WO2017079881A1 - Procédé permettant d'améliorer la capacité de tuer des cellules anormales et composition pharmaceutique - Google Patents

Procédé permettant d'améliorer la capacité de tuer des cellules anormales et composition pharmaceutique Download PDF

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WO2017079881A1
WO2017079881A1 PCT/CN2015/094143 CN2015094143W WO2017079881A1 WO 2017079881 A1 WO2017079881 A1 WO 2017079881A1 CN 2015094143 W CN2015094143 W CN 2015094143W WO 2017079881 A1 WO2017079881 A1 WO 2017079881A1
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cells
natural killer
killer cells
cell
highly active
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PCT/CN2015/094143
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张明杰
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张明杰
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Priority to CN201580001069.9A priority Critical patent/CN107109363A/zh
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum

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  • the present invention relates to the field of biomedical, and in particular, to a method and a pharmaceutical composition for enhancing the lethality of abnormal cells.
  • Natural killer (NK) cells are important immune cells of the body, not only related to anti-tumor, anti-viral infection and immune regulation, but also participate in hypersensitivity and autoimmune diseases in some cases.
  • NK cells The role of NK cells is broad-spectrum and non-specific; for specific tumor cells, the targeting is not strong; like rockets, some shells may not hit the target and are wasted.
  • Monoclonal (monoclonal antibody) targeted drugs including but not limited to rituximab for the treatment of leukemia and Herceptin for the treatment of breast cancer, have been used clinically for many years and have achieved good results.
  • NK cells ie, antibody dependent cytotoxicity (ADCC).
  • ADCC antibody dependent cytotoxicity
  • cancer patients' own NK cell activity is often low, coupled with conventional treatment such as chemotherapy or radiotherapy, NK cell activity is usually severely damaged, which naturally limits the efficacy of monoclonal antibody targeting drugs.
  • the present invention is directed to solving one of the above technical problems at least to some extent or to provide a commercial choice.
  • a method of treating a patient comprising the step of administering a combination of NK cells and an antibody drug, the NK cells comprising highly active NK (HANK) cells, the obtaining of the HANK cells comprising: utilizing cells with cytokines
  • HANK highly active NK
  • HANK cells were obtained by the inventors by activating in vitro culture to activate NK cells derived from the body. It should be noted that the so-called HANK cells are not ordinary in vivo NK cells. Ordinary NK cells are not only small in number, but are usually in a state of inhibition, and the activity of killing abnormal cells is low, for example, the activity of killing cancer cells or virus-infected cells is low.
  • the present invention does not limit the antibody drugs to be administered in combination, and may be a monoclonal antibody targeted drug currently available on the market, or may be a newly developed monoclonal antibody targeted drug in the future.
  • the cytokine carried by the cell vacant shell may be a cytokine which is naturally expressed on the surface of the cell itself, or may be expressed by a genetic engineering method, for example, by transient transfection or stable expression. On the cell surface, it can also be a cytokine that adsorbs or crosslinks on the cell surface.
  • the cells for preparing the cell vesicle may be primary cells such as peripheral blood mononuclear cells (PBMC), or may be passage cells such as K562 cells.
  • PBMC peripheral blood mononuclear cells
  • the combination of NK cells containing the HANK cells and the antibody drug can enhance the therapeutic effects of the NK cells and the antibody drugs, and the specific expressions include: (1) the antibody drug imparts specificity to the HANK cell. Sex, such as in combination with rituximab, allows HANK cells to kill leukemia cells, and combination with Herceptin monoclonal antibody allows HANK cells to kill breast cancer cells; (2) HANK cells can be monoclonal antibodies Provide enough active factors to effectively kill cancer cells, such as a large number of granzymes, perforin and so on.
  • the method of this aspect of the invention may also have at least one of the following additional technical features:
  • the so-called patient has at least one of a cancer, a viral infection, and an immune disease.
  • the so-called patient is a cancer patient and the so-called antibody drug is a monoclonal antibody-targeted drug.
