Bionic drug system with effect of cooperatively regulating lactic acid metabolism and application of bionic drug system in combination with PD-L1
By using a nanocomposite of MCT1 selective inhibitor and lactate oxidase in a biomimetic drug system to synergistically regulate lactate metabolism, the problem of poor therapeutic effect of PD-1/PD-L1 inhibitors is solved, the therapeutic effect on triple-negative breast cancer is enhanced, and the immune response is activated to inhibit tumor recurrence.
Patent Information
- Application Number
- CN202511858582.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-10
- Publication Date
- 2026-05-08
AI Technical Summary
Existing PD-1/PD-L1 inhibitors have limited efficacy in treating triple-negative breast cancer, mainly due to the acidic environment caused by lactic acid buildup in the tumor microenvironment and immunosuppression. Using MCT1 inhibitors such as AZD3965 alone can cause side effects and affect sensitivity.
The biomimetic drug system, including a selective MCT1 inhibitor and lactate oxidase, is used to form a nanocomplex via a biomimetic liposome carrier. This nanocomplex is precisely delivered to the tumor, preventing lactate efflux and promoting its consumption. Combined with a PD-L1 inhibitor, the therapeutic effect is enhanced.
It significantly improved the therapeutic effect of PD-L1 inhibitors, enhanced the tumor inhibition rate, and showed the potential to inhibit tumor recurrence in animal experiments, while improving the tumor's acidic immunosuppressive microenvironment.
Smart Images
Figure CN121987799A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, specifically relating to a biomimetic drug system with synergistic regulatory effect on lactic acid metabolism and its application in combination with PD-L1. Background Technology
[0002] Breast cancer ranks first among all cancers in women in my country. Among them, triple-negative breast cancer (TNBC) is characterized by its late clinical stage, high malignancy, strong invasiveness, early metastasis, easy recurrence and poor prognosis, making it the focus and challenge of breast cancer treatment. Recent studies have shown that PD-1 / PD-L1 antibodies need to be combined with basic chemotherapy to be effective in treating advanced triple-negative breast cancer. However, the efficacy rate of PD-1 / PD-L1 inhibitors in tumor treatment is less than 50%, which is largely related to the immunosuppressive effect of the tumor microenvironment. Even under aerobic conditions, tumor cells prefer glycolysis for energy. Lactic acid is the main metabolite of glycolysis, and its excessive accumulation in the tumor microenvironment leads to a weakly acidic environment in most solid tumor tissues, inducing an acidic tumor microenvironment and causing immunosuppression. TNBC is a solid tumor that relies on glycolysis for energy, resulting in lactic acid accumulation in the extratumoral fluid, thus limiting the therapeutic effect of PD-1 / PD-L1 inhibitors.
[0003] Monocarboxylic acid transporter 1 (MCT1) is a member of the monocarboxylic acid transporter family and plays an important role in tumor pH regulation and energy metabolism. AZD3965 is a selective inhibitor of MCT1. Studies have shown that AZD3965 can effectively inhibit lactate transport in tumors, increase intracellular lactate concentration, and thus inhibit tumor cell growth. However, clinical observations have revealed that intracellular free AZD3965 can cause various side effects. Literature review, "In Vitro and In Vivo Efficacy of AZD3965 and Alpha-Cyano-4-Hydroxycinnamic Acid in the Murine 4T1 Breast Tumor Model," found that AZD3965 alone can reduce the expression of the tumor proliferation marker Ki67, but increases intracellular lactate concentration in tumor cells. Further research has shown that the accumulation of intracellular lactate may induce the upregulation of MCT4 expression, and the upregulation of MCT4 expression may, in turn, affect the sensitivity of MCT1 inhibition.
[0004] Therefore, new methods and strategies are needed to alleviate the shortcomings of existing technologies. Summary of the Invention
[0005] The purpose of this invention is to provide a biomimetic drug system with synergistic regulatory effect on lactate metabolism and its application in combination with PD-L1 inhibitors in the field of tumor treatment, thereby partially solving or alleviating the above-mentioned deficiencies in the prior art. The invention specifically adopts the following technical solutions.
