CN110227159B - Medicine for improving drug resistance of solid tumor to anti-vascular drugs and application of CPT1a inhibitor in medicine - Google Patents
Medicine for improving drug resistance of solid tumor to anti-vascular drugs and application of CPT1a inhibitor in medicine Download PDFInfo
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Abstract
The invention provides a new application of a CPT1a inhibitor, in particular to an application in preparing a medicament for improving the drug resistance of solid tumors to anti-vascular drugs. The CPT1a inhibitor acts on CPT1a on the surface of the mitochondrial outer membrane of the tumor cell, inhibits fatty acid from entering a mitochondrial matrix for oxidation, inhibits the uptake of free fatty acid by the tumor cell, restores the sensitivity of the tumor cell to the antiangiogenic drug therapy, inhibits the proliferation of the tumor cell, promotes the apoptosis of the tumor cell, and further can achieve the effect of improving the drug resistance of solid tumor to the antiangiogenic drug. The invention also provides a medicine for improving the drug resistance of the solid tumor to the anti-vascular drug and a medicine for treating the solid tumor.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a medicine for improving the drug resistance of solid tumors to anti-vascular drugs and application of a CPT1a inhibitor in the medicine.
Background
Malignant tumors (such as liver cancer, colon cancer, breast cancer and other solid tumors) are extremely aggressive and threaten human life, and the incidence rate of the malignant tumors is in an increasing trend. The growth rate of malignant tumor cells is high, metabolites are accumulated, and tumor blood vessel disorders are caused. However, the major obstacle faced in anti-vascular therapy is that patients with malignant tumors often develop resistance to anti-vascular drugs, the mechanism of which is largely unknown.
Therefore, there is a need to provide a drug that can improve the resistance of anti-angiogenic drugs in the treatment of solid tumors.
Disclosure of Invention
In view of the above, the invention provides an application of a CPT1a inhibitor in preparing a drug for improving drug resistance of a tumor to an anti-vascular drug and a drug for improving drug resistance of an anti-vascular drug, which can effectively improve drug resistance of an anti-vascular drug in treatment of solid tumors and optimize the existing treatment effect of solid tumors.
In a first aspect, the invention provides the use of an inhibitor of CPT1a in the manufacture of a medicament for the amelioration of resistance to anti-angiogenic drugs in solid tumours.
The CPT1a inhibitor acts on CPT1a (carnitine palmitoyl transferase 1a) on the surface of the mitochondrial outer membrane of the tumor cell, inhibits fatty acid from entering a mitochondrial matrix to be oxidized, inhibits the uptake of free fatty acid by the tumor cell, restores the sensitivity of the tumor cell to an anti-vascular drug therapy, inhibits the proliferation of the tumor cell, reduces the volume of the tumor and promotes the apoptosis of the tumor cell. Thereby improving the drug resistance of the solid tumor to the anti-vascular drug and enhancing the anti-tumor effect of the anti-vascular drug.
During the oxidation of fatty acids beta, long chain fatty acids need to be converted to acylcarnitines to enter the mitochondrial matrix for oxidation. Carnitine palmitoyl transferase 1 (CPT 1), which is the rate-limiting enzyme for fatty acid oxidation, is located in the outer mitochondrial membrane and is overexpressed in many human tumors. CPT1 promotes the transfer of long-chain fatty acids from acyl-CoA onto carnitine from the cytosol into the mitochondria for beta oxidation catalyzed by CPT2 on the inner membrane of mitochondria. CPT1a (carnitine palmitoyl transferase 1a) is a subtype of carnitine palmitoyl transferase 1 (CPT 1).
Wherein the solid tumor includes one or more of liver cancer, lung cancer, pancreatic cancer, kidney cancer, stomach cancer, esophageal cancer, colon cancer, bladder cancer, breast cancer, ovarian cancer and cervical cancer, but is not limited thereto.
Optionally, the CPT1a inhibitor comprises one or more of a chemical drug, a polypeptide drug, a protein drug, and a gene drug that inhibits CPT1 a.
