[go: up one dir, main page]

WO2025140008A1 - Utilisation d'une protéine ccl11 recombinante dans le traitement d'effusions pleurales malignes - Google Patents

Utilisation d'une protéine ccl11 recombinante dans le traitement d'effusions pleurales malignes Download PDF

Info

Publication number
WO2025140008A1
WO2025140008A1 PCT/CN2024/140718 CN2024140718W WO2025140008A1 WO 2025140008 A1 WO2025140008 A1 WO 2025140008A1 CN 2024140718 W CN2024140718 W CN 2024140718W WO 2025140008 A1 WO2025140008 A1 WO 2025140008A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
mpe
ccl11
eosinophils
subject
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/CN2024/140718
Other languages
English (en)
Chinese (zh)
Inventor
沈华浩
应颂敏
张敏
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang University ZJU
Original Assignee
Zhejiang University ZJU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhejiang University ZJU filed Critical Zhejiang University ZJU
Publication of WO2025140008A1 publication Critical patent/WO2025140008A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/195—Chemokines, e.g. RANTES
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/10—Antioedematous agents; Diuretics

Definitions

  • the present invention relates to the field of medicine, and in particular, to application of recombinant protein CCL11 in treating malignant pleural effusion.
  • MPE Malignant pleural effusions
  • MPE Malignant pleural effusions
  • MPE Malignant pleural effusions
  • MPE is a pleural effusion caused by malignant pleural mesothelioma originating in the pleural cavity or metastasis of tumors from other parts such as the lungs, gastrointestinal tract, and breast to the pleural cavity.
  • MPE is a poor prognostic sign for patients with malignant tumors.
  • the median survival of MPE patients is 3-12 months, with poor prognosis and high mortality rate, and clinical treatment is a huge challenge.
  • MPE patients mainly present with progressively worsening dyspnea, chest pain and dry cough, which seriously affect their quality of life.
  • the treatment for MPE mainly includes closed chest drainage and chemical/surgical pleurodesis, both of which are palliative treatments and cannot significantly improve the survival of patients.
  • the purpose of the present invention is to provide a medicine and its application for effectively treating and alleviating the complications of MPE.
  • a recombinant protein CCL11 or a nucleic acid molecule encoding the protein CCL11 for preparing a drug for treating and/or alleviating malignant pleural effusion (MPE) in a subject.
  • MPE malignant pleural effusion
  • the subject has one or more MPE typing characteristics selected from the following group:
  • the "significant increase” refers to the ratio of the amount or concentration C1 of CCL11 protein in the pleural effusion of the subject to the amount or concentration C0 of CCL11 protein in the pleural transudate (i.e., C1/C0), which is ⁇ 1.5, preferably ⁇ 2.0, and more preferably ⁇ 3.0.
  • the concentration of CCL11 protein in pleural effusion is ⁇ 800 pg/mL, preferably ⁇ 1000 pg/mL; more preferably ⁇ 1100 pg/mL.
  • the "significant decrease in the number of eosinophils” refers to the ratio of the number or concentration of eosinophils in the subject's pleural effusion, B1, to the number or concentration of eosinophils in the subject's own peripheral blood, B0 (i.e., B1/B0), which is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the "significant decrease in the number of eosinophils" refers to the ratio of the number or concentration D1 of eosinophils in the pleural effusion of the subject to the number or concentration D0 of eosinophils in the pleural transudate (i.e., D1/D0), which is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the "significantly upregulated expression” refers to the ratio of the expression level (mRNA or protein) M1 of the marker in the eosinophils of the subject to the expression level (mRNA or protein) M0 of the marker in the eosinophils in the subject's own bone marrow (i.e., M1/M0), which is ⁇ 1.5, preferably ⁇ 2.0, and more preferably ⁇ 3.0.
  • the "significantly downregulated expression” refers to the ratio of the expression level (mRNA or protein) N1 of the marker in the eosinophils of the subject to the expression level (mRNA or protein) N0 of the marker in the eosinophils in the subject's own bone marrow (i.e., N1/N0), ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the subject has one or more auxiliary typing features selected from the following group:
  • the subject's immune T cell function is intact.