  • Monoclonal targeting drugs confer specificity to HANK cells.
  • HANK cells can kill leukemia cells.
  • Herceptin monoclonal antibody HANK cells can kill breast cancer cells; HANK The cells provide the monoclonal antibody with enough active factors to effectively kill cancer cells, such as a large amount of granzymes, perforin and the like.
  • the combined use of HANK cells and monoclonal antibody-targeted drugs can increase the efficacy of each other to better treat cancer.
  • the antibody drug is a monoclonal antibody or a polyclonal antibody.
  • the two antigen binding sites on one antibody molecule may be the same or different, ie, bispecific antibodies, located at the ends of the arms, called antigen-binding fragments (Fab).
  • Fab antigen-binding fragments
  • the so-called antibody drug is a complete monoclonal antibody drug.
  • the ratio of HANK cells to antibody drugs in the NK cells administered in combination is 2 ⁇ 10 5 to 5 ⁇ 10 5 : 1 (pieces / ⁇ g).
  • the doctor or the drug instruction will give the amount according to the patient's condition.
  • the in vitro test and the animal test show that the combined administration of the ratio of HANK cells at the time of administration can significantly enhance the effect of treating cancer.
  • the present invention is not limited to the individual source used to amplify activated NK cells.
  • the HANK cell is obtained by in vitro activation of at least one of NK cells derived from the body: the patient's own NK cells, the patient's semi-matched NK cells, and an allogeneic NK cell.
  • the so-called patient's haplotype NK cells refer to NK cells from the relatives of the patient.
  • peripheral blood NK cells or the umbilical cord blood NK cells of the patient's relatives are not subject to the treatment limitation, as long as the blood transfusion infectious blood screening test is acceptable.
  • the so-called cell vesicles can be derived from natural cells or from engineered cells.
  • Engineering cells refer to the modification or recombination of the genetic material of the host cell by genetic engineering technology or cell fusion technology to obtain cells with stable inheritance and unique traits.
  • the cytokine carried by the cell vacant shell comprises at least one of IL-4, IL-7, IL-15, IL-21, CD19, CD64, CD86 and 4-1BBL.
  • the cell vacant shell carries IL-15, 4-1BBL and IL-21 cytokines.
  • it can be used to efficiently activate NK cells derived from the body in vitro to obtain HANK cells.
  • preparing the cell empty shell comprises: washing the cells to obtain washed cells; passing the washed cells through a hypotonic treatment or other methods commonly used in the industry to obtain the The cell is empty.
  • the inventors have found that the cell empty shell can be obtained quickly and efficiently by using the method, and the method is simple in operation, easy to control, and easy to realize mass production.
  • the cytokine carried by the cell vacant shell may be a cytokine expressed on the surface of the cell itself, or may be expressed in a cell by genetic engineering methods, for example, by transient transfection or stable expression.
  • the surface may also be a cytokine that adsorbs or crosslinks on the cell surface.
  • the cell for preparing the cell vesicle may be a primary cell such as PBMC or a passage cell such as K562 cell or the like.
  • the washing treatment comprises: suspending the cells in an isotonic solution to obtain a cell suspension; and centrifuging the cell suspension to obtain the washed cells.
  • the isotonic solution is pre-cooled to 4 degrees Celsius before suspending the cells in an isotonic solution.
  • the isotonic solution is an isotonic phosphate buffer (PBS) having a pH of 7.4.
  • PBS isotonic phosphate buffer
  • the hypotonic treatment comprises: suspending the washed cells in a hypotonic solution according to a predetermined volume ratio, and allowing the obtained cell suspension to stand for 2 hours to obtain a cell hypotonic treatment.
  • the cell hypotonic treatment is subjected to centrifugation to obtain the cell empty shell.
  • the predetermined volume ratio is 1:40
  • the hypotonic solution is a hypotonic Tris hydrochloric acid buffer.
