CN107126563A - Composition of antibody of the signal path of blocking VEGF containing low dosage and application thereof - Google Patents

Composition of antibody of the signal path of blocking VEGF containing low dosage and application thereof Download PDF

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Publication number
CN107126563A
CN107126563A CN201610107589.4A CN201610107589A CN107126563A CN 107126563 A CN107126563 A CN 107126563A CN 201610107589 A CN201610107589 A CN 201610107589A CN 107126563 A CN107126563 A CN 107126563A
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pharmaceutical composition
signal path
active component
tumor
vegf signal
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CN107126563B (en
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杨林
黄玉辉
游凤涛
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PERSONGEN BIOMEDICINE (SUZHOU) CO Ltd
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PERSONGEN BIOMEDICINE (SUZHOU) CO Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • 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
    • 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
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
    • 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
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
    • 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
    • A61K39/46—Cellular immunotherapy
    • A61K39/463—Cellular immunotherapy characterised by recombinant expression
    • A61K39/4631—Chimeric Antigen Receptors [CAR]
    • 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
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A61K39/464402—Receptors, cell surface antigens or cell surface determinants
    • A61K39/464411—Immunoglobulin superfamily
    • A61K39/464412—CD19 or B4
    • 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
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A61K39/464452—Transcription factors, e.g. SOX or c-MYC
    • A61K39/464453—Wilms tumor 1 [WT1]
    • 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
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A61K39/464469—Tumor associated carbohydrates
    • A61K39/46447—Mucins, e.g. MUC-1
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/31—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterized by the route of administration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
    • A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the cancer treated
    • A61K2239/48—Blood cells, e.g. leukemia or lymphoma
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment

Abstract

The invention provides a kind of composition of the signal paths of VEGF containing low dosage blocking agent and application thereof.Specifically, the invention provides the VEGF signal paths blocking agent of low dosage for preparing the purposes in immunotherapy of tumors accelerator, and the composition containing low dosage VEGF signal path blocking agents.The experiment of the present invention shows that the VEGF signal path blocking agents of low dosage are capable of the normalization of induced tumor blood vessel, significantly improve the effect of immunotherapy of tumors, and can promote infiltration of the T lymphocytes to tumor tissues.

Description

Composition of antibody of the signal path of blocking VEGF containing low dosage and application thereof
Technical field
The present invention relates to field of medicaments, relate more specifically to one kind signal path blocking agents of VEGF containing low dosage, Composition of antibody such as blocking VEGF signal path and application thereof.
Background technology
Folkman is reported first within 1971, and the volume of entity tumor is more than 2-3mm3When, simple oxygen expands Dissipating just can not support the growth of knurl body, it is necessary to have new vessels formation to support its continued growth.This A little tumor neogenetic blood vessels compared with normal blood vessels, have blood vessel dilatation, tortuous, cystic structures formation phenomenon, Produce the branch of random connection, vessel density skewness；Caliber regulation mechanism, which is lost, causes abnormal point Stream, vasopermeability and vascular space are dramatically increased.Abnormal tumor neogenetic blood vessels eventually result in blood vessel Skewness, subregion blood flow cessation aggravates the formation of tumor hypoxia microenvironment, then influences tumour Gene phenotype, the activation new vessels generation factor, including VEGF (VEGF), promote tumour The generation of new vessels, and cause the unstability of tumor cell gene, activate the existence of some tumour cells because Son, causes tolerance of the tumour cell to radiation and chemotherapy, further promotes metastases.Importantly, Tumor vessel lacks high endothelials venules (HEV), therefore to invade profit not enough for T cell so that Immunotherapy Strategy without Method is implemented effectively.
The T lymphocytes of Chimeric antigen receptor (chimeric antigen receptor, CAR) genetic modification (hereinafter referred to as CAR-T cells) treatment malignant tumour technology is elected as first of the achievement of scientific invention in 2013, The complete remission rate more than 90% is reached in recurrence, refractory acute lymphatic leukemia treatment, now The technology of this representative world forefront and latest development trend is generalized to including entity tumor Various treatments of cancer experiment in.However, due to the tumor vessel and the tumour of inhibitive ability of immunity of dysfunction The presence of microenvironment, the CAR-T cells for seriously hindering venous re-transfusion are gone back to the nest to tumor focus position and infringement Existence of the CAR-T cells in inside tumor so that this revolutionary technology is on treatment of solid tumor Effect is still very undesirable so far, and CAR-T cells are applied to the treatment of entity tumor still face in the world Face serious challenge.Therefore, this at present field in the urgent need to exploitation it is new, effective controlled using CAR-T cells The method for treating entity tumor.