  • the recombinant protein CCL11 recruits eosinophils into the chest cavity, thereby alleviating MPE through eosinophils.
  • the recombinant protein CCL11 is exogenous recombinant CCL11 (rCCL11).
  • the recombinant protein CCL11 is of human or mouse origin.
  • the MPE refers to pleural effusion caused by an in situ tumor or a metastatic tumor.
  • the in situ tumor is selected from the following group: lung cancer, breast cancer, or a combination thereof.
  • the metastatic tumor is selected from the group consisting of colon cancer, gastric cancer, large intestine cancer, pancreatic cancer, or a combination thereof.
  • the expression level of Ki67 on CD3+T cells in the pleural effusion of the subject is low; and/or the expression level of CD69 on CD3+T cells in the pleural effusion of the subject is low.
  • the "low expression level of Ki67” refers to the ratio of the expression level E1 of Ki67 on CD3+T cells in the pleural effusion of the subject to the expression level E0 of Ki67 on CD3+T cells in the subject's own peripheral blood (i.e., E1/E0), which is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the "low CD69 expression level” refers to the ratio of the expression level F1 of CD69 on CD3+T cells in the pleural effusion of the subject to the expression level F0 of CD69 on CD3+T cells in the subject's own peripheral blood (i.e., F1/F0), which is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the pleural effusion of the subject has low levels of NK cells, DC cells and mononuclear macrophages.
  • the “low content of NK cells, DC cells, and monocyte macrophages” refers to the ratio of the number of NK cells (numerator) in the pleural effusion of the subject to the number of NK cells (denominator) in the pleural transudate of the subject, Z1 ⁇ 0.75, preferably ⁇ 0.5, more preferably ⁇ 0.3;
  • the ratio Z2 of the number of DC cells (numerator) to the number of NK cells in the pleural transudate (denominator) is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3;
  • the ratio Z3 of the number of mononuclear macrophages (numerator) to the number of NK cells in the pleural effusion (denominator) is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the pleural effusion of the subject has low levels of ⁇ T cells, Th1 cells, and Th17 cells.
  • the “low content of ⁇ T cells, Th1 cells, and Th17 cells” refers to the ratio of the number of ⁇ T cells (numerator) in the pleural effusion of the subject to the number of ⁇ T cells (denominator) in the pleural transudate of the subject, Z4 ⁇ 0.75, preferably ⁇ 0.5, more preferably ⁇ 0.3;
  • the ratio Z5 of the number of Th1 cells (numerator) to the number of Th1 cells in the pleural transudate (denominator) is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3;
  • the ratio Z6 of the number of Th17 cells (numerator) to the number of Th17 cells in the pleural effusion (denominator) is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the subject's pleural effusion has a high content of Treg cells.
  • the "high Treg cell content” refers to that the ratio of the number of Treg cells (numerator) in the pleural effusion of the subject to the number of Treg cells (denominator) in the pleural exudate is ⁇ 1.2, preferably ⁇ 1.8, and more preferably ⁇ 2.5.
  • the treatment and/or relief refers to a reduction in the volume of pleural tumors and a reduction in the amount of pleural effusion.
  • the reduction in pleural effusion volume refers to the restoration of the subject's pleural effusion to a normal pleural effusion content (ie, no pleural effusion).
  • the pleural effusion of the subject has low levels of ⁇ T cells, Th1 cells and Th17 cells, and high levels of Treg cells.
  • the treatment and/or relief refers to that the content of ⁇ T cells, Th1 cells and/or Th17 cells in the subject's pleural lavage fluid is increased by ⁇ 50% compared with that before treatment; preferably, ⁇ 100%.
  • the treatment and/or relief refers to that the content of Treg cells in the chest lavage fluid of the subject is 80% of the content of Treg cells in the chest lavage fluid before treatment; preferably, 50%.
  • a pharmaceutical composition comprising (a) a recombinant protein CCL11 or a nucleic acid molecule encoding protein CCL11 as a first active ingredient; and (b) an immunostimulant as a second active ingredient; and (c) optionally a pharmaceutically acceptable carrier.
  • the immunostimulant is selected from the following group: interleukin-2, immune checkpoint inhibitors, CAR-T cells, or a combination thereof.
  • the immune checkpoint inhibitor is selected from the group consisting of CTLA4, PD-1, PD-L1, or a combination thereof.
  • the pharmaceutical composition is an injection.