  • the hypotonic solution is pre-cooled to 4 degrees Celsius before suspending the washed cells in a hypotonic solution.
  • obtaining the HANK cell comprises: isolating a monocyte from a peripheral blood, the PBMC comprising the in vivo NK cell; culturing the PBMC with a medium to which the cell vacant is added To Amplification is performed to activate NK cells in the PBMC to obtain the HANK cells, and the cell empty shell is obtained by the method for preparing a cell empty shell in any of the above embodiments.
  • activation of NK cells derived from the body by a large amount of expansion in vitro by the technique, obtaining HANK, and then returning to the patient for HANK cell therapy have been proven to have a good effect on various cancers.
  • the NK cells comprise at least 50% of said HANK cells.
  • the NK cells comprise at least 90% of the HANK cells.
  • the PBMC is cultured in a medium supplemented with a cell empty shell to amplify the in vivo NK cells in the activated PBMC to obtain HANK cells, including:
  • the PBMCs were cultured in an X-Vivo15 serum-free medium containing 200 IU/ml IL-2, the cell empty shell and 5% autologous plasma for 12-20 days, the number of empty cells added to the monocytes The ratio of the number is 1:1.
  • the X-Vivo15 serum-free medium and the cell vesicles are added at least once on days 4-8 of the culture.
  • the step of administering the NK cells in combination with the antibody drug is: administering the NK cells and the antibody drug sequentially or simultaneously.
  • the present invention provides a pharmaceutical composition comprising an antibody drug and NK cells, the NK cells comprising HANK cells, the obtaining of the HANK cells comprising: utilizing a cytokine Expansion of the NK cells from the body is activated by expansion outside the cell empty envelope.
  • the HANK cells and antibody drugs in this combination are mutually synergistic, and in vitro cell assays and in vivo mouse experiments indicate that treatment of the disease with the pharmaceutical composition can achieve significant therapeutic effects.
  • the diseases mentioned include cancer, viral infections and immune diseases.
  • the so-called patient has at least one of a cancer, a viral infection, and an immune disease.
  • the so-called patient is a cancer patient and the so-called antibody drug is a monoclonal antibody-targeted drug.
  • Monoclonal targeting drugs confer specificity to HANK cells.
  • rituximab allows HANK cells to kill leukemia cells.
  • Herceptin monoclonal antibody allows HANK cells to kill breast cancer cells; HANK cells are the monoclonal antibodies.
  • the drug provides enough active factors to effectively kill cancer cells, such as a large number of granzymes, perforin and the like.
  • HANK and monoclonal antibodies can increase the therapeutic effect of each other to achieve better treatment of cancer.
  • the antibody drug is a monoclonal antibody or a polyclonal antibody.
  • the two antigen binding sites on one antibody molecule may be the same or different, ie, bispecific antibodies, located at the ends of the arms, called antigen-binding fragments (Fab).
  • Fab antigen-binding fragments
  • the so-called antibody drug is Fc-including The complete monoclonal antibody of the segment.
  • the ratio of HANK cells to antibody drugs in NK cells in the pharmaceutical composition is 2 ⁇ 10 5 to 5 ⁇ 10 5 : 1 (pieces / ⁇ g).
  • the doctor or the drug instruction will give the amount according to the patient's condition.
  • In vitro tests and animal tests show that the specific ratio of HANK cells administered at the time of administration can significantly enhance the effect of treating cancer.
  • the present invention does not limit the source of the individual used to amplify the activated NK cells, as long as the transfusion infectious disease screening is acceptable.
  • the HANK cell is obtained by in vitro activation of at least one of NK cells derived from the body: NK cells of the patient, semi-matched NK cells of the patient, and NK cells which are not related to allogeneic .
  • the so-called patient's haplotype NK cells refer to NK cells from the relatives of the patient.
  • the patient's own PBMC is collected to prepare NK cells, preferably before conventional treatment; or the patient's relatives, that is, the semi-matched PBMCs are prepared to prepare NK cells; or the unrelated individual PBMCs are used to prepare NK cells or cord blood to prepare NK cells, It is not subject to this restriction.
