58 results about "Hypoxic tumor" patented technology

The invention relates to a strontium peroxide nano-material, which is characterized in that a strontium source, alkali and hydrogen peroxide are used as raw materials, a mixed solution of an organic solvent and water is used as a reaction solvent, the strontium peroxide nano-material is prepared under a template or template-free condition, and the strontium peroxide nano-material can quickly react with water to generate hydroxyl, hydrogen peroxide and oxygen, hydrogen peroxide can achieve a tumor cell killing effect, hydroxyl can neutralize an acidic tumor microenvironment and alkalify tumor cells, and oxygen can relieve a hypoxic tumor microenvironment. In addition, the strontium peroxide nano material can induce macrophages to be polarized to an anti-tumor M1 type by regulating and controlling glycometabolism of the macrophages, and remodeling an intra-tumor lactic acid metabolism crosstalk mode. The strontium peroxide nano-material or the modified material and the preparation thereof can be directly injected into a tumor or near the tumor under ultrasonic guidance to realize in-situ treatment, or can be delivered to a tumor supply blood vessel through a catheter to realize embolism treatment, so that the strontium peroxide nano-material or the modified material and the preparation thereof have a certain application prospect in the field of tumor treatment.

A preparation method of the metal organic coordination network comprises the following steps: respectively dissolving copper acetate and polyphenol in deionized water or dimethyl sulfoxide to form a copper acetate solution and a polyphenol solution, then dropwise adding the copper acetate solution into the polyphenol solution, introducing nitrogen, and carrying out a heating reaction to obtain a reaction mixture; centrifuging the reaction mixture to obtain a precipitation product metal organic coordination network; the metal organic coordination network is applied to preparation of a material for targeted killing of hypoxic tumor cells. The metal organic coordination network disclosed by the invention is good in dispersity, high in biocompatibility and good in hypoxic tumor cell selective toxicity. After being degraded by glutathione, the glutathione derivative has good peroxidase activity, can catalyze hydrogen peroxide to generate hydroxyl free radicals with stronger cytotoxicity, and can be used for converting tumors into oxidative phosphorylation metabolic pathways by inhibiting hypoxic tumor glycolysis and remodeling hypoxic tumor metabolism, promoting tumor cells to generate ROS (reactive oxygen species), and further improving the activity of glutathione. And the sensitivity of hypoxic tumors to copper death is enhanced.

The invention discloses a CAIX targeted degradation agent. The CAIX targeted degradation agent comprises a CAIX target protein ligand, an integrin ligand and a connector, the CAIX target protein ligand and the integrin ligand are connected through the connecting body. As the carbonic anhydrase IX is enriched in the tumor region, the CAIX targeted degradation agent can specifically target the tumor region; the compound can effectively induce degradation of CAIX and inhibit survival of tumor cells under the hypoxia condition, has the functions of targeting a tumor area and degrading carbonic anhydrase IX by utilizing integrin and lysosome, further changes hypoxia and subacid environments of the internal environment of a tumor patient, and plays a role in inhibiting survival of hypoxia tumor cells; the tumor targeting property is high, the toxic and side effects on normal cells are relatively small, and a relatively good effect can be obtained by using a relatively small dosage.

The invention specifically discloses a beta-diketone iridium complex as well as a preparation method and application thereof, and relates to the technical field of biological medicines. The beta-diketone iridium complex provided by the invention has high phototoxicity and low dark toxicity, can be applied to tumor cells as effective PSs of PDT, and is specifically delivered to the tumor cells by taking a traditional Chinese medicine monomer tripterine as a supplement in combination with a nano-targeting delivery technology for photodynamic / immune synergistic treatment of hypoxic tumors.

The invention discloses an asymmetric nanocomposite material of titanium dioxide-gold nanorods, a preparation method and application thereof, wherein the nanocomposite material has an asymmetric structure with one end of the gold nanorod as the growth point of titanium dioxide nanometers, wherein the titanium dioxide is directly in contact with one end of the gold nanorod, wherein the aspect ratio of the gold nanorod is about 2.5-3.5, and the thickness of the directly contacted titanium dioxide is 7-20 nanometers, and the nanocomposite particles have good tumor penetration due to their small size, and are absorbed in the near-infrared light region of 650-900 nanometers with strong penetration, and are surface-modified with PEG and fluorescent molecules-PEG to increase biocompatibility. The nanocomposite particles can be used for suitable photothermal and good photodynamic synergistic therapy of hypoxic tumors under near-infrared illumination; the synthesis method of the nanocomposite particles has the advantages of convenient operation, cheap raw materials, simple separation, etc.