Further, the chemical drugs that inhibit CPT1a inhibitors are generally referred to as small molecule organic compounds. The amino acid drugs for inhibiting CPT1a include but are not limited to natural proteins, recombinant proteins and polypeptides. The "gene-like drug" for inhibiting CPT1a includes but is not limited to nucleic acid fragments, such as DNA fragments and RNA fragments.
In one embodiment of the present invention, the chemical inhibiting CPT1a may be etomoxir.
In another embodiment of the invention, the protein drug inhibiting CPT1a can be a polypeptide against CPT1 a.
In another embodiment of the invention, the nucleic acid segment for inhibiting CPT1a can be CPT1a siRNA drugs for silencing expression of CPT1a, and can also be shRNA plasmids or shRNA lentiviral particles with slow protein conversion of CPT1a target genes.
Optionally, the medicament for improving the resistance of the solid tumor to the anti-vascular drug further comprises a pharmaceutically acceptable carrier. In this case, the medicament contains the CPT1a inhibitor and a pharmaceutically acceptable carrier.
Optionally, the pharmaceutically acceptable carrier includes at least one of a solvent, a polymer, a liposome, a recombinant viral vector, and a eukaryotic recombinant expression vector, but is not limited thereto.
Wherein the solvent includes, but is not limited to, water, physiological saline, and other non-aqueous solvents. The recombinant viral vector may include one or more of a lentiviral vector, an adenoviral vector, and a retroviral vector, but is not limited thereto.
Wherein the polymer comprises one or more of polylysine, polyethyleneimine (branched and/or chain) and modified substances thereof, polyamidoamine dendrimer (PAMAM) and derivatives thereof, polypropyleneimine dendrimer (PPI) and derivatives thereof, chitosan, polylactic-co-glycolic acid (PLGA), polylactic acid, gelatin, cyclodextrin, sodium alginate, albumin and hemoglobin, but is not limited thereto. Among them, polyethyleneimine and its modified products, PAMAM and its derivatives, PPI and its derivatives, chitosan, etc. may be referred to as cationic polymers.
The liposome can be prepared by self-assembly of cationic lipid, neutral auxiliary lipid, cholesterol, and phospholipid (such as soybean lecithin, yolk lecithin, cephalin, etc.), or can be prepared by inserting distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG) into phospholipid layer formed by phospholipid molecules.
In the present invention, the "pharmaceutically acceptable carrier" is used to transport the drug of the present invention to exert its intended effect. In general, delivery is from one organ or portion to another, and the carrier must be compatible with the pharmaceutical composition, not interfere with the biological activity of the drug, and be relatively non-toxic, e.g., the carrier enters the body without causing toxic side effects or having a severe reaction with the drug it carries, which does not adversely affect the patient.
The CPT1a inhibitor as the active ingredient can be dispersed or adsorbed in the carrier to form a dispersion system, or can be coated/encapsulated with the liposome, polymer, etc. to form a spherical structure (e.g., nanocapsule or microcapsule). For example, when the CPT1a inhibitor is a nucleic acid fragment that inhibits CPT1a, the drug that improves the resistance of solid tumors to anti-vascular drugs includes, but is not limited to, cationic polymers, polypeptides, protein drugs, etc. that encapsulate, bind to, or blend with the nucleic acid fragment.
The CPT1a inhibitor encapsulated in the spherical structure can perform sustained release, controlled release or targeted release, so that the drug can exert the optimal efficacy, and the stability of the drug can be improved. For example, albumin, gelatin, chitosan, polylactic acid may generally form microspheres that can disperse or encapsulate a pharmaceutically active ingredient.
Optionally, in the drug for improving the resistance of the solid tumor to the anti-vascular drug, the content of the CPT1a inhibitor is 0.1-10 mg/kg.
Optionally, the CPT1a inhibitor as a single active ingredient or with other pharmaceutically acceptable active ingredients constitutes the active ingredient in the medicament for improving the resistance of solid tumors to anti-angiogenic drugs.
Optionally, the medicament for improving the resistance of the solid tumor to the anti-vascular drug further comprises one or more of a diluent and an excipient.