  • the cytokine carried by the cell vacant shell comprises at least one of IL-4, IL-7, IL-15, IL-21, CD19, CD64, CD86 and 4-1BBL.
  • the cell vacant shell carries IL-15, 4-1BBL and IL-21 cytokines. In this way, NK cells in vivo can be efficiently expanded in vitro to obtain HANK cells.
  • a method of enhancing an ADCC effect in a patient being treated with an antibody drug comprising the step of administering a combination of NK cells and an antibody drug, the NK cells comprising HANK cells,
  • the acquisition of HANK cells involves the activation of NK cells in vivo using an empty envelope of cells with cytokines.
  • the so-called patient has at least one of a cancer, a viral infection, and an immune disease.
  • the so-called patient is a cancer patient and the so-called antibody drug is a monoclonal antibody-targeted drug.
  • Monoclonal targeting drugs confer specificity to HANK cells.
  • rituximab allows HANK cells to kill leukemia cells.
  • Herceptin monoclonal antibody allows HANK cells to kill breast cancer cells; HANK cells are the monoclonal antibodies.
  • the drug provides enough active factors to effectively kill cancer cells, such as a large number of granzymes, perforin and the like.
  • HANK cells and monoclonal antibodies can increase the therapeutic effect of each other to better treat cancer.
  • the antibody drug is a monoclonal antibody or a polyclonal antibody.
  • the two antigen binding sites on one antibody molecule may be the same or different, ie, bispecific antibodies, located at the ends of the arms, called antigen-binding fragments (Fab).
  • Fab antigen-binding fragments
  • the so-called antibody drug is an intact monoclonal antibody comprising an Fc segment.
  • the ratio of HANK cells to antibody drugs in the NK cells administered in combination is 2 ⁇ 10 5 to 5 ⁇ 10 5 : 1 (pieces / ⁇ g).
  • the doctor or the drug manual will give an amount according to the patient's condition. In vitro tests and animal tests show that the combined administration of this ratio of HANK cells at the time of administration can significantly enhance the effect of treating cancer.
  • the present invention does not limit the PBMC-derived individuals used to amplify activated NK cells in vivo, as long as the transfusion infectious disease is screened.
  • the HANK cells are obtained by at least activating one of the following NK cells in vitro: NK cells of the patient, semi-matched NK cells of the patient, and NK cells that are not allogeneic.
  • the so-called patient's haplotype NK cells refer to NK cells from the relatives of the patient.
  • NK cells in the patient's own PBMC are collected, preferably prior to conventional treatment; NK cells in the patient's relatives, ie, semi-matched PBMCs, or NK cells or cord blood NK cells in PBMCs of unrelated individuals are collected. It is not subject to this restriction.
  • the cytokine carried by the cell vacant shell comprises at least one of IL-4, IL-7, IL-15, IL-21, CD19, CD64, CD86 and 4-1BBL.
  • the cell vacant shell carries IL-15, 4-1BBL and IL-21 cytokines. In this way, it is possible to efficiently activate and activate NK cells in vivo in vitro to obtain HANK cells.
  • HANK cells with antibody drugs, particularly monoclonal antibody targeting agents, to increase the lethality of monoclonal antibodies, and to enable a broad spectrum of non-specific HANK cells to target tumor cells and viruses.
  • antibody drugs particularly monoclonal antibody targeting agents
  • the function of infecting cells, the two can synergize each other.
  • Figure 1 shows the ADCC effect of HANK cells in combination with rituximab on SUDHL-4 lymphoma cells in one embodiment of the invention.
  • Figure 2 shows the efficacy of HANK cells in combination with rituximab in one embodiment of the invention on mouse lymphoma.
  • Figure 3 shows the ADCC effect of HANK cells in combination with Herceptin monoclonal antibody on MDA-MB-435 breast cancer cells in one embodiment of the invention.