The invention belongs to the field of medicinal chemistry, and particularly discloses a preparation method and application of a nitroimidazole derivative serving as an anti-tumor radiosensitizer. According to the invention, nitroimidazole is used as a lead compound, and a framework of nitroimidazole is modified through a series of reactions to obtain the novel nitroimidazole derivative. The compound can target a tumor hypoxia region, is reduced by a specific enzyme highly expressed in a hypoxia tumor tissue, and is combined with nucleophilic biological macromolecular free radicals generated by radiation to accelerate cancer cell death. As an anti-tumor radiosensitizer, the compound disclosed by the invention is expected to improve the tumor cell selectivity of the compound and reduce toxic and side effects, and is used for subsequent anti-tumor activity research.

This invention provides a biomarker for detecting immunotherapy resistance in gastric cancer, its application, a model, and a method for using it. This invention demonstrates the significant impact of the hypoxic tumor microenvironment on immunotherapy resistance in gastric cancer, highlighting the crucial roles of FOXO3a and PD-L1 expression in mediating immune escape mechanisms. By employing an innovative microfluidic tumor chip model, the in vivo hypoxic tumor environment was successfully simulated, revealing how hypoxia-induced upregulation of FOXO3a leads to enhanced PD-L1 expression, thereby resulting in immunotherapy resistance.

The invention provides an acoustic-magnetic dual-mode diagnosis and treatment agent as well as a preparation method and application thereof, and belongs to the technical field of preparation of tumor diagnosis and treatment agents. The acoustic-magnetic dual-mode diagnosis and treatment agent is surface-modified magnetic nanoparticles which contain nano balloon genes and are subjected to Escherichia coli induced expression; the nucleotide sequence of the nano-balloon gene is a sequence shown as NCBI: KC601845.1, escherichia coli in the acoustic-magnetic dual-mode diagnosis and treatment agent as a diagnosis and treatment agent carrier has an active targeting property on a tumor hypoxic region, and bacteria contain a nano-balloon and have an acoustic response characteristic; the escherichia coli surface modified magnetic nanoparticles have a magnetic response characteristic. In acoustic-magnetic dual-mode imaging, firstly, on a real-time ultrasonic contrast monitoring image, a nano air bag expressed in bacteria can generate an enhanced tumor ultrasonic echo signal; secondly, under the action of an external changing magnetic field, the magnetic nanoparticles on the surfaces of the bacteria can generate an enhanced tumor magnetic vibration imaging effect, so that more efficient tumor diagnosis and / or treatment can be realized.

The invention discloses a method for preparing a Prussian blue / chitosan / sodium alginate composite hydrogel, which comprises the steps of in-situ polymerization of Prussian blue by chitosan and the step of compounding sodium alginate. The step of in-situ polymerization of Prussian blue by chitosan is to first modify chitosan by carboxyethylation, and then polymerize with Prussian blue. In the present invention, the combined treatment of Prussian blue loaded with doxorubicin hydrogel assisted by photothermal therapy can be injected non-invasively into the lesion while maintaining the drug concentration in the tumor site for a long period, improving the bioavailability of the drug, increasing the degree of T cell infiltration in the tumor site of patients with low immunogenic tumors, thereby improving the efficacy of the drug, and minimizing toxic side effects, decomposing endogenous H2O2 into O2, alleviating the hypoxic tumor microenvironment and releasing the anticancer drug DOX at the same time, for effective photothermal chemotherapy of breast cancer.

The invention relates to a preparation method of a heavy-atom-free photosensitizer based on a BODIPY dye, the photosensitizer comprises a thiocarbamate light cage, not only can generate singlet oxygen (1O2) under red light irradiation, but also can release carbonyl sulfide (COS), and generates hydrogen sulfide (H2S) under physiological conditions through carbonic anhydrase catalysis. By reasonably adjusting the molecular structure, the singlet oxygen quantum yield and hydrogen sulfide release efficiency of the photosensitizer are remarkably improved. The method is the first case which is reported at present and can simultaneously activate and release the organic optical functional micromolecules of H2S and 1O2 through a single beam of red light. The strategy effectively solves the problem of dependence on oxygen in type II photodynamic therapy (PDT), and further effectively improves the effect of PDT hypoxic tumors. In in-vitro and in-vivo experiments, hydrogen sulfide gas and photodynamic synergic treatment trigger pyroptosis of cells, promote maturation of dendritic cells and stimulate antitumor immune response. Therefore, the invention provides a synergistic treatment strategy combining gas, photodynamic and immunotherapy, and has a remarkable effect in the aspect of hypoxic tumor treatment. The photosensitizer material disclosed by the invention is simple to prepare and low in cost, has a wide application prospect, and particularly provides a solution with huge potential for cancer treatment in the field of biomedicine.