[0006] A first aspect of the present invention is to provide a novel pharmaceutical system having the function of regulating lactic acid metabolism.
[0007] A biomimetic drug system with synergistic regulatory effect on lactate metabolism, the biomimetic drug system comprising a drug combination for regulating lactate metabolism and a carrier; the drug combination consists of an MCT1 selective inhibitor and lactate oxidase; the carrier is a biomimetic liposome incorporating erythrocyte membrane proteins.
[0008] Furthermore, the drug combination that regulates lactate metabolism and the carrier form a nano-like complex.
[0009] Furthermore, the selective inhibitors of MCT1 include AZD3965, AR-C155858, BAY-8002, or 7ACC2.
[0010] Furthermore, the mass ratio of the MCT1 selective inhibitor to lactate oxidase in the drug combination ranges from 1:1 to 5:1.
[0011] As a preferred embodiment, the mass ratio of the MCT1 selective inhibitor to lactate oxidase in the drug combination is 5:1.
[0012] Furthermore, the biomimetic liposome contains 0.5% to 2% erythrocyte membrane proteins by mass.
[0013] As a preferred embodiment, the biomimetic liposome contains 2% red blood cell membrane proteins by mass.
[0014] Another aspect of the present invention is to provide a novel drug combination for treating tumors.
[0015] A combination therapy for treating solid tumors, the combination therapy comprising the aforementioned biomimetic drug system with synergistic regulatory effects on lactate metabolism and a PD-L1 inhibitor.
[0016] As a preferred embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody.
[0017] Furthermore, the mass ratio of the biomimetic drug system with synergistic regulatory effect on lactic acid metabolism to the PD-L1 inhibitor ranges from 1:1 to 2:1.
[0018] As a preferred embodiment, the mass ratio of the biomimetic drug system with synergistic regulatory effect on lactate metabolism to the PD-L1 inhibitor is 2:1.
[0019] In a specific embodiment of the present invention, AZD3965 is 2 mg / kg and PD-L1 inhibitor is 1 mg / kg, but this is not a limitation.
[0020] Furthermore, the solid tumors mentioned include breast cancer, but are not a limitation.
[0021] Another aspect of the present invention is to provide the application of the above-described pharmaceutical system that regulates lactic acid metabolism.
[0022] The application of a biomimetic drug system with synergistic regulatory effect on lactate metabolism in the preparation of a potency enhancer for PD-L1 inhibitors, the biomimetic drug system comprising a drug combination for regulating lactate metabolism and a carrier; the drug combination consists of an MCT1 selective inhibitor and lactate oxidase; the carrier is a biomimetic liposome incorporating erythrocyte membrane proteins.
[0023] Furthermore, the biomimetic drug system with synergistic regulatory effect on lactate metabolism is used to enhance the inhibitory effect of the PD-L1 inhibitor on solid tumors.
[0024] Furthermore, the potency enhancer of the PD-L1 inhibitor is an experimental reagent, mainly used for scientific research and preclinical drug testing.
[0025] Beneficial technical effects: This invention first provides a drug system with lactate metabolism-regulating effects. The system comprises a combination of drugs regulating lactate metabolism and a carrier, with the carrier encapsulating the drugs to form a stable nano-like complex. This drug system can precisely deliver MCT1 inhibitors and LOX into solid tumors, improving the tumor's slightly acidic immunosuppressive microenvironment by inhibiting intracellular lactate efflux and promoting the consumption of accumulated lactate. Furthermore, this invention also provides a combination of the aforementioned lactate metabolism-regulating drug system with PD-L1 inhibitors. Experiments have demonstrated that this combination synergistically enhances the therapeutic effect of PD-L1 inhibitors. In animal experiments, the combination group significantly increased tumor inhibition rates, and this combination also shows potential in inhibiting tumor recurrence. Attached Figure Description
[0026] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.