  • the pharmaceutical composition is in the form of an intrathoracic administration.
  • the pharmaceutical composition according to the second aspect of the present invention in the preparation of a medicament for treating and/or alleviating MPE.
  • the drug is an injection.
  • the drug is in the form of an intrathoracic administration.
  • a method for preventing and/or treating and/or alleviating MPE comprising administering a therapeutically effective amount of recombinant protein CCL11 or a nucleic acid molecule encoding protein CCL11 to a subject in need thereof.
  • the method is non-diagnostic and non-therapeutic.
  • the subject is a mammal; preferably a mouse, a rat, a rabbit, a monkey or a human.
  • the expression level of Ki67 on CD3+T cells in the pleural effusion of the subject is low; and/or the expression level of CD69 on CD3+T cells in the pleural effusion of the subject is low.
  • the "low expression level of Ki67” refers to the ratio of the expression level E1 of Ki67 on CD3+T cells in the pleural effusion of the subject to the expression level E0 of Ki67 on CD3+T cells in the subject's own peripheral blood (i.e., E1/E0), which is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the "low CD69 expression level” refers to the ratio of the expression level F1 of CD69 on CD3+T cells in the pleural effusion of the subject to the expression level F0 of CD69 on CD3+T cells in the subject's own peripheral blood (i.e., F1/F0), which is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the eosinophil function of the subject is impaired or suppressed.
  • the pleural effusion of the subject has low levels of ⁇ T cells, Th1 cells, and Th17 cells.
  • the “low content of ⁇ T cells, Th1 cells, and Th17 cells” refers to the ratio of the number of ⁇ T cells (numerator) in the pleural effusion of the subject to the number of ⁇ T cells (denominator) in the pleural transudate of the subject, Z4 ⁇ 0.75, preferably ⁇ 0.5, more preferably ⁇ 0.3;
  • the ratio Z5 of the number of Th1 cells (numerator) to the number of Th1 cells in the pleural transudate (denominator) is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3;
  • the ratio Z6 of the number of Th17 cells (numerator) to the number of Th17 cells in the pleural effusion (denominator) is ⁇ 0.75, preferably ⁇ 0.5, and more preferably ⁇ 0.3.
  • the subject's pleural effusion has a high content of Treg cells.
  • the "high Treg cell content” refers to that the ratio of the number of Treg cells (numerator) in the pleural effusion of the subject to the number of Treg cells (denominator) in the pleural exudate is ⁇ 1.2, preferably ⁇ 1.8, and more preferably ⁇ 2.5.
  • the treatment and/or relief refers to a reduction in the volume of pleural tumors and a reduction in the amount of pleural effusion.
  • the pleural effusion of the subject has low levels of ⁇ T cells, Th1 cells, and Th17 cells, and high levels of Treg cells.
  • the treatment and/or relief refers to that the content of ⁇ T cells, Th1 cells and Th17 cells in the subject's pleural lavage fluid is increased by ⁇ 50% compared with that before treatment; preferably, ⁇ 100%.
  • the treatment and/or relief refers to that the content of Treg cells in the chest lavage fluid of the subject is 80% of the content of Treg cells in the chest lavage fluid before treatment; preferably, 50%.
  • Figure 1 shows the status of eosinophils in pleural lavage fluid after LLC modeling.
  • Panel A shows representative confocal microscopy images of eosinophils in pleural lavage fluid on day 3, stained with DAPI and EPX; scale bar: 20 ⁇ m.
  • Panel B shows the gating strategy of flow cytometry for mouse eosinophils. Eosinophils were labeled with DAPI-CD45+Gr1-/lowCD11c-CD11b+F4/80+SiglecF+SSChi;
  • Panel C shows representative flow cytometric images of eosinophils in pleural lavage fluid at different time points after intrapleural injection of LLC;
  • Figure D shows the proportion of white blood cells at different time points after intrathoracic injection of LLC.
  • Figure 2 shows the cytokine characteristics in the pleural lavage fluid of MPE mice.
  • A used the limma R package to perform differential expression analysis on the data, and B used two-way ANOVA for statistical analysis.
  • P ⁇ 0.05 indicates statistical difference, and n.s. indicates no statistical difference.
  • pleural lavage fluid represents pleural lavage fluid.
  • Figure C shows the transcriptional characteristics of eosinophils in pleural lavage fluid.
  • Figure 3 shows that rmCCL11 promotes eosinophil recruitment and alleviates MPE.
  • Figure A is the in vivo experimental design diagram;
  • Figure B is a representative picture of eosinophil FC;