The primary function of the diluent is to fill the weight or volume of the tablet to facilitate tableting. Optionally, the diluent comprises one or more of starches, sugars, celluloses, and inorganic salts. The excipient is additive except main active ingredients in the medicine, and comprises binder, filler, disintegrating agent, lubricant, wine, vinegar, medicinal juice, etc. in the tablet, matrix part in semi-solid preparation ointment and cream, antiseptic, antioxidant, corrective, aromatic, cosolvent, emulsifier, solubilizer, osmotic pressure regulator, colorant, etc. in the liquid preparation.
Optionally, the form of the drug for improving the resistance of the solid tumor to the anti-vascular drug comprises a tablet, a capsule, a powder, a granule, a pill, a syrup, a solution or a suspension. The form of the drug for improving the resistance of solid tumors to anti-vascular drugs depends on the practical use.
Alternatively, the drug for improving the resistance of the solid tumor to the anti-vascular drug is administered orally or by injection.
Preferably, the drug for improving the resistance of the solid tumor to the anti-vascular drug is administered by injection. In this case, the drug for improving the resistance of the solid tumor to the anti-vascular drug is preferably in the form of a solution, for example, dissolved in water or physiological saline. Further, the injection is administered by intraperitoneal injection, subcutaneous injection, intramuscular injection or intravenous injection.
Alternatively, the drug for improving the resistance of a solid tumor to an anti-vascular drug may be administered locally or systemically.
Optionally, the amount of the drug for improving the resistance of the solid tumor to the anti-vascular drug is 5 to 20 mg/kg of body weight per day. In particular, the amount of the drug that improves the resistance of a solid tumor to an anti-vascular drug depends on a variety of factors including, but not limited to, the desired biological activity and the tolerance of the subject to the drug.
The CPT1a inhibitor can inhibit fatty acid from entering a mitochondrial matrix of tumor cells to be oxidized, inhibit the uptake of free fatty acid by the tumor cells, restore the sensitivity of the tumor cells to the anti-vascular drug therapy, inhibit the proliferation of the tumor cells, promote the apoptosis of the tumor cells, further improve the effect of the solid tumors on the anti-vascular drug resistance and improve the treatment effect on malignant solid tumors. The new application of the CPT1a inhibitor provided by the invention opens up a new way for treating solid tumors and has a more obvious treatment effect.
In a second aspect, the present invention provides a drug for improving the resistance of a solid tumor to an anti-vascular drug, wherein the drug for improving the resistance of the solid tumor to the anti-vascular drug comprises a CPT1a inhibitor and a pharmaceutically acceptable carrier.
The effect of the CPT1a inhibitor, the pharmaceutically acceptable carrier, and the like are as described in the first aspect of the invention and will not be described herein.
The invention provides a medicament for improving the resistance of solid tumors to anti-vascular drugs, which comprises a CPT1a inhibitor and a pharmaceutically acceptable carrier thereof. The CPT1a inhibitor in the medicine can inhibit fatty acid from entering mitochondrial matrix for oxidation, inhibit free fatty acid uptake of tumor cells, restore sensitivity of tumor cells to antiangiogenic drug therapy, inhibit tumor cell proliferation, promote tumor cell apoptosis, improve the effect of solid tumor on antiangiogenic drug resistance, and improve the treatment effect on malignant solid tumor.
In a third aspect, the invention also provides a medicament for treating solid tumors, which comprises a CPT1a inhibitor and an anti-vascular medicament.
Wherein the mass ratio of the CPT1a inhibitor to the antiangiogenic drug is (1-100): 1.
further, the content of the CPT1a inhibitor is 0.1-10 mg/kg.
Further, the content of the anti-vascular drug is 0.1-10 mg/kg.
Wherein, the anti-vascular drugs include but are not limited to bevacizumab, sorafenib and sunitinib.
Wherein, the anti-vascular medicine is used for reducing the density of tumor blood vessels and causing the tumor cells to lack oxygen; the CPT1a inhibitor can inhibit fatty acid from entering into mitochondrial matrix for oxidation, inhibit free fatty acid uptake of tumor cells in anoxic environment, recover sensitivity of tumor cells to antiangiogenic drug therapy, inhibit tumor cell proliferation, reduce tumor volume, and promote tumor cell apoptosis. Thereby achieving the effect of improving the drug resistance of the solid tumor to the anti-vascular drug. Under the combined action of the CPT1a inhibitor and the antiangiogenic drug, a better antitumor effect is achieved.
