CN108310377A - It is a kind of that PD-1 antibody is combined to the application for being used to prepare anticancer drug with melbine - Google Patents
It is a kind of that PD-1 antibody is combined to the application for being used to prepare anticancer drug with melbine Download PDFInfo
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- CN108310377A CN108310377A CN201810204275.5A CN201810204275A CN108310377A CN 108310377 A CN108310377 A CN 108310377A CN 201810204275 A CN201810204275 A CN 201810204275A CN 108310377 A CN108310377 A CN 108310377A
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- Prior art keywords
- melbine
- antibody
- cancer
- anticancer drug
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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
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/13—Amines
- A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
Abstract
The invention discloses it is a kind of 1 antibody of PD is combined with melbine be used to prepare the application of anticancer drug, be related to pharmaceutical technology field.1 antibody of PD is combined with melbine is used to prepare anticancer drug；The anticancer drug includes 1 antibody of PD and melbine；The content of melbine is 0.02mg~0.10mg in the anticancer drug 0.1ml；The content of 1 antibody of PD is 0.01mg~0.05mg in the anticancer drug 0.1ml.The present invention is used to prepare the effect of anticancer drug by combining 1 antibody of PD with melbine, and monoclonal antibody treatment, the imperfect problem of cancer resistant effect can only be carried out by solving existing technology.
Description
Technical field
The invention belongs to pharmaceutical technology field, more particularly to it is a kind of PD-1 antibody is combined with melbine be used to prepare
The application of anticancer drug.
Background technology
1 (ProgrammedDeath-1 of programmed death receptor；PD-1), it is a kind of important immunosuppression molecule, is
CD28 superfamily members are initially to clone to come from the mouse Tcell hybridoma 2B4.11 of apoptosis.PD-1 is mainly being activated
T cell and B cell in express, have the function of inhibit cell-stimulating, this is a kind of normal homeostasis in immune system,
Because excessive T/B cell-stimulatings can cause autoimmune disease, PD-1 to be equivalent to one of guardrail in human body.
In tumor microenvironment, tumour cell can high ligand PD-L1 (apoptosis-ligand 1 or the table for expressing PD-1
Face antigen differentiation cluster 274 or B7 autoploids).The PD-1 of the PD-L1 and T cell surface that are expressed on tumour cell interact, and lead
PD-1 access sustained activations in tumor microenvironment, T cell function is caused to be suppressed, it can not killing tumor cell.PD-1 monoclonals are anti-
Body can be by blocking this access, and the function of T cell is restored in part, so using the T cell of activation and corresponding cell because
Sub- killing tumor cell, to which it becomes the critical treatment means of immunotherapy for cancer.
Melbine is a line oral hypoglycemic agents of current clinical treatment diabetes B.In recent years, has multinomial research
Experiment shows that melbine has a degree of inhibiting effect to kinds of tumors.Relevant clinical research is also shown, and is taken
The related mortality for the risk and tumour that the diabetic of melbine suffers from malignant tumour is significantly less than using other drops
The diabetic of sugared drug or insulin therapy, to show that its anticancer effect is unrelated with blood sugar decreasing effect.
There are following defects in anticancer drug at present：The prior art can only carry out monoclonal antibody treatment, and cancer resistant effect is not
Enough good problems.So far it PD-1 antibody is combined with melbine is used to prepare anticancer drug there has been no relevant report.By PD-1
Antibody is combined with melbine has far-reaching clinical meaning for treating cancer.
Invention content
The purpose of the present invention is to provide it is a kind of PD-1 antibody is combined with melbine be used to prepare answering for anticancer drug
With being used to prepare the effect of anticancer drug by combining PD-1 antibody with melbine, solve existing technology only
It can carry out monoclonal antibody treatment, the imperfect problem of cancer resistant effect.
In order to solve the above technical problems, the present invention is achieved by the following technical solutions：
PD-1 antibody being combined the application for being used to prepare anticancer drug by the present invention to be a kind of with melbine, and the PD-1 is anti-
Body is combined with melbine is used to prepare anticancer drug；The anticancer drug includes PD-1 antibody and melbine；The anticancer
The content of melbine is 0.02mg~0.10mg in drug 0.1ml；The content of PD-1 antibody is in the anticancer drug 0.1ml
0.01mg~0.05mg.