The content of the invention
It is an object of the invention to provide a kind of CAR-T cells based on tumor vessel normalization and intervention strategies Treat the pharmaceutical composition of entity tumor.
In the first aspect of the present invention, there is provided a kind of pharmaceutical composition for being used to treat entity tumor, the medicine Compositions include the first active component and the second active component, wherein, the first active component is VEGF signal paths Blocking agent, the second active component is immunotherapy of tumors agent,
And described pharmaceutical composition is unit dosage form, and the first active component contains described in each unit dosage form 0.1 to 0.5 (the preferably 0.15-0.4, or 0.2-0.25) for routine dose is measured, wherein, the conventional agent Measure as 200-400mg/50kg body weight.
In another preference, the routine dose is the total conventional agent for the first active component being administered every time Amount, such as 200-400mg/50kg body weight.
In another preference, the content of the first active component corresponds to each in a described unit dosage form The 0.1 to 0.5 of total routine dose of first active component of administration.
In another preference, the total content of the first active component corresponds to every in n described unit dosage form The 0.1 to 0.5 of total routine dose of first active component of secondary administration, wherein n is 2,3,4 or 5.
In another preference, n described unit dosage form is identical or different.
In another preference, the content of the first active component is described in described unit dosage form 10-160mg/ agent, preferably 10-100mg/ agent, more preferably 10-80mg/ agent, such as 10,20,30,40, 50th, 60,70,80mg/ agent.
In another preference, described unit dosage form dosage is 20-160mg/50kg body weight, preferably Ground is 30-100mg/50kg body weight, by first active ingredient.
In another preference, described unit dosage form is the formulation of the active component comprising dosage unit.
In another preference, described dosage unit refers to the single administration dosage of active component.
In another preference, the first described active component is antibody or micromolecular compound.
In another preference, described VEGF signal paths blocking agent include blocking VEGF signal path antibody and Tyrosine kinase inhibitor (TKI).
In another preference, the antibody of described blocking VEGF signal path includes：Thunder not Lu Dankang (DC101 / ramucirumab), bevacizumab (Avastin) or its combination.
In another preference, described tyrosine kinase inhibitor includes：Sutent (Sunitinib), Nexavar (Sorafinib) or its combination.
In another preference, the dosage of the antibody of the blocking VEGF signal path is 0.1-4mg/kg, Preferably 0.2-2.5mg/kg, more preferably 0.5-1mg/kg.
In another preference, the immunotherapy of tumors agent includes CAR-T cells, CAR-NK cells, tumour Vaccine and immunologic test point inhibitor.
In another preference, the dosage of the CAR-T cells is 1x107-5x108CAR-T cells/kg.
In another preference, first active component and the second active component are mixing or individualism.
In another preference, the first described active component and the total content of the second active component be 70~ 100wt%, preferably 80~100wt%, are more preferably 90~100wt%, by described pharmaceutical composition The gross weight meter of active component.
In another preference, described entity tumor is selected from the group：Tumor of breast, liver tumour, lung neoplasm, essence Capsule adenoncus knurl, cancer of pancreas, stomach cancer, colorectal cancer, the cancer of the esophagus, glioma or its combination.
In another preference, described immunotherapy of tumors include CAR-T cell therapies, CAR-NK cell therapies, Tumor vaccine therapy, immunity inspection point inhibitor for treating or its combination.
In another preference, described immunity inspection point inhibitor includes PD-1 antibody and CTLA-4 antibody.
In another preference, described pharmaceutical composition also includes pharmaceutically acceptable carrier.
In another preference, the formulation of described pharmaceutical composition includes injection and oral agents.
In the second aspect of the present invention there is provided a kind of purposes of the VEGF signal path blocking agents of low dosage, use In preparing a pharmaceutical composition or medicine box, described pharmaceutical composition or medicine box are used for the treatment for improving immunotherapy of tumors Effect,
Wherein, low dosage refers to the dosage A1 and conventional administration dosage A0 of the VEGF signal paths blocking agent Compare, A1≤1/2A0, preferably A1≤1/4A0.
In another preference, described VEGF signal paths blocking agent includes the antibody of blocking VEGF signal path.
In another preference, the dosage of the antibody of the blocking VEGF signal path is 0.1-4mg/kg, Preferably 0.2-2.5mg/kg, more preferably 0.5-1mg/kg.