The invention belongs to the technical field of medicine preparation, and particularly relates to a ferroptosis agonist, an anoxia response type nanometer co-delivery system and application thereof. The novel ferroptosis agonist RSL3-ClAc is synthesized by introducing a chloracetyl fragment into an RSL3 molecule, the binding capacity of the novel ferroptosis agonist RSL3-ClAc with GPX4 is remarkably enhanced, and then the ferroptosis induction efficiency is improved. And meanwhile, the RSL3-ClAc and the hypoxia response type photosensitizer TCy5-NO2 are subjected to self-assembly, so that the nano drug delivery system is constructed. According to the nano drug delivery system, the EPR effect and the hypoxia response characteristic of the nano carrier are utilized, the tumor barrier can be effectively broken through, and the focus can be accurately positioned. In a tumor hypoxic microenvironment, the activation of a Try5-NO2 photosensitizer is triggered by nitroreductase (NTR), and RSL3-ClAc is synchronously released to enhance the ferroptosis induction effect.

The application relates to a composite nanogel and a preparation method and application thereof, in particular to the technical field of biomedical materials. The composite nanogel comprises a nanogel framework and tirapazamine and folic acid loaded on the nanogel framework, wherein raw material components of the nanogel framework comprise a temperature-sensitive monomer and a monomer containing a catechol group. The composite nanogel provided by the application can solve the problems that poor blood vessel adhesion of traditional embolic agents easily leads to embolism recanalization, simple physical embolism is difficult to completely remove hypoxic tumor cells, and existing treatment strategies cannot effectively reverse the immune suppression microenvironment after embolism and activate the systemic anti-tumor immune response.

This application provides methods of inhibiting an inflammation- or hypoxia-induced death of a cell involving inhibiting an RNF213 signaling pathway. The application further provides methods of preventing or treating various diseases and disorders in a subject involving administering to the subject an inhibitor of expression of RNF213 protein or an inhibitor of one or more functions of the RNF213 protein. The application still further provides methods for enhancing the effectiveness of a cancer treatment in a subject where the cancer involves a hypoxic tumor microenvironment involving co-administering the cancer treatment with an inhibitor or activator of different functions of the RNF213 protein. The application also provides methods of eliminating drug-resistant tumor cells by co-administering an inhibitor of the expression of RNF213 protein or by inhibiting or activating specific functions of the RNF213 protein.

The application discloses a kind of lipid droplet targeted NIR-I photothermal agent and its application in hypoxic tumor treatment, belong to the technical field of photothermal therapy.The NIR-I photothermal agent of the application is a kind of organic photothermal material based on donor-acceptor structure, with strong electron-deficient and rigid planar structure dithiophene pyrrolone sub benzodifuran diketone (BTPDBDF) as acceptor unit, respectively with triphenylamine and tetraphenyl ethylene as donor unit, with strong intramolecular charge transfer, absorption spectrum reaches near infrared region one.Simultaneously, the material is prepared into water-soluble nanoparticles by nano precipitation method, and the biocompatibility and targeting are improved.The nano preparation has the ability of specific targeting cell lipid droplet, and shows significant heating performance and high photothermal conversion efficiency under 808 nm laser irradiation, and shows good near-infrared light capturing capacity, so as to facilitate effective photothermal therapy under the condition of tumor hypoxia.

This invention discloses a class of enzyme-activated prodrug compounds having the following general structural formula I: This invention discloses a class of enzyme-sensitive chlorambucil analog prodrugs. These prodrugs exhibit high selectivity for hypoxic tumors and can be effectively activated within hypoxic tumor cells. Upon specific activation, the prodrug releases free chemotherapeutic drug chlorambucil, simultaneously releasing the photosensitizer cyanine dye, achieving an "OFF-ON" photosensitivity conversion, and generating a large number of reactive oxygen species, thus achieving the unification and synergy of photodynamic therapy and activatable chemotherapy.