[0027] Figure 1 The prepared drug-loaded biomimetic liposomes are characterized in one embodiment of the present invention; Figure 2 This invention demonstrates the cellular uptake and in vitro antitumor efficacy of drug-loaded biomimetic liposomes in one embodiment. Figure 3 This is an example of the in vitro lactate metabolism regulation and ICD induction effect of drug-loaded biomimetic liposomes in one embodiment of the present invention; Figure 4 This is the result of treating a mouse model of breast cancer with drug-loaded biomimetic liposomes combined with PD-L1 antibody in one embodiment of the present invention; Figure 5 This is a schematic diagram of the biomimetic drug system with synergistic regulatory effect on lactic acid metabolism prepared according to the present invention. Detailed Implementation
[0028] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.
[0029] In this document, "and / or" includes any and all combinations of one or more of the listed related items.
[0030] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.
[0031] As used in this specification, the term "about" typically means + / -5% of the value, more typically + / -4% of the value, more typically + / -3% of the value, more typically + / -2% of the value, even more typically + / -1% of the value, and even more typically + / -0.5% of the value.
[0032] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.
[0033] Detailed description of the attached images: Figure 1 (A) Particle size and transmission electron microscopy images of Lipo, Blipo, and AZD / LOX@Blipo. Scale bar: 50 nm. (B) Zeta potential of Lipo, Blipo, and AZD / LOX@Blipo. (C) Changes in hydrated particle size, PDI, and zeta potential over 5 days. (D) SDS-PAGE analysis of LOX, membrane proteins, Lipo, Blipo, and AZD / LOX@Blipo. (E) Wheat germ lectin assay showing the presence of glycosylated proteins on the surface of biomimetic liposomes. (F) Cumulative release behavior of AZD3965 and AZD / LOX@Blipo in PBS at different pH values. Data are expressed as mean ± SD (n=3). **p<0.01, ***p<0.001.
[0034] Figure 2 (A) Cellular uptake of biomimetic liposomes with different membrane protein contents (Blipo1, Blipo2, and Blipo3 represent protein contents of 0.5%, 1%, and 2%, respectively). (B) Cellular uptake of Blipo3 at 2 h and 4 h. (C) In vitro cytotoxicity of 4T1 cells after co-incubation with LOX, AZD, LOX@Blipo, AZD@Blipo, and AZD / LOX@Blipo for 24 h and 48 h. (DE) Cell apoptosis / necrosis after 6 h of treatment in different experimental groups. (F) Live / dead cell staining images of 4T1 cells after 6 h of treatment in different experimental groups. Scale bar: 100 μm. Data are expressed as mean ± standard deviation (n=3). *p<0.05, **p<0.01, ***p<0.001.
[0035] Figure 3(A) Lactate content in the culture medium after co-culturing 4T1 mouse triple-negative breast cancer cells with different experimental groups of drugs. (B) Lactate inhibition rate after treatment in different experimental groups. (C) H2O2 content and (D) GSH level. (E) Intracellular pH value detected by BCECF-AM staining; scale bar: 10 μm. Fluorescence images (F) and flow cytometry analysis (G) of intracellular ROS levels after treatment in different experimental groups; scale bar: 75 μm. CLSM images (H) and ELISA analysis (I) of HMGB1 release. CLSM images (J) and flow cytometry analysis (K) exposed by CRT; scale bar: 10 μm. (L) ATP release after different treatments. Data are expressed as mean ± standard deviation (n=3). *p<0.05, **p<0.01, ***p<0.001.