In another preference, described medicine box includes the VEGF in 2-4 unit dosage form, each unit formulation Signal path blocking agent, such as content of the antibody of blocking VEGF signal path for routine dose 0.1 to 0.5 (compared with Good ground 0.15-0.4, or 0.2-0.25), wherein, the routine dose is 200-400mg/50kg body weight.
In another preference, described pharmaceutical composition is the VEGF signals in unit dosage form, the unit dosage form Path blocking agent, such as content of the antibody of blocking VEGF signal path for routine dose 0.1 to 0.5 (preferably 0.15-0.4, or 0.2-0.25), wherein, the routine dose is 200-400mg/50kg body weight.
In the third aspect of the present invention, there is provided a kind of medicine box for being used to improve immunotherapy of tumors effect, institute Stating medicine box includes：
(a) the first pharmaceutical composition, first pharmaceutical composition includes VEGF signal path blocking agents, and pharmacy Upper acceptable carrier, and first pharmaceutical composition is unit dosage form, and the medicine box includes 2-4 Individual unit dosage form, wherein the content of the VEGF signal path blocking agents in the unit dosage form is the 0.1 of routine dose To 0.5 (preferably 0.15-0.4, or 0.2-0.25), wherein, the routine dose is 200-400mg/50kg Body weight；
(b) the second optional pharmaceutical composition, the second described pharmaceutical composition is immunotherapy of tumors agent；
(c) specification.
In another preference, the second described pharmaceutical composition is cell preparation.
In another preference, the second described pharmaceutical composition is CAR-T preparations.
In another preference, described CAR-T preparations include autologous or allosome CAR-T preparations.
In another preference, described the first pharmaceutical composition and the second pharmaceutical composition is each independent.
In another preference, the first described pharmaceutical composition and the second pharmaceutical composition are respectively positioned at different In packaging or container.
In another preference, the treatment method described in fourth aspect present invention has been recorded in the specification.
In the fourth aspect of the present invention, there is provided a kind of method for the effect for improving immunotherapy of tumors, including step Suddenly：
(I) the VEGF signal path blocking agents of low dosage are applied to the object needed；
(II) immunotherapy of tumors is carried out to the object,
Wherein, the low dosage refers to dosage A1 and the conventional administration agent of the VEGF signal paths blocking agent Amount A0 is compared, A1≤1/2A0, preferably A1≤1/4A0.
In another preference, described VEGF signal paths blocking agent includes the antibody of blocking VEGF signal path.
In another preference, in step (I), including twice or thrice to the object using blocking VEGF letter The antibody of number path, the interval time between double administration is t1.
In another preference, described t1 is 3-21 days, and preferably 4-15, is more preferably 7-14 days.
In another preference, after last is applied in step (I), certain interval of time t2 carries out the step (II)。
In another preference, described t2 is 3-21 days, and preferably 4-15, is more preferably 7-14 days.
In another preference, described object includes people and non-human mammal, and (such as rodent and spirit are long dynamic Thing).
In another preference, step is also included after step (II)：
(III) antibody of the blocking VEGF signal path of low dosage is applied to the object.
In another preference, after last is applied in step (II), certain interval of time t3 carries out the step (III)。
In another preference, described t3 is 3-21 days, and preferably 4-15, is more preferably 7-14 days.
It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the invention and (such as implementation below Example) in specifically describe each technical characteristic between can be combined with each other, so as to constitute new or preferred skill Art scheme.As space is limited, no longer tire out one by one herein and state.
Brief description of the drawings
Fig. 1 shows that the anti-angiogenic medicaments of low dosage being capable of induced tumor blood vessel normalization.Wherein, scheme 1A shows representational immunohistochemical staining figure, and endothelial cell marker thing CD31 molecules show red fluorescence, Perithelial cells label NG2 molecules show green fluorescence；Figure 1B shows the statistical result of microvessel density, Fig. 1 C show the statistical result of pericyte coverage rate.
Fig. 2 shows that the anti-angiogenic medicaments treatment of low dosage can strengthen the cancer suppressing action of tumor vaccine. In figure, D13, D16, D19, D22 are represented the 13rd after being inoculated with first respectively, 16,19,22 days, D10 tables Show injection 10mg/kg anti-angiogenic medicaments DC101.
Fig. 3 shows that the anti-angiogenic medicaments treatment of low dosage can promote the tumor-infiltrated of T cell.Its In, Fig. 3 A show the tumor-infiltrated situation of CD4+T cells；Fig. 3 B show the tumour leaching of CD8+T cells Profit situation.