Wherein, the medicine for treating the solid tumor also comprises a pharmaceutically acceptable carrier.
Advantages of embodiments of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of embodiments of the invention.
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FIG. 1 is a study of the resistance of various solid tumors to anti-angiogenic drugs in comparative example 1;
FIG. 2 is a graph showing the improvement effect of CPT1a on the resistance to an anti-vascular drug in a colon cancer (CRC) tumor and a liver in situ tumor (HCC) model in a fatty liver mouse in example 1;
FIG. 3 shows the cell growth (a) and cell metabolism (b) of colon cancer cells by the combination of the anti-vascular drug and the drug carrier of CPT1a inhibitor in example 2;
FIG. 4 is a graph showing the improvement of the resistance of CPT1a inhibitor against angiodrug in the colon cancer (CRC) liver metastatic tumor model of fatty liver mice in example 2.
Detailed Description
While the following is a description of the preferred embodiments of the present invention, it should be noted that those skilled in the art can make various modifications and improvements without departing from the principle of the embodiments of the present invention, and such modifications and improvements are considered to be within the scope of the embodiments of the present invention.
The experimental conditions of the invention are introduced:
(1) cell culture
The murine MC38 colon cancer cell line employed in the present invention was provided friendly by Dr. Spezilian, university of Pamplona Nara, Spanish, applied to the department of Gene therapy of the medical research center. The mouse PancO2 pancreatic ductal adenocarcinoma cell line used in the present invention was offered by doctor maxilian Schnurr, university of munich, germany. And the liver cancer cell line adopts Hepa1-6 liver cancer cells.
The cells were cultured in 10% FBS-DMEM (catalog No. SH30243.01, HyClone) medium containing 100U/mL penicillin, 100. mu.g/mL streptomycin (catalog No. SV30010, HyClone), and 2mM L-glutamine (catalog No. SH30034.01, HyClone), respectively. In both the orthotopic liver tumor model and the colon cancer liver metastasis model, tumor cells were stably transfected with luciferase-expressing lentiviral vectors.
(2) Cell proliferation assay and metabolic assay:
MC38 cells were seeded at a density of 2500 cells/well in 96-well plates and incubated in 100. mu.L of the above medium for 24 hours. The cell proliferation was assessed at various time points using the MTT assay system (Cat. No. M5655; Sigma-Aldrich). mu.L of MTT at a concentration of 5. mu.g/L was added to each well of the cell culture, followed by incubation at room temperature for 4 hours. After removing the supernatant, 200. mu.L of 100% DMSO was added to each well, and the cells were suspended by pipetting. The absorbance was measured at 535nm using a microplate reader (BMG LABTECH, Germany).
Metabolic assay of tumor cells Using Hippocampus cell Metabolic Analyzer, a monolayer of CRC tumor cells was seeded on cell culture plates without CO at 37 deg.C2Culturing for 1h under the condition. Oxygen Consumption Rate (OCR) of the substrate each set of metabolic states was measured using an XF24 analyzer.
(3) Animal experiments: the animal experiments of the present application used 6-8 week old male or female C57Bl/6 mice, which were maintained under standard animal feeding conditions and fed either a standard diet or a high fat diet. Wherein, in the fatty liver model, a high-fat diet for 3 months or more is used to induce hepatic steatosis.
(4) Establishing different mouse tumor models:
when different mouse tumor models are constructed, the same tumor cell can be injected into different parts of a healthy mouse; or injecting different tumor cells into a healthy mouse fed with a standard diet or a fatty liver mouse fed with a high fat diet. When investigating the therapeutic effect of different pharmaceutical preparations on mice of different tumor models, the pharmaceutical preparations can be injected intraperitoneally into mice of different experimental groups, and after a period of time, the mice are administered with a lethal dose of CO2Sacrifice, dissect and perform histological examination, RNA and protein extraction, hypoxia, apoptosis, etc.