[0036] Figure 4 (A) Schematic diagram of the 4T1 tumor-bearing mouse model and combined treatment regimen. (B) Average body weight of mice after treatment with different drugs. (C) Tumor volume curves after treatment with different drugs. (D) Tumor weight after treatment with different drugs. (E) Photographs of tumors after treatment with different drugs; scale bar: 1 cm. Flow cytometry was used to analyze CD3+ in tumor tissue. + CD8 + T cells (F), IFN-γ + CD8 + T cells (G), TNF-α + CD8 + T cells (H), Treg cells CD4 + CD25 + FoxP3 + (I) proportion; CD44 effector memory T cells in the spleen + CD62L - CD4 + (J) and CD44 + CD62L - CD8 + The proportion (K). Data are expressed as mean ± standard deviation (n=5). *p<0.05, **p<0.01, ***p<0.001.
[0037] Definition of noun: The "bionic" in this invention refers to embedding and compounding red blood cell membrane proteins into ordinary liposomes to give them higher affinity for tumor cells.
[0038] The “synergy” described in this invention includes synergy within the biomimetic drug system and synergy between the biomimetic drug system and the PD-L1 inhibitor.
[0039] The "synergy within the biomimetic drug system" refers to the fact that the drug combination (AZD3965+LOX) it contains can simultaneously prevent lactic acid efflux and consume lactic acid accumulation within tumor cells, thereby synergistically achieving a significant tumor-inhibiting effect.
[0040] The "synergy between the biomimetic drug system and the PD-L1 inhibitor" refers to the biomimetic drug system enhancing the efficacy of the PD-L1 inhibitor by regulating lactate within the tumor and activating immunity.
[0041] Example 1 Constructing a drug-loaded biomimetic liposome carrying a monocarboxylic acid transporter 1 (MCT1) inhibitor and lactate oxidase (LOX) (AZD / LOX@Blipo) The preferred MCT1 selective inhibitor in this embodiment is AZD3965. However, those skilled in the art will understand that other MCT1 selective inhibitors, such as AR-C155858, BAY-8002, and 7ACC2, are also within the scope of this invention.
[0042] First, erythrocyte membranes were extracted using a hypotonic hemolysis method. The extracted erythrocyte membranes were then treated with a cell membrane protein extraction reagent and stored at -80 ℃ to obtain erythrocyte membrane proteins (hereinafter referred to as membrane proteins).
[0043] Liposomes were then prepared by thin-film hydration.
[0044] Then, the erythrocyte membrane proteins and liposomes are extruded through a liposome extruder to obtain a biomimetic carrier, or biomimetic liposome (membrane protein + liposome).
[0045] The drug was then loaded onto a biomimetic carrier; in this example, the drug was a combination of AZD3965 and LOX, resulting in the biomimetic drug system AZD / LOX@Blipo (hereinafter referred to as the biomimetic drug system), which has a synergistic regulatory effect on lactate metabolism. The prepared biomimetic drug system was characterized, and the results are shown below. Figure 1 .
[0046] The results showed that the average particle size of the prepared biomimetic liposome Blipo was 84.57 ± 0.88 nm, while the average particle size of the ordinary liposome Lipo was 94.81 ± 0.75 nm, and the average particle size of the biomimetic drug system AZD / LOX@Blipo was 81.98 ± 0.99 nm. Loading AZD3975 and LOX had no significant effect on the size and morphology of the biomimetic liposomes; all three types of liposomes were spherical. Figure 1A). All three types of liposomes exhibited negative surface potentials. Compared to the ordinary liposome group (-17.6 mV), the potential of the biomimetic liposomes decreased to approximately -22 mV, suggesting that the integration of erythrocyte membrane proteins into the liposomes endowed the ordinary liposomes with biomimetic functions. Figure 1 B). After incubating the three liposomes in water for 5 days, no significant changes were observed in their particle size, PDI, or surface potential. Figure 1 C). The fusion of Blipo and AZD / LOX@Blipo proteins was detected by SDS-PAGE, and the results are as follows: Figure 1 As shown in Figure D, the bands in lanes 5 and 6 (Blipo and AZD / LOX@Blipo) are partially identical to the band in lane 3 (erythrocyte membrane proteins), indicating that the biomimetic liposomes contain erythrocyte membrane proteins. The WGA fluorescence intensity of the biomimetic liposome group is 2.3 times that of the liposomes, verifying the presence of glycosylated proteins on the surface of the biomimetic liposomes, indicating that the membrane proteins are correctly oriented during integration. Figure 1 E). Finally, the drug release of AZD3965 is as follows: Figure 1 As shown in Figure F. Under acidic or neutral conditions, the biomimetic drug system exhibits a slow drug release pattern over 72 hours. The structural relationships of the biomimetic drug system are shown in Figure F. Figure 5 Among them, AZD3965 is distributed in the middle of the phospholipid bilayer of the liposome; LOX is located in the central aqueous phase of the liposome; and erythrocyte membrane proteins are embedded in the phospholipid bilayer.