Further, the quality proportioning of the PD-1 antibody and melbine is 1：2~10；The melbine is treatment
One line oral medicine of diabetes B has certain anticancer effect, it may have kill the function of tumor stem cell；The PD-1
Monoclonal antibody has lethal effect to a variety of cancer cells；The melbine can enhance PD-1 monoclonal antibodies to cancer
Curative effect.
Further, the application of the anticancer drug be treatment cancer of pancreas, it is lung cancer, gastric cancer, liver cancer, colorectal cancer, black
Melanoma, nephroncus, oophoroma, prostate cancer, carcinoma of urinary bladder, breast cancer, the cancer of the esophagus, colorectal cancer, nasopharyngeal carcinoma, brain tumor, cervical carcinoma,
The drug of leukemia, osteocarcinoma, lymph cancer or other cancers.
The invention has the advantages that：
The present invention is used to prepare the effect of anticancer drug, wherein melbine by combining PD-1 antibody with melbine
It is the line oral medicine for treating diabetes B, there is certain anticancer effect, it may have kill the function of tumor stem cell；PD-
1 monoclonal antibody has lethal effect to a variety of cancer cells；PD-1 monoclonal antibodies can be made to cancer with melbine
The effect of more preferably the advantages of.
Certainly, it implements any of the products of the present invention and does not necessarily require achieving all the advantages described above at the same time.
Specific implementation mode
Technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is only
It is a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, ordinary skill people
All other embodiment that member is obtained without creative efforts, shall fall within the protection scope of the present invention.
Embodiment one
The present invention be it is a kind of PD-1 antibody combined to the application for being used to prepare anticancer drug with melbine, PD-1 antibody and
Melbine, which is combined, is used to prepare anticancer drug；Anticancer drug includes PD-1 antibody and melbine；Two in anticancer drug 0.1ml
The content of first biguanides is 0.02mg~0.10mg；The content of PD-1 antibody is 0.01mg~0.05mg in anticancer drug 0.1ml.
Wherein, the quality proportioning of PD-1 antibody and melbine is 1：2~10；Melbine is treatment diabetes B
One line oral medicine has certain anticancer effect, it may have kill the function of tumor stem cell；PD-1 monoclonal antibodies are to a variety of
Cancer cell has lethal effect；Melbine can enhance the effect of PD-1 monoclonal antibodies are to cancer.
Wherein, the application of anticancer drug is treatment cancer of pancreas, lung cancer, gastric cancer, liver cancer, colorectal cancer, melanoma, kidney
Tumor, oophoroma, prostate cancer, carcinoma of urinary bladder, breast cancer, the cancer of the esophagus, colorectal cancer, nasopharyngeal carcinoma, brain tumor, cervical carcinoma, leukemia, bone
The drug of cancer, lymph cancer or other cancers.
Wherein, the increment that can inhibit pancreatic cancer cell is used in combination with melbine in PD-1 antibody：
Test material therefor：Pancreas cancer cell strain panc-l；Melbine, after the dissolving of 10%FBS+90%DMEM culture mediums
Packing, 4 DEG C of preservations；PD-1 antibody dispenses after the dissolving of 10%FBS+90%DMEM culture mediums, 4 DEG C of preservations；CCK8 cell proliferations with
Activity determination；10%FBS+90%DMEM culture mediums；This experiment agents useful for same is market common agents.
Cell experiment flow is as follows：
(1) panc-l cells in good condition are taken, 2.0 × 104/mL that density is are adjusted, by cell suspension inoculation in 96
Orifice plate (holes 0.1mL/) (setting blank assay, only add culture medium)；
(2) adherent growth 48 hours discard original culture medium；Culture medium (0.1mL) is added in control group, and experimental group is added two
First biguanides (0.1mL melbine contents 0.02mg, 0.03mg, 0.04mg, 0.05mg, 0.06mg), it is anti-that PD-1 is added in experimental group
Body (0.1mL PD-1 antibody content 0.01mg), melbine is added in experimental group, and (0.1mL diformazans are double with the antibody combined uses of PD-1
Guanidine content 0.02mg, 0.03mg, 0.04mg, 0.05mg, 0.06mg and PD-1 antibody content 0.01mg), blank assay group is changed
Culture medium is cultivated 48 hours；
(3) the porose culture medium of institute is discarded, CCK8 solution (10uL) is added in hole, continues culture 2 hours；
(4) its absorbance is surveyed at 450nm wavelength with microplate reader；
(5) cell viability is calculated；Cell viability=(A experiment-A blank)/(A control-A blank).