Comparative example 1: multiple solid tumors in the adipose environment can develop resistance to anti-angiogenic drugs
Subcutaneous transplantation of tumors: the above 1X 10 suspension in 30. mu. L, PBS6Individual MC38 or Panc02 tumor cells were implanted subcutaneously into C57Bl/6 mice (implantation incisions were created in the skin layer and in the inguinal white fat (WAT) region of the non-adipose environment, respectively, for subsequent observation of tumor growth in both the adipose and non-adipose environments). Tumor size was measured with calipers every other day and tumor volume was calculated for subcutaneously transplanted colon cancer (CRC) or PDAC (pancreatic cancer) according to standard formula.
In addition, in establishing the colon cancer (CRC) tumor model of comparative example 1, MC38 cells can be implanted from the spleen to establishLeft subcostal surgical incision, 50. mu.L of 1X 10 in PBS6Individual MC38 cells were injected into the exposed spleen of each mouse. While the liver in situ tumor model (HCC) of comparative example 1 was a left abdominal surgical incision in mice, 25. mu.L of 1X 10 suspended in PBS6Individual Hepa1-6 cells were injected into the exposed liver of each mouse.
After the tumor reached a certain size, 5mg/kg of a rabbit Anti-mouse monoclonal VEGF-specific neutralizing antibody (cat # BD0801, supplied by pioneer pharmaceutical company (south kyo, jiangsu, china, hereinafter abbreviated as "Anti-VEGF neutralizing antibody" or "Anti-VEGF") as an Anti-vascular drug used in the present application was intraperitoneally injected into a part of tumor mice 2 times a week, and the remaining tumor mice were injected with a non-immune rabbit IgG isotype (cat # 10500C, Invitrogen, hereinafter abbreviated as "non-immune antibody NIIgG") as a control. Three weeks later, mice were dosed with a lethal dose of CO2Sacrifice, dissect and perform histological examination, RNA and protein extraction, hypoxia, apoptosis, etc.
FIG. 1 is a graph showing the examination of the resistance of various solid tumors to the anti-vascular drugs in comparative example 1. Wherein, CRC represents colon cancer, PDAC represents pancreatic cancer, HCC represents liver cancer, and WAT represents white fat area.
In the first row, panels a represent tumor volume studies after CRC tumor vaccination in the non-fat environment of healthy mice, treated with Anti-VEGF neutralizing antibody (Anti-VEGF) or non-immune antibody NIIgG, and panels B represent tumor volume changes after CRC tumor vaccination in the fat environment of WAT, treated with Anti-VEGF neutralizing antibody or non-immune antibody NIIgG; panel C represents the change in tumor volume following PDAC tumor inoculation in the non-fat environment of healthy mice, treated with anti-VEGF neutralizing antibody or non-immune antibody NIIgG, and panel D represents the change in tumor volume following PDAC tumor inoculation in the fat environment, treated with anti-VEGF neutralizing antibody or non-immune antibody NIIgG.
In the second row, the E-plot is the histological examination of healthy liver mice or fatty liver mice after construction of a CRC tumor liver metastasis model, after treatment with an anti-VEGF neutralizing antibody or non-immune NIIgG. The F picture shows the histological examination of non-fatty liver mice or fatty liver mice after the HCC liver tumor model is constructed and after the treatment of anti-VEGF neutralizing antibody or non-immune NIIgG.
Taking the A, B comparison of CRC tumors in FIG. 1 as an example, in the non-fat environment, the Anti-vascular drug Anti-VEGF can better inhibit the volume of CRC tumors, while in the fat environment, the Anti-VEGF and non-immune NIIgG have no significant difference in the change of tumor volume, which indicates that the CRC tumors are resistant to the Anti-vascular drug in the fat environment. Similarly, in panel E of figure 1, resistance to anti-vascular drugs has also occurred in CRC tumor mice with fatty liver. This suggests that the fat environment promotes the development of resistance to anti-angiogenic drugs by many common tumors such as CRC, PDAC (pancreatic cancer), HCC (liver cancer), etc.
Example 1: anti-vascular drug and CPT1a shRNA transfection vector for co-treating tumor in fat environment
Firstly, constructing a CPT1a shRNA transfection vector: cpt1shRNA lentiviral particles transfected with Cpt1shRNA were used as CPT1a shRNA transfection vectors, purchased from Santa Cruz Biotechnology (California). The transduction process was performed according to the manufacturer's protocol provided by the company, and CPT1a shRNA transfection vector could inhibit the expression of CPT1 protein.