The invention discloses a ruthenium complex of targeted carbonic anhydrase as well as a preparation method and application of the ruthenium complex. The complex takes ruthenium (Ru < 2 + >) as a central metal ion, is formed by coordinating a bidentate ligand (An-en) containing an anthracene structure, a pyridine derivative ligand (N-ABS) containing a benzenesulfonamide group and a ruthenium precursor, and has the capability of accurately targeting tumor cell carbonic anhydrase IX (CAIX); the complex is low in dark toxicity and excellent in curative effect in an anoxic environment, the defects that a traditional ruthenium complex is high in dark toxicity and poor in anoxic curative effect can be effectively overcome, and an efficient and low-toxicity new strategy is provided for anoxic tumor treatment.

This invention belongs to the field of pharmaceutical chemistry technology, specifically relating to a near-infrared II luminescent iridium(III) complex photosensitizer, its preparation method, and its application. The near-infrared II luminescent iridium(III) complex photosensitizer disclosed in this invention generates singlet oxygen (…) under 550 nm illumination. 1 O2), hydroxyl radicals (•OH), superoxide anions (O2) •‑ We evaluated the antitumor activity of near-infrared luminescent iridium complexes using mouse breast cancer cells (4T1). Under unilluminated conditions, the near-infrared II luminescent iridium(III) complex photosensitizers showed no significant cytotoxicity to 4T1 cells, while under illumination, they exhibited good phototoxicity to 4T1 cells, with an IC50 value of [missing value]. 50 The concentration is 15.92 μM. We have invented a near-infrared II luminescent iridium(III) complex photosensitizer, which has excellent photoimmunodynamic therapeutic effects and will have important application prospects in the treatment of hypoxic tumors.

The invention discloses a polygonal anthracene functionalized BODIPY supramolecular metal ring compound as well as a preparation method and application thereof, and belongs to the technical field of supramolecular materials, the polygonal anthracene functionalized BODIPY supramolecular metal ring compound is obtained by self-assembly reaction of an anthracene functionalized BODIPY photosensitizer with a specific structure and a platinum compound; by selecting platinum compounds with different coordination included angles, triangular, quadrilateral and hexagonal supramolecular metal ring compounds are respectively prepared. Furthermore, amphiphilic polymer F127 is used for wrapping the supramolecular metal ring compound to form water-soluble nanoparticles, so that phototherapy and chemotherapy combined therapy of hypoxic tumors can be realized, and the amphiphilic polymer F127 has a wide application prospect in the aspects of biological imaging and tumor therapy.

The present application relates to the technical field of nanobiomedicine, and particularly relates to a porphyrin oxygen-carrying polymer sonosensitizer, a preparation method and application thereof; the porphyrin oxygen-carrying polymer sonosensitizer takes porphyrin as a main body structure and takes polyheptafluorobutyramide as a polymer side chain. The polyheptafluorobutyramide contained in the porphyrin oxygen-carrying polymer sonosensitizer can carry oxygen, and under the activation of ultrasound, the perfluoroalkyl-modified polymer can release oxygen explosively, the perfluoroalkyl-modified polymer triggered by visible light increases oxygen supply to relieve tumor hypoxia, reverses a low-oxygen microenvironment, enhances the generation of reactive oxygen species in the SDT process, further improves the accumulation efficiency at a tumor site, improves the effect of SDT in treating tumors, and becomes an effective method for enhancing the chemical sonodynamic therapy of hypoxic tumors.

The invention relates to the field of biochemical materials and organic photoelectric information materials, and discloses a near-infrared nitric oxide probe, when the near-infrared probe responds to nitric oxide, a novel phototherapy reagent can be generated, and under the irradiation condition of 550-730 nm near-infrared light, the near-infrared probe can be used for detecting the near-infrared nitric oxide. The phototherapy reagent shows enhanced active oxygen generation capability and performance of continuously releasing nitric oxide, shows remarkably enhanced photoacoustic signals and photo-thermal performance in the nitric oxide releasing process, realizes combined treatment of hypoxic tumors, including hypoxic tumor photo-thermal treatment, I-type photodynamic treatment and nitric oxide mediated gas therapy, and also has the advantages that the phototherapy reagent can be used for treating hypoxic tumors, and the phototherapy reagent can be used for treating hypoxic tumors. After nitric oxide is released, the system can recover to obtain the initial nitric oxide probe, so that the probe has the characteristics of reversible combination and nitric oxide release.