Fig. 4 shows that the anti-angiogenic medicaments of routine dose treat the influence to tumor vaccine therapy.In figure, D7, D10, D13, D16 are represented the 7th after being inoculated with first respectively, 10,13,16 days, D40 represents to inject 40mg/kg Anti-angiogenic medicaments DC101.
Fig. 5 shows the tumor patient receiving by low dosage blocking VEGF signal path medical preconditioning The pathological section analysis result of CAR-T cell PCIs.
Embodiment
The present inventor is by in-depth study extensively, first it was unexpectedly observed that the VEGF signals of low dosage lead to Road blocking agent, such as antibody of blocking VEGF signal path can significantly improve the effect of immunotherapy of tumors, promote Enter infiltration of the T lymphocytes to tumor tissues.Experiment shows, is arrived CAR-T cell infusions using intervention strategies Inside solid tumor tissue, while coordinating the antibody of the blocking VEGF signal path of low dosage, induced tumor blood There is obvious necrosis in the normalization of pipe, injection site tumour.On this basis, the present invention is completed.
VEGF
VEGF (English：Vascular endothelial growth factor, referred to as： VEGF), early stage is also referred to as vascular permeability factor (English：Vascular permeability factor, letter Claim：VPF), it is the specific HBGF (heparin-binding of vascular endothelial cell Growth factor), can in vivo induction of vascular it is newborn (induce angiogenesis in vivo).
Research shows that the volume of entity tumor is more than 2-3mm3When, simple oxygen diffusion just can not have been propped up Support the growth of knurl body, it is necessary to there is new vessels formation to support its continued growth.These tumor neogenetic blood vessels Compared with normal blood vessels, there are blood vessel dilatation, tortuous, cystic structures formation phenomenon, produce random connection Branch, vessel density skewness；Caliber regulation mechanism, which is lost, causes abnormal shunting, vasopermeability Dramatically increased with vascular space.It is uneven that abnormal tumor neogenetic blood vessels eventually result in vascular distribution, portion Subregion blood flow cessation, aggravates the formation of tumor hypoxia microenvironment, then influences oncogene phenotype, activation New vessels generates the factor, including VEGF (VEGF), promotes tumor neovasculature generation, And the unstability of tumor cell gene is caused, some tumour cell Survival Factors are activated, cause tumour cell Tolerance to radiation and chemotherapy, further promotes metastases.
VEGF signal path blocking agents
As used herein, " VEGF signal paths blocking agent " refers to suppress by the signal path of blocking VEGF VEGF blood vessel adjusting function.VEGF signal paths blocking agent includes the antibody and junket ammonia of blocking VEGF signal path Acid kinase inhibitor (TKI).
" antibody of blocking VEGF signal path " refer to specifically bind VEGF acceptor or its Part, is a kind of typical anti-angiogenic medicaments.
The antibody drug of blocking VEGF signal path clinically has 2 kinds at present：Thunder not Lu Dankang And bevacizumab (Avastin) (Ramucirumab).
Lu Dankang (Ramucirumab) is not a kind of complete human monoclonal antibody to thunder, and its corresponding mouse source is single Clonal antibody is DC101.It specifically can be combined with VEGF R2 (VEGFR2/KDR), So as to prevent acceptor from being activated, so it is a kind of VEGFR2 inhibitor.Thunder not Lu Dankang by U.S.'s food medicine Product Surveillance Authority (FDA) ratifies to be used to treat some solid tumors, including stomach cancer and Metastatic Nsclc (NSCLC)。
Bevacizumab (Avastin) is a kind of Humanized monoclonal IgG1 antibody of restructuring, by combining VEGF parts To block the acceptor (Flt-1 and KDR) of itself and endothelial cell surface to combine, so as to suppress VEGF Biological activity and work.Bevacizumab is ratified to be used to treat a series of by food and medicine Surveillance Authority of the U.S. Cancer, including colon cancer, lung cancer, kidney and the cancer of the brain etc..
Clinically, Ramucirumab routine dose is 8mg/kg；Avastin routine dose is 5mg/kg. Mouse test results in embodiment, applicant thinks that 1/5 to 1/10 is preferably dosage, but be not precluded within The different phase of different tumours and tumour and its different agent are had when being combined from different immunotherapy of tumors means Measure scope.