[0047] Example 2 Investigating the in vitro antitumor activity and lactate-regulating ability of AZD / LOX@Blipo Biomimetic liposomes with different membrane protein contents (Blipo1, Blipo2, and Blipo3 represent membrane protein / lipid ratios of 0.5%, 1%, and 2%, respectively) were prepared. The changes in the uptake of biomimetic liposomes by tumor cells were investigated by flow cytometry to screen for the optimal membrane protein integration ratio. Subsequently, the cytotoxicity of the biomimetic drug system prepared according to the optimal membrane protein ratio was verified.
[0048] Furthermore, this biomimetic drug system includes an MCT1 inhibitor and lactate oxidase. The MCT1 inhibitor inhibits lactate efflux, while LOX catalyzes lactate production and generates hydrogen peroxide and reactive oxygen species (ROS). Subsequent investigations examined changes in tumor cell pH, lactate, hydrogen peroxide, glutathione, and ROS levels. ICD (intracytoplasmic reticulum death) was induced by ROS, and ATP and HMGB1 release, as well as CRT protein exposure, were then investigated. Results are shown in [Figure number missing]. Figure 2 .
[0049] The results showed that, Figure 2As shown in Figure A, the uptake of liposomes by 4T1 cells increases with the increase of membrane protein content. This suggests that the incorporation of membrane proteins increases the affinity of ordinary liposomes for tumor cells, giving them biomimetic functions. In preliminary experiments, our research team found that when the mass ratio of erythrocyte membrane proteins in the biomimetic carrier increased to more than 2%, the uptake level of tumor cells did not increase significantly. Therefore, we determined that the mass ratio of membrane proteins in the biomimetic liposomes should be 2%.
[0050] Liposomes were labeled with another fluorescent dye, and cellular uptake of conventional liposomes and biomimetic liposomes (Blipo3) was observed at 2 h and 4 h. Figure 2 As shown in Figure B, after incubation for 2 h or 4 h, the DiR fluorescence intensity of Blipo was higher than that of ordinary liposomes. This confirms that the biomimetic liposomes integrating erythrocyte membrane proteins can improve the uptake efficiency of tumor cells to a certain extent. The above results demonstrate that the biomimetic liposomes prepared in this invention have certain tumor targeting properties.
[0051] Cell viability was assessed using the CCK8 assay after 24 h and 48 h of treatment with free drug groups (LOX, AZD), drug systems loaded with different single drugs, and drug systems loaded with two drugs (LOX@Blipo, AZD@Blipo, AZD / LOX@Blipo). Figure 2 As shown in Figure C, free AZD3965 had no significant effect on tumor cell survival, while LOX treatment showed a certain inhibitory effect. However, among the experimental groups, AZD / LOX@Blipo exhibited the strongest killing and inhibitory effects on tumor cells, demonstrating that this approach, which simultaneously blocks lactate efflux and consumes lactate intracellularly, effectively regulates lactate metabolism within tumor cells, thereby enhancing the tumor-killing effect.
[0052] To further investigate its antitumor activity, flow cytometry was used to analyze the apoptosis of tumor cells treated with different experimental groups. The results showed that the apoptosis rate was highest (38.93%) after 6 h of treatment with AZD / LOX@Blipo. Figure 2 D). Observation of fluorescence images of live / dead cells showed results consistent with CCK8 and apoptosis experiments, indicating that the biomimetic drug system loaded with two drugs induced the highest proportion of cell death. Figure 2 F). The above results demonstrate that the biomimetic drug system prepared in this invention exhibits significant in vitro antitumor activity through synergistic regulation of lactic acid metabolism.