The present invention belongs to the field of biomedical technology, and discloses an organic photosensitive nanomaterial, a preparation method thereof and an application thereof. The organic photosensitive nanomaterial of the present invention has a core-shell structure; the shell structure includes an amphiphilic polymer, and the core structure includes an organic conjugate and an electron sacrificial agent; the preparation method of the organic photosensitive nanomaterial of the present invention includes the following steps: self-assembling the organic conjugate, the electron sacrificial agent and the amphiphilic polymer. Due to its unique structural design and excellent GSH consumption performance, the organic photosensitive nanomaterial of the present invention can consume endogenous GSH in tumor cells, reduce its loss of ROS, and further enhance the oxidative stress in tumor cells; and can jointly regulate the level of GSH in tumor cells and increase the utilization rate of ROS in hypoxic tumors, and is used as a drug regimen for realizing GSH consumption to promote the treatment effect of hypoxic tumors in PDT.

The invention relates to the technical field of biological medicine, in particular to a sensitizer for tumor radiotherapy and application. The sensitizer includes a hydrate of a cobalt-containing polyoxometallate. The sensitizer for tumor radiotherapy comprises the hydrate of cobalt-containing polyoxometallate, and the cobalt-containing polyoxometallate can be metabolized in vivo, has no side effect and is high in cost performance. Other high-atomic-number metals (such as tungsten) in the cobalt-containing polyoxometallate can enhance the radiation dose of local tumors, and cobalt metal can be released from the sensitizer through X-rays to induce pyroptosis of hypoxic tumor cells, so that the sensitivity of hypoxic irradiation tolerant tumor cells to radiotherapy is improved.

The invention belongs to the technical field of biological medicine, and particularly relates to preparation of a hypoxia response albumin supramolecular nano drug delivery system and application of the hypoxia response albumin supramolecular nano drug delivery system in hypoxia tumor treatment. The nano drug delivery system comprises an albumin supramolecular nano carrier taking an azo calix [4] arene derivative as a drug carrying main body, a mitochondrial targeting photosensitizer and an autophagy inhibitor. The system is prepared through a light click chemical reaction between double bonds of azo calixarene and albumin subjected to reduction treatment, and has the hypoxia response characteristic. Meanwhile, the nano drug delivery system can initiate an active oxygen storm effect and inhibit the growth of hypoxic tumors in a hypoxic environment and under external stimulation of red light, and has a fluorescence imaging function. In addition, the novel drug delivery system is mild in preparation process, can load multiple drug combinations at the same time, has a diagnosis and treatment integrated function, and has a wide application prospect in the field of hypoxia tumor treatment.

The present invention relates to a method using low intensity pulsed ultrasound (LIPUS) in order to improve drug delivery efficiency in a hypoxic tumor microenvironment (TME) of cholangiocarcinoma (CCA), and more specifically, to a method for confirming the drug permeability and cancer cell apoptosis of cholangiocarcinoma through the irradiation of low intensity pulsed ultrasound (LIPUS) under a tumor microenvironment, and a combination regimen, of gemcitabine and cisplatin with low-intensity pulsed ultrasound, for inhibiting cholangiocarcinoma growth. To this end, provided is the method for confirming the drug permeability and cancer cell apoptosis of cholangiocarcinoma through the irradiation of low intensity pulsed ultrasound (LIPUS) under a tumor microenvironment, the method comprising: a step (S100) for administering a drug and a fluorescent material to cholangiocarcinoma (CCA) under a tumor microenvironment; a step (S120) for allowing a predetermined time to elapse; a step (S140) for irradiating the cholangiocarcinoma (CCA) low intensity pulsed ultrasound (LIPUS); a step (S160) for acquiring a 3D image by performing tissue-clearing on the cholangiocarcinoma (CCA); and a step (S180) for confirming the in vivo drug permeability of a drug and a fluorescent material from the 3D image.

The invention relates to composite nanogel as well as a preparation method and application thereof, in particular to the technical field of biomedical materials. The composite nanogel comprises a nanogel skeleton, tirapazamine and folic acid, the tirapazamine and the folic acid are loaded on the nanogel skeleton, and raw material components of the nanogel skeleton comprise a temperature-sensitive monomer and a monomer containing a catechol group. The composite nanogel provided by the invention can solve the problems that a traditional embolism agent is poor in blood vessel adhesion and easy to cause embolism recovery, hypoxia tumor cells are difficult to thoroughly remove by pure physical embolism, and an immunosuppressive microenvironment after embolism cannot be effectively reversed and systemic anti-tumor immune response cannot be activated by an existing treatment strategy.