For at about 1 × 105Density of cells/well MC38 and Hepa1-6 cells grown in 12-well plates were used 5X 10 cells, respectively4Transfection of Cpt1shRNA with Cpt1shRNA Lentiviral particles. Cells transduced with lentiviruses were then selected using puromycin (2 μ g/mL) and the knockdown efficiency of CPT1 was verified by standard quantitative real-time polymerase chain reaction (qPCR) methods.
In constructing a specific model of CRC liver metastases, MC38 cells transfected with the above lentiviruses or empty vector (as a control) were implanted from the liver of fatty liver mice, left abdominal surgery incisions were made, and 25. mu.L of 1X 10 cells suspended in PBS were placed6Each MC38 cell was injected into the exposed liver of each mouse and fed for a period of time to allow the tumor to reach a certain size, completing the construction of CRC liver metastases model.
In constructing a specific HCC liver in situ tumor model, the vector transfected or empty by the above lentiviruses was transfected (as a control)) Hepa1-6 cells were implanted from the liver of fatty liver mice, left abdominal surgery incisions were made, and 25. mu.L of 1X 10 cells suspended in PBS were placed6Each mouse was injected with Hepa1-6 cells and fed for a period of time to achieve a tumor size, completing the construction of a liver in situ tumor model (HCC).
The Anti-VEGF neutralizing antibody (Anti-VEGF) at 5mg/kg was intraperitoneally injected 2 times a week into tumor mice while using the non-immune antibody NIIgG as a control; three weeks later, mice were dosed with a lethal dose of CO2Sacrifice, dissect and perform histological examination, RNA and protein extraction, hypoxia, apoptosis, etc.
FIG. 2 is a graph showing the improvement effect of CPT1a on the resistance to an anti-vascular drug in a colon cancer (CRC) tumor and a liver in situ tumor (HCC) model in a fatty liver mouse in example 1 of the present invention.
As seen from fig. 2, in the adipose environment, for CRC tumors (a in fig. 2), the resistance of the anti-vascular drug was improved after using the CPT1a shRNA vector. In HCC tumors (B in fig. 2), a similar phenomenon was also observed. This indicates that the CPT1 inhibitor based on CPT1a shRNA inhibits the expression of CPT1a in tumor cells, can effectively restore the sensitivity of Anti-vascular drug Anti-VEGF, and reduce the tumor volume.
Example 2: co-treatment of tumors with anti-angiogenic drugs in adipose environment and CPT1a inhibitor drug carriers
Construction of CPT1a inhibitor drug carriers: the method comprises the following specific steps: etomoxir (Etomoxir) was dissolved in water to give a CPT1a inhibitor drug carrier solution.
In constructing the cell model, MC38 cells were seeded in 96-well plates at a density of 2500 cells/well and incubated in 100 μ L of the above medium for 24 hours, and each well was provided with an anaerobic environment using an anaerobic kit (tumor cells would use free fatty acids in an anaerobic environment), and divided into the following groups on day 3: neither Free Fatty Acid (FFA) nor Etomoxir was added (negative control); adding only FFA; FFA and 50. mu.M Etomovir were added; FFA and 100. mu.M Etomovir were added. Tumor cell growth was assessed in each well using the MTT assay system described above. Metabolic measurements of tumor cells were determined using a hippocampal metabolic analyzer and basal Oxygen Consumption Rate (OCR) using XF24 analyzer. The test results are shown in fig. 3.