  • Figure 4 shows the efficacy of HANK cells in combination with Herceptin monoclonal antibody in mouse breast cancer in one embodiment of the invention.
  • Figure 5 shows the ADCC effect of HANK cells in combination with GPC3 mAb on HepG2 liver cancer cells in one embodiment of the invention.
  • Figure 6 shows the effect of HANK cells in combination with GPC-3 monoclonal antibody on liver cancer in mice in one embodiment of the present invention.
  • Figure 7 shows the ADCC effect of HANK cells in combination with mC3 monoclonal antibody on JEV-infected BHK cells in one embodiment of the invention.
  • Figure 8 shows the protective effect of HANK cells in combination with mC3 monoclonal antibody on JEV-infected mice in one embodiment of the invention.
  • RPMI 8866 which is a human B lymphoblastoid cell line according to the following procedure. details as follows:
  • RPMI8866 cells were suspended in 3-fold amount of isotonic PBS pre-cooled to 4 ° C and pH 7.4, centrifuged at 1500 rpm for 10 minutes at 4 ° C, the supernatant was removed, and washing was repeated 1-3 times to obtain washed RPMI 8866.
  • RPMI 8866 cells were added to a hypotonic Tris-HCl buffer pre-cooled to 4 ° C and a concentration of 10 mmol/L at a ratio of 1:40, while slowly stirring, then, The mixture was allowed to stand in a refrigerator at 4 ° C for 2 hours to completely lyse the cells; then, the cells were centrifuged at 9000 rpm for 10 minutes at 4 ° C to precipitate the cells, and the washing was further repeated and centrifuged 3-5 times.
  • RPMI8866 cell empty shell namely RPMI 8866-empty shell. Then, it is divided into 2 ⁇ 10 7 cell empty shells/ml, and is frozen in a refrigerator at minus 80 degrees Celsius; or after being freeze-dried, it is stored frozen in a refrigerator at 4 degrees Celsius.
  • PBMC can be cultured in X-Vivo15 serum-free medium supplemented with IL-2 and RPMI 8866-shell and 5% autologous plasma, which can activate and activate NK cells in large quantities;
  • the number of NK cells can be expanded by hundreds to thousands of times, and the purity of NK cells can be increased from 10% in PBMC to above 80-90%;
  • the activated NK cells expanded in vitro are HANK cells, which can be used freshly or frozen in a -80 degree refrigerator or liquid nitrogen for use as effector cells in subsequent in vitro killing assays, in animal experiments alone or Combined with monoclonal antibody, it plays a role in anti-cancer and anti-virus.
  • lymphocyte complete medium containing about 200 IU/ml IL-2 and autologous plasma 1-10% was added to the isolated PBMC to prepare a cell suspension (about 5 ⁇ 10 6 cells). Lymphocytes), added to the T75 flask; simultaneously added 2 ⁇ 10 7 RPMI 8866 - empty shell, placed in a saturated humidity, 37 ° C, 5.0% CO 2 incubator;
  • each culture bag contains about 640 ml of the complete lymphocyte culture solution
  • NK cells in PBMC can be cultured at different times depending on the number of cells required, for example, continuous culture for 18 days or 20 days.
  • NK cell culture medium NKEM, X-Vivo15 containing IL-2 and 5% autologous plasma
  • test set the maximum release hole, the negative control hole, the natural killing resistance, and the ADCC hole;
  • a) Establishment of a tumor model of human diffuse large B-cell lymphoma subcutaneous inoculation of 10 ⁇ 7 cells in SCID mice can successfully establish a diffuse large B-cell lymphoma (DLBCL) xenograft model, and the tumor formation rate 70%, the histological performance of the tumor is similar to human DLBCL.
  • DLBCL diffuse large B-cell lymphoma
  • SUDHL-4 is a human GCB-like DLBCL cell line.
  • the cells with an initial density of 2.5 ⁇ 10 ⁇ 5/ml were placed in a T25 cell culture flask containing RPMI1640 medium containing 10% FBS, 100 U/ml penicillin, 100 ⁇ g/ml glutamine, and 30 ⁇ g/ml glutamine.