The invention relates to the technical field of tumor diagnosis, in particular to a tumor hypoxic PET developer as well as a precursor compound, application and a preparation method thereof. The precursor compound has a structure of any one of formulas I to III. The tumor hypoxia PET developer provided by the invention is used for diagnosing hypoxia tumors, and has the advantages of strong stability, good water solubility and high safety.

The present invention discloses a fluorinated zinc phthalocyanine derivative and its preparation method and application. The fluorinated zinc phthalocyanine derivative is obtained by chemically covalently linking trithiocarbonate to the four amino groups of tetraaminoperfluorophthalocyanine zinc. The fluorinated zinc phthalocyanine derivative can be used as a star-shaped RAFT chain transfer agent to induce hydrophobic methacrylate monomers and / or hydrophilic acrylate monomers to prepare four-arm star-shaped random copolymers through near-infrared light-controlled reversible addition-fragmentation chain transfer polymerization. The star-shaped polymer prepared by the above method has a narrow molecular weight distribution and can form polymer micelles with uniform hydrodynamic diameter in an aqueous solution below its critical aggregation concentration. In addition, the above polymer micelles have a high singlet state yield and good oxygen carrying capacity, and can be used for two-photon photodynamic therapy of hypoxic tumors.

The invention belongs to the technical field of biological medicines, and discloses an organic photosensitive nano material as well as a preparation method and application thereof. The organic photosensitive nano material is of a shell-core structure; the shell structure comprises an amphiphilic polymer, and the core structure comprises an organic conjugate and an electron sacrificial agent; the preparation method of the organic photosensitive nano-material comprises the following steps: self-assembling the organic conjugate, the electron sacrificial agent and the amphiphilic polymer to obtain the organic photosensitive nano-material. Due to the unique structural design and excellent GSH consumption performance of the organic photosensitive nano material, endogenous GSH in tumor cells can be consumed, loss of ROS by the endogenous GSH is reduced, and oxidative stress in the tumor cells is further enhanced; the level of GSH in tumor cells can be jointly regulated and controlled, the utilization rate of ROS in hypoxic tumors is increased, and as a medication scheme for achieving GSH consumption, the compound is applied to PDT to promote the treatment effect of the hypoxic tumors.

The invention discloses a tumor treatment system based on an ultrasonic fatigue effect, which directly damages a cancer cell membrane structure and destroys the integrity of the cancer cell membrane structure through the continuous action of low-intensity ultrasonic energy, specifically targets a disordered tumor extracellular matrix by utilizing the ultrasonic fatigue effect, and simultaneously causes mechanical damage to tumor blood vessels. The tumor core hypoxia caused by nutrition delivery is blocked, so that cell apoptosis is induced. And the multi-layer mechanical-biological effect jointly inhibits tumor growth. The tumor treatment system is combined with temozolomide for use, can synergistically amplify the anti-tumor effect, and is suitable for hypoxic tumors tolerated by a traditional therapy. According to the treatment system, a non-invasive and controllable tumor treatment mode is achieved through accurate regulation and control of energy, and a new thought is provided for tumor treatment.

This invention relates to the field of cell culture technology, and discloses a method for culturing NK cells and its applications. When NK cells enter a hypoxic environment, not only do their cytotoxic activity and proliferative capacity rapidly decline, but they also experience metabolic disorders such as impaired mitochondrial function, insufficient ATP production, and abnormal accumulation of reactive oxygen species (ROS), leading to functional failure and limiting their application in tumor therapy. Existing technologies mainly rely on constant or single-stage hypoxia induction, failing to improve the insufficient hypoxia adaptation of NK cells at the metabolic level. Therefore, this invention provides a method that uses repeated acclimatization culture with gradually decreasing oxygen concentration to allow NK cells to gradually adapt to the metabolic demands of a hypoxic environment. NK cells cultured using this method exhibit a more stable metabolic state and superior cytotoxic activity in a hypoxic environment, significantly enhancing their anti-tumor efficacy and solving the problem of metabolic imbalance and rapid functional loss of NK cells in the hypoxic tumor microenvironment in existing technologies.