In constructing a colon cancer (CRC) liver metastatic tumor model, MC38 cells were implanted from the liver of a fatty liver mouse, a left abdominal surgical incision was made, and 25. mu.L of 1X 10 cells suspended in PBS were placed6The MC38 cells are injected into the exposed liver of each mouse and fed for a period of time to make the tumor reach a certain size, thus completing the construction of liver metastasis tumor model. The 5mg/kg Anti-VEGF neutralizing antibody (Anti-VEGF) was intraperitoneally injected 2 times a week into tumor mice while using the non-immune antibody NIIgG as a control and 15mg/kg of the Etomovir aqueous solution was intraperitoneally injected every 15 days for two consecutive weeks (100. mu.L of Clodronate Liposomes (dichloromethylene bisphosphonate; Cloronate Liposomes, Netherlands, macrophage clearing reagent) were intravenously injected every 4 days during the experimental period for macrophage elimination) and sterile water was used as a negative control vehicle; after 2 weeks, mice were dosed with a lethal dose of CO2Sacrifice, dissect and perform histological examination, RNA and protein extraction, hypoxia, apoptosis, etc.
As shown in fig. 3 (a), the addition of Free Fatty Acid (FFA) significantly promoted the growth of CRC tumor cells in an adipose environment in the absence of oxygen. And when the CPT1a inhibitor drug Etomovir is used, the growth of tumor cells can be obviously inhibited. As shown in fig. 3 (b), the tumor cells showed an increase in metabolic rate after FFA addition (on the OCR curve, FCCP was added to the AntA stage), whereas the use of CPT1a inhibitor drug could inhibit tumor metabolism. This shows that under the anoxic environment, the additional addition of Free Fatty Acid (FFA) to the tumor cells can significantly promote the growth and metabolic activity of the tumor cells, and the inhibition of CPT1a can inhibit the growth and metabolic activity of the tumor cells.
FIG. 4 is a graph showing the results of examining the improvement of the resistance of the CPT1a inhibitor against the anti-vascular drug in the colon cancer (CRC) liver metastatic tumor model of the fatty liver mouse in example 2.
As seen in FIG. 4, in the CRC tumor liver metastasis model, the drug Etomovir, the CPT1a inhibitor, can effectively reduce the drug resistance of the tumor to the Anti-vascular drug Anti-VEGF under the condition of using the Anti-vascular drug Anti-VEGF; this demonstrates that using an etomoxider-based inhibitor of CPT1, it is effective in restoring the sensitivity of anti-vascular drugs and reducing tumor volume.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (7)
- Use of an inhibitor of CPT1a in the manufacture of a medicament for the treatment of a solid tumour that is resistant to an antiangiogenic drug; wherein the anti-vascular drug comprises a VEGF anti-angiogenic drug; the solid tumor with drug resistance to the antiangiogenic drug comprises one or more of liver cancer, lung cancer, pancreatic cancer, kidney cancer, stomach cancer, esophagus cancer, colon cancer, bladder cancer, breast cancer, ovarian cancer and cervical cancer; the CPT1a inhibitor acts on CPT1a on the surface of the mitochondrial outer membrane of the tumor cell with drug resistance to the antiangiogenic drugs, inhibits fatty acid from entering the mitochondrial matrix to be oxidized, inhibits the uptake of free fatty acid by the tumor cell, restores the sensitivity of the tumor cell to the antiangiogenic drug therapy, inhibits the proliferation of the tumor cell and promotes the apoptosis of the tumor cell.
- 2. The use of claim 1, wherein the CPT1a inhibitor comprises one or more of a chemical drug, a polypeptide drug, a protein drug and a gene drug that inhibits CPT1 a.
- 3. The use of claim 2, wherein said chemical that inhibits an inhibitor of CPT1a comprises etomoxir.
- 4. The use of claim 1, wherein the medicament for treating an anti-angiogenic drug resistant solid tumor further comprises a pharmaceutically acceptable carrier; the pharmaceutically acceptable carrier comprises at least one of a solvent, a polymer, a liposome, a recombinant viral vector and a eukaryotic recombinant expression vector.
- 5. The use as claimed in claim 1, wherein the CPT1a inhibitor comprises as a single active ingredient or with other pharmaceutically acceptable active ingredients an active ingredient in the medicament for the treatment of solid tumors that are resistant to antiangiogenic drugs.
- 6. A medicament for treating a solid tumor, the medicament comprising a CPT1a inhibitor and an anti-vascular drug; the anti-vascular drugs include VEGF anti-angiogenic drugs.
- 7. The medicament for treating solid tumors according to claim 6, wherein the mass ratio of the CPT1a inhibitor to the antiangiogenic medicament is (1-100): 1.
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