  • mice Female, 5 weeks old, weighing 16-20g, were randomly divided into groups. The feeding and experiment were carried out in a SPF class rat room with constant temperature (20-26 °C) and constant humidity (50%-56%). . The mice were placed in a laminar flow box with a cover mouse box, the air was filtered by medium efficiency, and the standard pellet feed was fed, and all the items in contact with the rats were previously sterilized;
  • mice (10 per group) cell suspension containing 107 cells were inoculated subcutaneously on one side of the rib 0.1ml; normal control group (10 per group) were injected subcutaneously in the right side of the rib 0.1ml PBS.
  • mice During the experiment, the general condition, tumor formation and tumor growth of the mice were observed daily. Body weight and tumor length and diameter were measured daily, and tumor volume was calculated (calculation method: ⁇ /6 ⁇ length ⁇ width ⁇ height); when the tumor reached 1200 mm 3 , it was regarded as the human end point. After the mice were anesthetized and the neck was sacrificed, the tumors were observed at various parts of the body surface; then the animals were dissected and the internal organs and lymph node metastasis were observed.
  • Human breast cancer MDA-MB-435 cell line nude mouse xenograft model fully humidified with 37 U/ml gentamicin, 10% inactivated calf serum MEM medium at 37 ° C and 5% CO 2
  • the human breast cancer cell line MDA-MB-435 was used for further use.
  • MDA-MB-435 human breast cancer cells were inoculated into the fat pad of the second nipple on the left side of nude mice, and the inoculum amount was 0.1 ml/cell (the number of cells was 1 ⁇ 10 6 /piece). After 2 weeks, the subcutaneous lesions were clearly randomized when they reached the growth of the mass. The administration time was 10 weeks, and the animals were sacrificed by cervical dislocation 3 days after the withdrawal.
  • JEV SA14 is administered through the brain of the suckling rat for 3 consecutive generations of poisoning
  • mice 8 g of Balb/c mice of 3 weeks old were used, and the brain suspension of JEV-infected rats was inoculated intraperitoneally, and the measurement was 10 ⁇ 5 LD50;
  • Rituxoxib is an anti-CD20 mAb that binds to the Fc receptor CD16 on NK cells and kills CD20 positive cells, and has been used in clinical treatment of leukemia for many years in many countries.
  • the results are shown in Figure 1.
  • the killing rate of HANK cells on K562 cells is about 92%, and the killing rate on SUDHL-4 cells is about 52%.
  • Rituximab does not affect the killing effect of HANK cells on K562 cells, but The killing effect of HANK cells on SUDHL-4 cells was greatly increased, from 52% to 87%.
  • the tumor volume of the saline control group was 1129 mm ⁇ 3
  • the tumor volume of the HANK cell treatment group was 224 mm ⁇ 3
  • the rituximab-targeted treatment group was 315 mm ⁇ 3, HANK cells + rituximab.
  • Herceptin is an anti-EGFR Her2 monoclonal antibody that binds to Her2 positive cells and kills EGFR-positive cells and has been used in clinical treatment of breast cancer for many years in many countries.
  • the results are shown in Figure 3.
  • the killing rate of HANK cells to K562 cells is about 92%, and the killing rate of MDA-MB-435 cells is about 63%.
  • Herceptin monoclonal antibody does not affect the killing effect of HANK cells on K562 cells.
  • the killing effect of HANK cells on MDA-MB-435 cells was greatly increased, from 63% to about 91%.
  • the results are shown in Figure 4: 14 days after treatment, the saline control group had a tumor volume of 724 mm ⁇ 3, the HANK cell treatment group had a tumor volume of 121 mm ⁇ 3, and the Herceptin monoclonal antibody targeted treatment group was 312 mm ⁇ 3, HANK cells + Herceptin combination. Treatment group 51mm ⁇ 3. The smaller the tumor volume after treatment, the better the efficacy.