[0053] Example 3 To investigate the synergistic lactate-regulating effect and the effect of AZD / LOX@Blipo in inducing immunogenic cell death (ICD).
[0054] First, the inhibitory effects of different experimental groups (LOX group, AZD group, LOX@Blipo group, AZD@Blipo group, AZD / LOX@Blipo) on lactate efflux were investigated. Figure 3 As shown in Figures AB, the control group had the highest lactate content; the lactate inhibition rate (reduction in lactate secretion) after treatment with AZD3965 alone was 39.5%; the lactate inhibition rate after treatment with AZD@Blipo was 52.8%, and the lactate inhibition rate after treatment with LOX@Blipo was 51.7%; the lactate inhibition rate after treatment with AZD / LOX@Blipo was 65.0%, the highest among all experimental groups.
[0055] When lactate levels decrease, the pH value inside tumor cells increases. For example... Figure 3 As shown in Figure E, the AZD / LOX@Blipo group exhibited the strongest green fluorescence. LOX produced significant amounts of H2O2 and pyruvate while consuming lactic acid. Although both LOX and LOX@Blipo treatments increased H2O2 production, the highest H2O2 production was observed in the AZD / LOX@Blipo group. Figure 3 (C) This demonstrates that the combined use of AZD3965 and LOX produced a synergistic effect in regulating lactate levels.
[0056] Figure 3 D shows the results of downregulation of intracellular GSH levels in each experimental group. ROS levels are as follows: Figure 3 As shown in FG, the flow cytometry quantitative results are consistent with the fluorescence image results, and the AZD / LOX@Blipo group has the strongest ability to induce ROS production.
[0057] Further investigation was conducted on ICD-related indicators including calreticulin (CRT), high-mobility group box 1 (HMGB1), and adenosine triphosphate (ATP). Figure 3 As shown in HI, AZD / LOX@Blipo can induce more HMGB1 release from the nucleus into the extracellular matrix. Both CRT fluorescence imaging and flow cytometry results indicate that AZD / LOX@Blipo treatment induces greater CRT exposure (…). Figure 3 JK). Meanwhile, damaged cells treated with AZD / LOX@Blipo released a large amount of ATP ( Figure 3 In summary, AZD / LOX@Blipo-mediated synergistic lactate regulation and ROS trigger stronger oxidative stress, leading to enhanced ICD effects and ultimately increasing tumor immunogenicity.
[0058] Example 4 Cancer treatment efficacy of AZD / LOX@Blipo in combination with PD-L1 inhibitor The animal model used in this invention is the 4T1 mouse orthotopic breast cancer model. Compared with other subtypes of breast cancer, TNBC exhibits broader PD-L1 expression. In this embodiment, AZD / LOX@Blipo combined with PD-L1 antibody (Anti-mouse PD-L1 (B7-H1)-InVivo, Selleck) was selected.
[0059] The therapeutic effect of drug-loaded biomimetic liposome AZD / LOX@Blipo combined with a PD-L1 inhibitor (hereinafter referred to as αPD-L1) on tumors was investigated using a 4T1 orthotopic breast cancer mouse model. Once the tumor volume reached a certain size, tumor-bearing mice were randomly divided into four groups of five mice each: a saline group, an αPD-L1 group, an AZD / LOX@Blipo group, and an AZD / LOX@Blipo+αPD-L1 group. AZD / LOX@Blipo was administered once every four days for three consecutive times, and αPD-L1 was administered one day after the drug-loaded biomimetic liposome (three times in total). Tumor growth and changes in body weight were observed. After treatment, changes in the immune microenvironment in tumor tissue and the proportion of memory T cells in the spleen were examined by flow cytometry. The experimental procedure is described below. Figure 4 A.