  • Figure C is a statistical diagram of the percentage of eosinophils/total cells.
  • Figure D is a representative picture of LLC-MPE mice in vivo imaging, and the unit of bioluminescence is photons/second/cm 2 /steradian; where Eos-null represents transgenic mice lacking eosinophils.
  • FIG. 4 shows that, in MPE nude mice, reinfusion of eosinophils did not affect pleural tumor growth and pleural effusion formation.
  • Figure A is a representative FC picture of eosinophils in the pleural lavage fluid of LLC-MPE nude mice on the second day
  • Figure C is a representative picture of the in vivo imaging of LLC-MPE nude mice, and the unit of bioluminescence is photons/second/cm2/steradian
  • Figure E is a statistical chart of pleural effusion volume, and the grouping is the same as D
  • Figure F is a representative picture of in vivo
  • Figure 6 shows the phenotype analysis of immune cells in the thoracic microenvironment of WT mice and Eos-null mice after intrathoracic injection of LLC.
  • Figure A is the tSNE map of CD45+ cells in the thoracic cavity of WT and Eos-null mice;
  • Figure B is the frequency map of immune cell subsets;
  • Figure C is the tSNE map of CD3+T cells in the thoracic cavity of WT and Eos-null mice;
  • Figure D is the frequency map of T cell subsets;
  • Figure E is the expression of Ki67 and CD69 on CD3+T cells in the two groups. Data are shown as mean ⁇ standard error.
  • Figure 7 shows the effects of eosinophils on tumor cells and tumor tissues.
  • Figures A-D show the evaluation results of eosinophil killing of tumor cells.
  • Figure A shows the DAPI representative flow cytometry histograms of LLC, 4T1 and MC38 tumor cells, and tumor cells and eosinophils were co-cultured in non-contact with different ratios.
  • Figure B shows the survival rate of LLC, 4T1 and MC38 tumor cells in the non-contact co-culture system.
  • Figure C shows the gating strategy of the contact co-culture system (upper figure).
  • 7-AAD representative flow cytometry histograms of LLC, 4T1 and MC38 tumor cells in contact co-culture (lower figure).
  • Figure D shows the survival rate of LLC, 4T1 and MC38 tumor cells in the contact co-culture system.
  • Figures E-G show the pleural tumor tissues in the LLC-MPE model of WT and Eos-null mice, and the results of cleaved-caspase 3 and Ki-67 were analyzed using immunohistochemical staining.
  • Figure E shows representative IHC images of cleaved-caspase 3 and Ki-67 in pleural tumors. Scale bar: 100 ⁇ m.
  • Panel F shows the number of cleaved-caspase 3+ cells per mm2.
  • Panel G shows the number of Ki-67+ cells per mm2. Data are presented as mean ⁇ standard error.
  • the inventors unexpectedly discovered for the first time that the recombinant protein CCL11 (or the corresponding preparation) can be used to treat malignant pleural effusion (MPE).
  • MPE malignant pleural effusion
  • active ingredient of the present invention refers to the recombinant protein CCL11, a nucleic acid molecule encoding the recombinant protein CCL11 (such as mRNA, nucleic acid vector), or a combination thereof.
  • composition of the present invention refers to a composition, especially a pharmaceutical composition, containing the active ingredients of the present invention.
  • MPE is used interchangeably with "MPE complication” and refers to pleural effusion caused by malignant pleural mesothelioma originating in the pleural cavity or metastasis of tumors from other sites such as the lungs, gastrointestinal tract, and breast to the pleural cavity.
  • CCL11 is an important chemokine in the human body and a member of the CC chemokine family. CCL11 can bind to the chemokine receptor CCR3 expressed on the surface of eosinophils.
  • the recombinant CCL11 of the present invention also includes an active fragment or fusion protein of the CCL11 protein, as long as the active fragment or fusion protein retains the function of recruiting eosinophils.
  • the retained function of recruiting eosinophils refers to having ⁇ 50%, preferably ⁇ 70%, ⁇ 90% (such as 50-300%, or 70-150%) of the eosinophil recruitment function of the wild-type CCL11 protein.
  • Recombinant protein CCL11 treats and/or alleviates MPE
  • recombinant CCL11 protein can be administered to reach the chest cavity, thereby recruiting eosinophils in the chest cavity or surrounding tissues, reshaping the MPE immune microenvironment, and thereby achieving the treatment or relief of MPE complications.
  • the present invention also provides a pharmaceutical composition containing the recombinant protein CCL11, and the pharmaceutical composition can be used to treat and/or alleviate MPE.