  • 1G12 is an anti-GPC3 monoclonal antibody that inhibits the proliferation of GPC3-positive hepatoma cells in vitro and has a good therapeutic effect on transplanted hepatocellular carcinoma (HCC) in nude mice.
  • HCC hepatocellular carcinoma
  • the killing rate of HANK cells to K562 cells is about 92%, for SUDHL-4 cells.
  • the killing rate was about 52%; 1G12 monoclonal antibody did not affect the killing effect of HANK cells on K562 cells, but greatly increased the killing effect of HANK cells on HepG2 cells, from 48% to 85%.
  • the tumor volume of the saline control group was 936 mm ⁇ 3
  • the tumor volume of the HANK cell treatment group was 185 mm ⁇ 3
  • the target treatment group of 1G12 monoclonal antibody was 423 mm ⁇ 3
  • the HANK cell +1G12 combination treatment group was 77 mm ⁇ . 3.
  • mC3 is an anti-JEV (Japanese encephalitis virus) monoclonal antibody, which has the activity of neutralizing JEV alone and has a protective effect on mice infected with JEV.
  • JEV Japanese encephalitis virus
  • HANK cells were used as effector cells, K562 cells and JEV-infected BHK cells were used as target cells, and the natural killing effect of HANK cells on two kinds of target cells and the anti-JEV mC3-mediated ADCC effect were determined.
  • the results are shown in Figure 7.
  • the killing rate of HANK cells to K562 cells was about 92%, and the killing rate of JEV-BHK cells was about 62%.
  • the mC3 monoclonal antibody did not affect the killing effect of HANK cells on K562 cells, but greatly improved.
  • the killing effect of HANK cells on JEV-BHK cells increased from 62% to about 88%.
  • mice On the 5th day after JEV infection, some mice were nearly dead. The infected mice were divided into 4 groups of 10 animals each. One group was treated with HANK cells alone, and each mouse was intravenously infused with 1X10 ⁇ 7HANK cells once a week. The other group was treated with mC3 monoclonal antibody alone. Each mouse was intravenously infused with 40 ⁇ g once a week; The group was treated with HANK cells combined with mC3 monoclonal antibody, intravenously infused with 1X10 ⁇ 7HANK cells plus 40 ⁇ g mC3 monoclonal antibody once a week; group 4 was a saline control group.

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Abstract

L'invention concerne un procédé permettant d'améliorer la capacité de tuer une cellule anormale, une composition pharmaceutique et un procédé permettant d'améliorer l'ADCC chez un patient subissant une thérapie médicamenteuse par anticorps. Le procédé de traitement d'un patient comprend une étape d'administration au patient d'une cellule NK en combinaison avec un médicament anticorps. La cellule NK comprend une cellule HANK. L'acquisition de la cellule HANK comprend : l'utilisation d'une enveloppe cellulaire vide portant une cytokine pour effectuer une activation in vitro d'une cellule NK in vivo. La cellule NK comprenant la cellule HANK est administrée au patient en combinaison avec le médicament anticorps. La cellule HANK et le médicament anticorps permettent d'améliorer mutuellement leurs capacités à tuer les cellules anormales, améliorant ainsi significativement l'effet du traitement.
PCT/CN2015/094143 2015-11-09 2015-11-09 Procédé permettant d'améliorer la capacité de tuer des cellules anormales et composition pharmaceutique WO2017079881A1 (fr)

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US10428305B2 (en) 2014-05-15 2019-10-01 National University Of Singapore Modified natural killer cells that express IL15 and uses thereof
US11141436B2 (en) 2019-03-05 2021-10-12 Nkarta, Inc. Immune cells engineered to express CD19-directed chimeric antigen receptors and uses thereof in immunotherapy
US11365236B2 (en) 2017-03-27 2022-06-21 Nkarta, Inc. Truncated NKG2D chimeric receptors and uses thereof in natural killer cell immunotherapy

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