Compound medicament composition and medicine box
The invention provides containing the first active component VEGF signal path blocking agents, such as blocking VEGF signal leads to The antibody on road, and the second active component immunotherapy of tumors agent；And the compound of pharmaceutically acceptable carrier Pharmaceutical composition.This kind of carrier includes (but being not limited to)：Salt solution, buffer solution, glucose, water, glycerine, Ethanol, pulvis, and combinations thereof.Pharmaceutical preparation should match with administering mode.The pharmaceutical composition of the present invention Injection form can be made into, such as aqueous solution with physiological saline or containing glucose and other assistant agents passes through It is prepared by conventional method.The pharmaceutical composition of such as tablet and capsule etc, can be carried out by conventional method Prepare.Pharmaceutical composition such as injection, solution, tablet and capsule are preferably aseptically manufactured.The present invention's Drug regimen, which can also be made into pulvis, is used for Neulized inhalation.A kind of preferred formulation is ejection preparation.In addition, Pharmaceutical composition of the present invention can be also used together with other therapeutic agents.
Present invention also offers a kind of medicine box for being used to improve immunotherapy of tumors effect, the medicine box includes：
(a) the first pharmaceutical composition, first pharmaceutical composition includes VEGF signal path blocking agents, preferably Include the antibody of blocking VEGF signal path, and pharmaceutically acceptable carrier, and first drug regimen Thing is unit dosage form, and the medicine box includes 2-4 unit dosage form, wherein the VEGF in the unit dosage form The content of signal path blocking agent is 0.1 to 0.5 (preferably 0.15-0.4, or 0.2-0.25) of routine dose, Wherein, the routine dose is 200-400mg/50kg body weight；
(b) the second optional pharmaceutical composition, the second described pharmaceutical composition is immunotherapy of tumors agent；
(c) specification.
The pharmaceutical composition or medicine box of the present invention is applied to treatment tumour, and be particularly suitable for use in treatment entity tumor.
It should be noted that the pharmaceutical composition or medicine box of the present invention contain the VEGF signal paths resistance of low dosage The antibody of disconnected agent, such as blocking VEGF signal path, wherein, low dosage refers to the blocking VEGF signal path The dosage A1 of antibody is compared with conventional administration dosage A0, A1≤1/2A0, preferably A1≤1/4A0.
In another preference, pharmaceutical composition of the invention is unit dosage form, described in each unit dosage form The content of active component for daily dose 0.1 to 1 (or 0.25-1, or 0.5-1), wherein first activity The daily dose of composition is 20-100mg (for 50kg people).
In another preference, the dosage of the antibody of the blocking VEGF signal path is 0.1-4mg/kg, Preferably 0.2-2.5mg/kg, more preferably 0.5-1mg/kg.
Certainly, the effective dose of active component used can with administration pattern and disease to be treated it is serious Degree etc. and be varied from.
Treatment method
Present invention also offers entity tumor is controlled with the two kinds of active components or corresponding medicine of the present invention The method for the treatment of, it is included to mammal using the first active component blocking VEGF signal path of low dosage Antibody and the second active component immunotherapy of tumors agent, or apply containing first active component and the The pharmaceutical composition of two active components.
, can be pharmaceutically acceptable with one or more when two kinds of active components of the present invention is used for such use Carrier or excipient mixing, such as solvent, diluent, and sterile injectable solution or suspension can be used Liquid form (containing about 0.05-5% suspending agents in isotonic medium) carries out parenteral routes.For example, these medicines Preparation can contain the about 0.01-99% mixed with carrier, more preferably about 0.1%-90% (weight) active component.
The two kinds of active components or pharmaceutical composition of the present invention can be administered by conventional route, wherein wrapping Include (but being not limited to)：Intramuscular, intraperitoneal, intravenous, subcutaneous, intracutaneous, oral, knurl are interior or part is given Medicine.It is preferred that method of administration include knurl in administration or intravenous administration.
In terms of the position for being easy to be administered, pharmaceutical composition preferably is fluid composition.
In addition, the two kinds of active components or medicine of the present invention can also with the medicine of other treatment cancer (such as cis-platinum, Taxol etc.) combination.
Main advantages of the present invention include：
(a) the VEGF signal path blocking agents of low dosage, such as antibody of blocking VEGF signal path can be notable Improve the effect of immunotherapy of tumors.
(b) the VEGF signal path blocking agents of low dosage, such as antibody of blocking VEGF signal path can promote T Infiltration of the lymphocyte to tumor tissues.
(c) the VEGF signal path blocking agents of low dosage, such as antibody of blocking VEGF signal path, may additionally facilitate The effect of other tumor therapeuticing methods, such as chemotherapy and radiation.
(d) drug regimen of the invention is applied to the treatment of entity tumor.
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate The present invention rather than limitation the scope of the present invention.The experiment side of unreceipted actual conditions in the following example Method, generally according to normal condition, or according to the condition proposed by manufacturer.Unless otherwise indicated, otherwise Percentage and number are calculated by weight.