  • the pharmaceutical composition of the present invention may further include other active ingredients, such as additional immunostimulants, anticancer drugs, or a combination thereof.
  • the pharmaceutical composition of the present invention comprises (a) a recombinant protein CCL11 or a nucleic acid molecule encoding the recombinant protein CCL11 as an active ingredient; and (b) a pharmaceutically acceptable carrier.
  • the term "effective amount” or “effective dose” refers to an amount that can produce a function or activity on humans and/or animals and can be accepted by humans and/or animals.
  • pharmaceutically acceptable ingredients are suitable for use in humans and/or mammals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), i.e., substances with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable carrier refers to a carrier for administration of a therapeutic agent, including various excipients and diluents.
  • the pharmaceutical composition of the present invention contains a safe and effective amount of the active ingredient of the present invention and a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier include (but are not limited to): saline, buffer, glucose, water, glycerol, ethanol, and combinations thereof.
  • the pharmaceutical preparation should match the mode of administration, and the dosage form of the pharmaceutical composition of the present invention is an injection, a transdermal agent, or a sustained-release agent.
  • it is prepared by conventional methods using physiological saline or an aqueous solution containing glucose and other adjuvants.
  • the pharmaceutical composition is preferably manufactured under sterile conditions.
  • the effective amount of the active ingredient of the present invention may vary with the mode of administration and the severity of the disease to be treated.
  • the selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials).
  • the factors include, but are not limited to: pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated by the patient, the patient's body weight, the patient's immune status, the route of administration, etc.
  • pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.
  • the severity of the disease to be treated by the patient the patient's body weight, the patient's immune status, the route of administration, etc.
  • the active ingredient of the present invention is administered at a dose of about 0.00001 mg-50 mg/kg animal body weight (preferably 0.0001 mg-10 mg/kg animal body weight) per day, satisfactory results can be obtained. For example, depending on the urgency of
  • representative administration methods include (but are not limited to): intrathoracic administration, such as intrathoracic injection.
  • the average daily dose for a 60 kg body weight subject is usually 1-1000 mg, preferably 5-500 mg, more preferably 10-250 mg.
  • the daily dose can be administered once, twice or multiple times, or once every 1, 2, 3, 4, 5, or 6 days.
  • the drug can also be administered according to the volume of malignant pleural effusion.
  • the pharmaceutical composition of the present invention is administered intrathoracically or intravenously, the usual daily dose is 0.1-10 mg CCL11 protein/100 mL, preferably 0.5-5 mg CCL11 protein/100 mL.
  • the daily dose can be administered once, twice or multiple times, or once every 1, 2, 3, 4, 5, or 6 days.
  • the pharmaceutically acceptable carriers of the present invention include (but are not limited to): water, saline, liposomes, lipids, peptides, cellulose, nanogels, or combinations thereof.
  • the choice of carrier should match the mode of administration, which are well known to those of ordinary skill in the art.
  • the main advantages of the present invention include:
  • the present invention proposes and verifies for the first time that intrathoracic injection of recombinant CCL11 protein can significantly and rapidly alleviate MPE, and this process is mainly achieved through eosinophils.
  • mice for constructing MPE were taken and injected with 1% sodium pentobarbital solution intraperitoneally for general anesthesia.
  • mice entered a deep anesthesia state they were fixed on the experimental table, the hair on the right chest of the mice was shaved with a shaver, and the skin was disinfected with 75% alcohol.
  • a microsyringe was used to draw 50 ⁇ L of PBS single cell suspension containing 1.5 ⁇ 10 5 tumor cells (LLC cells, MC38 cells, 4T1 cells), and an incision was made in the right anterior lateral chest wall at the level of the xiphoid process of the mouse to expose the iliac bone.