[0060] The results showed that, compared with the saline group, αPD-L1 monotherapy resulted in rapid tumor growth and limited tumor suppression effect. Figure 4 (CE). Tumor volume was significantly reduced in the AZD / LOX@Blipo group, but the tumor inhibition rate was highest in the AZD / LOX@Blipo combined with αPD-L1 group (68.2%), which was higher than the 49.8% in the group with a single dual-drug biomimetic drug system. These results demonstrate that AZD / LOX@Blipo and PD-L1 antibody produced a synergistic anti-tumor effect. Flow cytometry was used to detect the expression of relevant lymphocytes in the spleen and tumor tissue to further evaluate the treatment efficacy. Figure 4 FH analysis showed that the percentage of tumor-infiltrating lymphocytes producing IFN-γ and TNF-α was increased in the combination therapy group, indicating that the T-cell response validated the antitumor effect of the combination therapy. Furthermore, compared with the control group, the percentage of Treg cells was significantly reduced ( Figure 4 I). Notably, the combination therapy group also activated spleen effector memory T (TEM) cells (CD4+). + CD62L - CD44 + and CD8 + CD62L - CD44 + () Figure 4 JK). Compared with mice treated with AZD / LOX@Blipo alone or αPD-L1 alone, the combination treatment group reduced CD8+. +The TEM cell population increased by 1.5-fold and 2.4-fold, respectively. The combination of AZD / LOX@Blipo and αPD-L1 shows potential in inhibiting tumor recurrence. This is because the level of TEM cells in the spleen reflects the strength and readiness of the systemic anti-tumor immune response. Elevated TEM cell counts are direct evidence of the body's successful and effective immune response to tumor antigens, forming long-term immune memory. Strong immune memory is central to preventing tumor recurrence.
[0061] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.
Claims
1. A biomimetic drug system with synergistic regulatory effect on lactic acid metabolism, characterized in that, The biomimetic drug system includes a drug combination for regulating lactate metabolism and a carrier; the drug combination consists of an MCT1 selective inhibitor and lactate oxidase; the carrier is a biomimetic liposome incorporating erythrocyte membrane proteins.
2. The biomimetic drug system as described in claim 1, characterized in that, The selective inhibitors of MCT1 include AZD3965, AR-C155858, BAY-8002, or 7ACC2.
3. The biomimetic drug system as described in claim 1, characterized in that, The mass ratio of the MCT1 selective inhibitor to lactate oxidase in the drug combination is 1:1 to 5:
1.
4. The biomimetic drug system as described in claim 1, characterized in that, The biomimetic liposomes contain 0.5% to 2% erythrocyte membrane proteins by mass.
5. A combination drug regimen for treating solid tumors, characterized in that, The combination of drugs includes the biomimetic drug system with synergistic regulatory effect on lactate metabolism as described in any one of claims 1-4 and a PD-L1 inhibitor.
6. The combination drug regimen as described in claim 5, characterized in that, The PD-L1 inhibitors include anti-PD-L1 antibodies.
7. The combination drug regimen as described in claim 5, characterized in that, The mass ratio of the biomimetic drug system with synergistic regulatory effect on lactic acid metabolism to the PD-L1 inhibitor is 1:1 to 2:
1.
8. The combination of drugs as described in claim 5, characterized in that, The solid tumors include breast cancer.
9. The application of a biomimetic drug system with synergistic regulatory effect on lactate metabolism in the preparation of a potency enhancer for PD-L1 inhibitors, characterized in that, The biomimetic drug system includes a drug combination for regulating lactate metabolism and a carrier; the drug combination consists of an MCT1 selective inhibitor and lactate oxidase; the carrier is a biomimetic liposome incorporating erythrocyte membrane proteins.
10. The application as described in claim 9, characterized in that, The biomimetic drug system with synergistic regulatory effect on lactate metabolism is used to enhance the inhibitory effect of the PD-L1 inhibitor on solid tumors.