Versatile material
In following embodiments, blocking VEGF signal path medicine DC101 is purchased from Bio X Cell companies, is used as The rabbit anti-mouse igg monoclonal antibody of control is purchased from Jackson ImmunoResearch companies.MCaP0008 tumour cells Vaccine is the MCaP0008 tumour cells of mitomycin processing, and the tumor vaccine is thin by internal antigen submission Born of the same parents induce the immune response that MCaP0008 tumour antigens are special.Anti-angiogenic medicaments bevacizumab is by hospital There is provided；CAR-T cells for cellular immunotherapy are produced by Bo Shengji companies.
Embodiment 1
The anti-angiogenic medicaments of low dosage are on tumor vascular influence
Original position MCaP0008 breast cancer models are built, when MCaP0008 breast cancer mouse diameter of tumor reaches During 4-5mm, the blocking VEGF signal path of first time low dosage (the 1/4 of the general dosage of mouse) is carried out to mouse The treatment processing (being set to 0 day) of medicine (DC101) and rabbit anti-mouse igg monoclonal antibody (control), drug injection frequency For once every three days, dosage is 10mg/kg (being only the 1/4 of the general dosage of mouse), is administered 4 times altogether. The paraformaldehyde for carrying out 4% for 11 days to mouse is irrigated, and prepares tumor tissues frozen section (20 μm), and to this Histotomy carries out the mark dye of endothelial cell marker thing CD31 molecules and perithelial cells label NG2 molecules Color.Tumor region, which is randomly selected, using confocal laser scanning microscope, CLSM carries out SABC IMAQ (often Individual 4-6 region of tumor tissues, every group of 6-8 tumour), multiplication factor 20 ×.Statistical analysis tumor vessel Density, pericyte coverage rate.
As a result as shown in figure 1, low dosage anti-angiogenic medicaments DC101 (10mg/kg) treatment after the 11st My god, tumor vascular density still can be observed and substantially reduces (Figure 1A and Figure 1B), and pericyte coverage rate is notable Increase (Figure 1A and Fig. 1 C).As a result show that low dosage anti-angiogenic medicaments can induce long-acting tumor vessel just Normalizing.
Embodiment 2
The anti-angiogenic medicaments combined vaccine of low dosage treats the influence to mouse tumor
In order to study influence of the tumor vessel normalization to immunotherapy of tumors, the anti-angiogenic of low dosage has been carried out Generate medication combined vaccine therapy experiment.When breast cancer mouse tumor model MCaP0008 diameter of tumor reaches During 3mm, random packet, intraperitoneal injection 5 × 106Mitomycin pretreatment MCaP0008 tumor cell vaccines or It is isometric PBS (control), injection time is respectively the 7th, 9,12,14 day.Respectively the 13rd, 16, 19,20 days injection anti-angiogenic medicaments DC101 (10mg/kg) or rabbit anti-mouse igg (10mg/kg).From the 13rd It starts to measure tumor size, and measurement in every three days is once.Draw tumor growth curve.Every group of 10 mouse.
As a result as shown in Fig. 2 individually vaccine therapy is not obvious to the inhibition of mice tumors grew, and The tumor vessel normalization of the anti-angiogenic medicaments induction of low dosage can significantly increase immunotherapy of tumors Effect.As a result show, compared with the anti-angiogenic medicaments treatment of single vaccine therapy and low dosage, The anti-angiogenic medicaments combined vaccine treatment of low dosage can significantly inhibit mice tumors grew.
Embodiment 3
The anti-angiogenic medicaments of low dosage treat the tumor-infiltrated influence to T cell
Strengthen the mechanism of immunization therapy to study tumor vessel normalization, to the CD8+ infiltrated in tumor tissues Analyzed with CD4+T lymphocytes.When MCaP0008 breast cancer mouse diameter of tumor reaches 4-5mm, The DC101 of low dosage and the treatment processing of tumor cell vaccine, DC101 and control rabbit-anti mouse are carried out to mouse IgG drug injection frequency is administered 4 times altogether for once every three days, dosage is 10mg/kg.Then collect swollen Tumor tissue, prepares single cell suspension, and then analyze tumor-infiltrated CD4+T cells using stream type cell analyzer With CD8+T cells.
As a result as shown in figure 3, single low dosage blocking VEGF signal path medicine (group：PBS/D10) can be with The wetting capacity of CD8+ and CD4+ lymphocytes is significantly improved, and low dosage blocking VEGF signal path medicine joins Close vaccine therapy (group：Vaccine/D10) after, the wetting capacity of CD4+ lymphocytes is further significantly increased. As a result show, the anti-angiogenic medicaments treatment of low dosage can promote the tumor-infiltrated of T cell.