  • the single cell suspension was injected into the mouse pleural cavity through the intercostal space under direct vision. After removing the syringe, the injection point was disinfected, the muscle layer and the epidermis were sutured with surgical thread, and the mouse was placed in a warm mouse cage, named LLC-MPE model mouse.
  • the injection solution in the normal control group was 50 ⁇ L PBS solution, and other operations were the same as those in the LLC-MPE model group.
  • mice For mice, the criteria for judging MPE complications are as follows: when the volume of pleural effusion in mice is ⁇ 100 ⁇ L, the mice are considered to have MPE (fluid aspiration can be performed); when the volume of pleural effusion in mice is ⁇ 100 ⁇ L, the mice are considered to have no MPE (pleural lavage can be performed).
  • RNA sequencing of eosinophils in the pleural effusion (before administration) of LLC-MPE model mice was performed.
  • the results showed that the expression of markers such as Cxcl13, Ccl3, and Ccl4 was upregulated, and the expression of markers such as S100a9, Ear1, and Ear6 was downregulated.
  • Example 2 CCL11 promotes eosinophil recruitment and alleviates MPE
  • the present invention constructs LLC-MPE model in WT and Eos-null mice, and intrathoracic injection of mouse recombinant CCL11 (0.5 ⁇ g/mouse) begins on the 4th day, with additional injection every 2 days, and the mice are killed on the 14th day, the pleural effusion and tumor tissue of the mice are collected, and the survival period of the mice is calculated.
  • Example 3 Infusion of eosinophils into MPE nude mice does not affect pleural tumor growth and pleural effusion formation
  • Example 4 Changes of CD4+T cells and CD8+T cells in the chest cavity of MPE mice over time
  • This example further analyzes the timing of the appearance of adaptive immune cells in the MPE thoracic microenvironment.
  • the operation was the same as that of the LLC-MPE model.
  • Flow cytometry was used to detect the number of CD4 + T cells and CD8 + T cells in the chest cavity at different time points.
  • CD4+T cells and CD8+T cells gradually increased on the 7th day.
  • Example 4 the pleural lavage fluid of the LLC-MPE model of WT mice and Eos-null mice on the 7th day was obtained, and after in vitro stimulation with phorbol ester (PMA)/ionomycin (Ion)/Brefeldin A (BFA) for 4 hours, mass spectrometry flow detection was performed.
  • the acquired data was circled with Flowjo for single, living, and complete CD45+ immune cells, and the CD45+ cells were subjected to non-biased automatic dimensionality reduction clustering analysis, and 32 cell subsets were analyzed and displayed, and manual annotations were performed.
  • PMA phorbol ester
  • Ion ionomycin
  • BFA Brefeldin A
  • CD3+T cells were further circled by Flowjo for T cell subset analysis.
  • CD3+T cells were subjected to non-biased automatic dimensionality reduction cluster analysis, showing the distribution of 8 subsets.
  • the immune cell phenotypes in the thoracic microenvironment of WT mice and Eos-null mice after intrathoracic injection of LLC were analyzed. The results showed that the absence of eosinophils would affect the immune cell composition and T cell function of the thoracic microenvironment of MPE mice, thereby promoting the occurrence and development of MPE.
  • eosinophils were co-cultured with three different tumor cells in different proportions to observe the effects of eosinophils on tumor cells and tumor tissues.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Hematology (AREA)
  • Diabetes (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention divulgue l'utilisation d'une protéine CCL11 recombinante dans le traitement d'effusions pleurales malignes. Spécifiquement, la présente invention concerne une méthode de traitement et/ou de soulagement des effusions pleurales malignes (MPE) au moyen d'une injection intrapleurale de la protéine CCL11 recombinante. La présente invention a découvert dans un premier temps que CCL11 peut soulager les MPE au moyen de l'augmentation des granulocytes éosinophiles dans la cavité thoracique. Un modèle animal de MPE est construit, et l'injection intrapleurale de la protéine CCL11 recombinante augmente remarquablement les granulocytes éosinophiles dans la cavité thoracique, et parallèlement, soulage évidemment les MPE.
PCT/CN2024/140718 2023-12-31 2024-12-19 Utilisation d'une protéine ccl11 recombinante dans le traitement d'effusions pleurales malignes Pending WO2025140008A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202311869406.9A CN117815369A (zh) 2023-12-31 2023-12-31 重组蛋白ccl11在治疗恶性胸腔积液中的应用
CN202311869406.9 2023-12-31

