenva-R HepG2 and Huh7 cell lines and 20 mu M of lenvatinib, the influence of the ABHD6KO on cell proliferation, C is NOD-SCID mice in vivo tumor growth fluorescence imaging (n=5/group), the curative effect of the ABHD6-KO on Lenva-R xenograft is recovered, D is the tumor weight quantification (n=5) of the experimental end point (day 28) of the HepG2 cell xenograft model, E, F is Huh7 cell apoptosis flow analysis (n=5), E shows that the ABHD6-OE inhibits apoptosis induced by the lenvatinib, and F shows that the apoptosis of the ABHD6-KO is enhanced Lenva-R cells. Figure 3 xenograft tumor model the effect of ABHD6 (S148A) on the sensitivity of lenvatinib (n=5) was evaluated. Figure 4ABHD6 promotes drug resistance by modulating mitochondrial dynamics. A is Co-IP and mass spectrum identification FIS1 is ABHD6 direct interaction factor, B is mito-EGFP marked mitochondrial confocal image, lenva-R HCC cell mitochondrial network extension and over fusion (scale 10 μm, n=3, t test), C is Seahorse mitochondrial pressure test to show the effect of ABHD6 knockout on Lenva-R cell mitochondrial respiratory function (n=5), D, E is mitochondrial dynamics related western blotting and relative quantification (n=3, t test), F is FIS1 KD/OE verification and CCK-8 test to evaluate the effect on ABHD6 mediated drug resistance. Figure 5ABHD6 maintains mitochondrial homeostasis and inhibits mitochondrial autophagy. A is DCF-DA staining flow ass