Embodiment 4
Influence of the anti-angiogenic medicaments of routine dose to tumor vaccine therapy
Experiment condition is same as Example 2, and difference is that the dosage for the anti-angiogenic medicaments applied is 40mg/kg, i.e., using the anti-angiogenic medicaments of routine dose, study its influence to immunotherapy of tumors.
As a result as shown in figure 4, individually vaccine therapy is not obvious to the inhibition of mice tumors grew.Together When, the inhibition of the therapeutic alliances of the anti-angiogenic medicaments and vaccine of routine dose to mice tumors grew There is no notable difference with the therapeutic effect of single routine dose anti-angiogenic medicaments.The above results show often Advise the anti-angiogenic medicaments of dosage (high dose), it is impossible to improve the therapeutic effect of tumor vaccine.
Embodiment 5
The pretreatment of blood vessel normalization combines the CAR-T clinical treatments of PCI
CAR-T cell therapies are that the very promising new cellular immunity that can effectively treat tumour is treated Method.Receive the complete remission rate of the relapsed or stubborn Patients With Acute Lymphoblastic Leukemia of CD19CAR-T treatments Reach 90%.But, performance of the CAR-T cells in treatment of solid tumors is not satisfactory, in the world Trial for solid tumor CAR-T cell therapies is also and failed.Tumor tissues internal blood vessel skewness, The factors such as inside tumor oxygen supply deficiency seriously hinder CAR-T cell systems to feed back the treatment that scheme is directed to solid tumor Inside validity, the infiltration that the CAR-T cells for causing system to feed back can not be fully effective to tumor tissues, and Cause CAR-T cells can not inside tumor by fully effective activation, propagation and retain for a long time. Therefore, in order to improve the adverse effect that the particularity of above-mentioned three point entities knurl is caused to CAR-T cell therapies, Intervention strategies are selected by inside CAR-T cell infusions to tumor tissues, while coordinating low dosage (1/5 and 1/10 Normal dose) Avastin (bevacizumab, anti-angiogenic medicaments), induced tumor blood vessel normalization.
After being agreed to by the examination ＆ approval of clinical hospitals Ethics Committee, to 1 MUC1+ that endorsed informed consent form Patients with advanced solid tumors's (late period adenocarcinoma of seminal vesicle) has carried out clinical research experiment.Patient is injected intravenously first The blocking of 1mg/kg (the 1/5 of normal dose, the dosage is obtained according to above-mentioned mouse test results presumption) dosage VEGF signal path medicine Avastin (bevacizumab, targeting ligand VEGF), the equal agent of duplicate injection after 2 weeks Blocking VEGF signal path medicine is measured, the tumor focus then shifted after 1 week to the patient is under color ultrasound guidance The intratumor injection of MUC1CAR-T cells is carried out.After CAR-T cell infusions 1 week, blocking VEGF is injected again Signal path medicine, but dosage be reduced to 0.5mg/kg (the 1/10 of normal dose, the dosage according to more than it is small Mouse result of the test, which speculates, to be obtained).After CAR-T cell infusions 3 weeks, tumor tissues are won in operation, are fabricated to stone Wax is cut into slices, and carries out HE dyeing.
As a result, can be with as shown in figure 5, the pink oval tissue of color is fresh downright bad tumour cell in figure See the crack that postnecrotic tumour cell is formed with perienchyma.As a result show, the blocking VEGF of low dosage The tumor locus bleeding tumor that signal path medicine combines CAR-T cell infusions with MUC1CAR-T is substantially downright bad, Show good therapeutic effect.
All documents referred in the present invention are all incorporated as reference in this application, just as each document It is individually recited as with reference to such.In addition, it is to be understood that after the above-mentioned instruction content of the present invention has been read, Those skilled in the art can make various changes or modifications to the present invention, and these equivalent form of values equally fall within this Shen Please appended claims limited range.

Claims (10)

1. a kind of pharmaceutical composition for being used to treat entity tumor, it is characterised in that described pharmaceutical composition is included First active component and the second active component, wherein, the first active component is VEGF signal path blocking agents, second Active component is immunotherapy of tumors agent,
And described pharmaceutical composition is unit dosage form, and the first active component contains described in each unit dosage form 0.1 to 0.5 (the preferably 0.15-0.4, or 0.2-0.25) for routine dose is measured, wherein, the conventional agent Measure as 200-400mg/50kg body weight.