Publications (1)

Publication Number Publication Date
WO2025140008A1 true WO2025140008A1 (fr) 2025-07-03

Family

ID=90509347

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2024/140718 Pending WO2025140008A1 (fr) 2023-12-31 2024-12-19 Utilisation d'une protéine ccl11 recombinante dans le traitement d'effusions pleurales malignes

Country Status (2)

Country Link
CN (1) CN117815369A (fr)
WO (1) WO2025140008A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117815369A (zh) * 2023-12-31 2024-04-05 浙江大学 重组蛋白ccl11在治疗恶性胸腔积液中的应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023169686A1 (fr) * 2022-03-10 2023-09-14 Buzzard Pharmaceuticals AB Inhibiteur du récepteur de l'interleukine-1 de type 1 destiné à être utilisé dans le traitement du cancer
CN117815369A (zh) * 2023-12-31 2024-04-05 浙江大学 重组蛋白ccl11在治疗恶性胸腔积液中的应用

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023169686A1 (fr) * 2022-03-10 2023-09-14 Buzzard Pharmaceuticals AB Inhibiteur du récepteur de l'interleukine-1 de type 1 destiné à être utilisé dans le traitement du cancer
CN117815369A (zh) * 2023-12-31 2024-04-05 浙江大学 重组蛋白ccl11在治疗恶性胸腔积液中的应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
GHAFFARI SASAN, REZAEI NIMA: "Eosinophils in the tumor microenvironment: implications for cancer immunotherapy", JOURNAL OF TRANSLATIONAL MEDICINE, BIOMED CENTRAL, vol. 21, no. 1, XP093331222, ISSN: 1479-5876, DOI: 10.1186/s12967-023-04418-7 *
YANG JINMING, HAWKINS ORIANA E., BARHAM WHITNEY, GILCHUK PAVLO, BOOTHBY MARK, AYERS GREGORY D., JOYCE SEBASTIAN, KARIN MICHAEL, YU: "Myeloid IKKβ Promotes Antitumor Immunity by Modulating CCL11 and the Innate Immune Response", CANCER RESEARCH, UNIVERSITY OF CHICAGO PRESS, vol. 74, no. 24, 15 December 2014 (2014-12-15), pages 7274 - 7284, XP093331231, ISSN: 0008-5472, DOI: 10.1158/0008-5472.CAN-14-1091 *
ZHANG MIN, LI FEI, SHEN HUAHAO: "Research on the role of eosinophils in the pathogenesis of malignant pleural effusion", 2020 (42ND) ZHEJIANG PROVINCIAL MEDICAL ASSOCIATION RESPIRATORY DISEASES ACADEMIC CONFERENCE, 27 August 2020 (2020-08-27), pages 8 - 8, XP093331220, DOI: 10.26914/c.cnkihy.2020.071794 *
ZHANG MIN, XIA LIXIA, PENG WENBEI, XIE GUOGANG, LI FEI, ZHANG CHAO, SYEDA MADIHA ZAHRA, HU YUE, LAN FEN, YAN FUGUI, JIN ZHANGCHU, : "CCL11/CCR3-dependent eosinophilia alleviates malignant pleural effusions and improves prognosis", NPJ PRECISION ONCOLOGY, NATURE PUBLISHING GROUP UK, vol. 8, no. 1, XP093331207, ISSN: 2397-768X, DOI: 10.1038/s41698-024-00608-8 *