2. pharmaceutical composition as claimed in claim 1, it is characterised in that in a described unit dosage form The content of first active component correspond to the 0.1 of the total routine dose of the first active component being administered every time to 0.5。
3. pharmaceutical composition as claimed in claim 1, it is characterised in that described VEGF signal paths resistance Disconnected agent includes the antibody and tyrosine kinase inhibitor of blocking VEGF signal path.
4. pharmaceutical composition as claimed in claim 1, it is characterised in that described blocking VEGF signal leads to The antibody on road includes：Thunder not Lu Dankang, bevacizumab or its combination.
5. pharmaceutical composition as claimed in claim 1, it is characterised in that first active component and Two active components are mixing or individualism.
6. pharmaceutical composition as claimed in claim 1, it is characterised in that described the first active component and The total content of second active component is 70~100wt%, by the gross weight of active component in described pharmaceutical composition Meter.
7. pharmaceutical composition as claimed in claim 1, it is characterised in that described immunotherapy of tumors bag Include CAR-T cell therapies, CAR-NK cell therapies, tumor vaccine therapy, immunity inspection point inhibitor for treating or It is combined.
8. the purposes of the VEGF signal path blocking agents of a kind of low dosage, it is characterised in that for preparing a medicine Composition or medicine box, described pharmaceutical composition or medicine box are used for the curative effect for improving immunotherapy of tumors,
Wherein, low dosage refers to the dosage A1 and conventional administration dosage A0 of the VEGF signal paths blocking agent Compare, A1≤1/2A0.
9. purposes as claimed in claim 8, it is characterised in that described medicine box includes 2-4 unit dosage form, The content of the antibody of VEGF signal path blocking agents in each unit formulation is the 0.1 to 0.5 of routine dose, Wherein, the routine dose is 200-400mg/50kg body weight.
10. a kind of medicine box for being used to improve immunotherapy of tumors effect, it is characterised in that the medicine box includes：
(a) the first pharmaceutical composition, first pharmaceutical composition includes VEGF signal path blocking agents, and pharmacy Upper acceptable carrier, and first pharmaceutical composition is unit dosage form, and the medicine box includes 2-4 Individual unit dosage form, wherein the content of the VEGF signal path blocking agents in the unit dosage form is the 0.1 of routine dose To 0.5 (preferably 0.15-0.4, or 0.2-0.25), wherein, the routine dose is 200-400mg/50kg Body weight；
(b) the second optional pharmaceutical composition, the second described pharmaceutical composition is immunotherapy of tumors agent；
(c) specification.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103285392A (en) * 2005-04-26 2013-09-11 卫材R&D管理株式会社 Compositions for cancer immunotherapy and use thereof
CN104271601A (en) * 2012-05-31 2015-01-07 霍夫曼-拉罗奇有限公司 Methods of treating cancer using PD-l1 axis binding antagonists and VEGF antagonists
CN104436194A (en) * 2013-09-18 2015-03-25 北京大学 Anti-cancer composition with synergistic effect
WO2015119930A1 (en) * 2014-02-04 2015-08-13 Pfizer Inc. Combination of a pd-1 antagonist and a vegfr inhibitor for treating cancer

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103285392A (en) * 2005-04-26 2013-09-11 卫材R&D管理株式会社 Compositions for cancer immunotherapy and use thereof
CN104271601A (en) * 2012-05-31 2015-01-07 霍夫曼-拉罗奇有限公司 Methods of treating cancer using PD-l1 axis binding antagonists and VEGF antagonists
CN104436194A (en) * 2013-09-18 2015-03-25 北京大学 Anti-cancer composition with synergistic effect
WO2015119930A1 (en) * 2014-02-04 2015-08-13 Pfizer Inc. Combination of a pd-1 antagonist and a vegfr inhibitor for treating cancer

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
应杰儿等: "不同剂量贝伐单抗联合伊立替康对荷人结肠癌DLD-1裸鼠皮下移植瘤生长的影响", 《肿瘤学杂志》 *
戴明等: "贝伐单抗对荷耐顺铂人肺腺癌A549/DDP裸鼠皮下移植瘤生长的影响", 《南方医科大学学报》 *
陆萌等: "活体观察贝伐单抗对人骨肉瘤裸鼠移植瘤模型的作用", 《临床肿瘤学杂志》 *
陈俊青等: "贝伐单抗与重组人血管内皮抑素对人乳腺癌MCF-7 细胞裸鼠移植瘤生长的影响", 《中国肿瘤》 *

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