Also Published As

Publication number Publication date
CN117815369A (zh) 2024-04-05

Similar Documents

Publication Publication Date Title
Xu et al. Mesenchymal stem cell‐derived extracellular vesicles with high PD‐L1 expression for autoimmune diseases treatment
Yang et al. Deficiency of GATA3-positive macrophages improves cardiac function following myocardial infarction or pressure overload hypertrophy
Xue et al. The role of dendritic cells regulated by HMGB1/TLR4 signalling pathway in myocardial ischaemia reperfusion injury
CN114340667A (zh) 通过向癌细胞施加交变电场和给予检查点抑制剂来降低癌细胞生存能力的方法
JP2022095884A (ja) 単離ミトコンドリアを含む関節リウマチの予防または治療のための医薬組成物
Mou et al. Macrophage‐targeted delivery of siRNA to silence Mecp2 gene expression attenuates pulmonary fibrosis
CN106167789B (zh) 低氧处理的间充质干细胞及其应用
Wu et al. Anti-ST2 nanoparticle alleviates lung inflammation by targeting ILC2s-CD4+ T response
WO2025140008A1 (fr) Utilisation d'une protéine ccl11 recombinante dans le traitement d'effusions pleurales malignes
US20240287136A1 (en) Pharmaceutical composition and the use thereof in the treatment of autoimmune diseases
CN114099481A (zh) 雾化吸入型糖皮质激素纳米药物及其制备方法和应用
US11622964B2 (en) Method for destroying cellular mechanical homeostasis and promoting regeneration and repair of tissues and organs, and use thereof
US20220362181A1 (en) Methods of treating acute respiratory distress syndrome using colchicine
JP6549304B2 (ja) ナノ粒子−硝子体基盤タンパク質複合体を有効成分として含む血管新生抑制用組成物およびその用途
US20130072844A1 (en) Use of entrained neutrophils to treat metastatic and micrometastatic disease in at risk patients
CN112168966A (zh) 干预nlrp3炎症小体在制备治疗新型冠状病毒肺炎的药物中的应用
CN114099545B (zh) 外泌体在胸主动脉夹层/动脉瘤中的作用
TWI721352B (zh) 含有ｇｍ３神經節苷酯作為免疫調節劑之奈米粒子
KR101336386B1 (ko) 림프관 생성 유도제
JP2018517406A (ja) 腫瘍に対して直接阻害作用を有するインターフェロンの同定方法、及びその使用
WO2021004229A1 (fr) Nouvelle application de l'inhibiteur du récepteur de chimiokine ccr6 dans la prévention de la récurrence du psoriasis
CN115300507B (zh) I-brd9作为arih1激动剂的用途
US20250223555A1 (en) Reprogramming of immune-enhancing neutrophils by subclinical low-dose endotoxin for the treatment of cancer
Zhao et al. Innate immune checkpoint SIRPα/CD47 blockade ameliorates silica-induced pulmonary fibrosis by modulating macrophage immunity
CN119607021A (zh) 一种外泌体在制备治疗脓毒症急性肺损伤药物中的应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24910937

Country of ref document: EP

Kind code of ref document: A1