Experimental result：
The experimental result that table 1panc-l pancreas cancer cell strains are detected by cell culture, drug-treated, CCK8：
As a result it shows：Panc-l pancreas cancer cell strains pass through medium treatment survival rate 100%, by melbine
It is 90%, 88%, 83%, 70%, 89% that (0.02mg, 0.03mg, 0.04mg, 0.05mg, 0.06mg), which handles survival rate, is passed through
PD-1 antibody handle survival rate be 59%, by melbine (0.02mg, 0.03mg, 0.04mg, 0.05mg, 0.06mg) with
The antibody combined use survival rates of PD-1 are 59%, 60%, 55%, 40%, 57%；With 48 hours survival rates of medium treatment
100% compares, and melbine (0.02mg, 0.03mg, 0.04mg, 0.06mg) difference is statistically significant (P ＜ 0.05), diformazan
Biguanides (0.05mg) difference is statistically significant (P ＜ 0.05)；Compared with PD-1 antibody individually handles survival rate 59%, diformazan is double
Guanidine (0.02mg, 0.03mg, 0.04mg, 0.06mg) is antibody combined statistically significant (P ＜ 0.05) using difference with PD-1, and two
First biguanides (0.05mg) is antibody combined statistically significant (P ＜ 0.05) using difference with PD-1；The diformazan of comprehensive different amounts
Biguanides individually processing and with the antibody combined experimental results used of PD-1, mass ratio 5 may be selected:1 is ratio used in subsequent experimental.
Embodiment two
The present invention be it is a kind of PD-1 antibody combined to the application for being used to prepare anticancer drug with melbine, PD-1 antibody and
Melbine, which is combined, is used to prepare anticancer drug；Anticancer drug includes PD-1 antibody and melbine；Two in anticancer drug 0.1ml
The content of first biguanides is 0.02mg~0.10mg；The content of PD-1 antibody is 0.01mg~0.05mg in anticancer drug 0.1ml.
Wherein, the quality proportioning of PD-1 antibody and melbine is 1：2~10；Melbine is treatment diabetes B
One line oral medicine has certain anticancer effect, it may have kill the function of tumor stem cell；PD-1 monoclonal antibodies are to a variety of
Cancer cell has lethal effect；Melbine can enhance the effect of PD-1 monoclonal antibodies are to cancer.
Wherein, the application of anticancer drug is treatment cancer of pancreas, lung cancer, gastric cancer, liver cancer, colorectal cancer, melanoma, kidney
Tumor, oophoroma, prostate cancer, carcinoma of urinary bladder, breast cancer, the cancer of the esophagus, colorectal cancer, nasopharyngeal carcinoma, brain tumor, cervical carcinoma, leukemia, bone
The drug of cancer, lymph cancer or other cancers.
Wherein, the increment that can significantly inhibit pancreatic cancer cell is used in combination with melbine in PD-1 antibody：
Test material therefor：Pancreas cancer cell strain panc-l, MIAPACA-2, BXPC-3；Melbine, 10%FBS+
It is dispensed after the dissolving of 90%DMEM culture mediums, 4 DEG C of preservations；PD-1 antibody dispenses, 4 after the dissolving of 10%FBS+90%DMEM culture mediums
DEG C preserve；CCK8 cell proliferations and Activity determination；10%FBS+90%DMEM culture mediums；This experiment agents useful for same is that market is normal
See reagent.
Cell experiment flow is as follows：
(1) cell in good condition is taken, 2.0 × 104/mL that density is is adjusted, by cell suspension inoculation in 96 orifice plates
(holes 0.1mL/) (setting blank assay, only add culture medium)；
(2) adherent growth 48 hours discard original culture medium；It is separately added into after grouping：Control group only culture medium (0.1mL/
Hole), experimental group melbine (0.1mL melbine content 0.05mg), experimental group PD-1 antibody (0.1mL PD-1 contents
0.01mg), experimental group melbine and antibody combined use (0.1mL melbine content 0.05mg and the PD-1 contents of PD-1
0.01mg), blank assay group change culture medium can (holes 0.1mL/), cultivate 48 hours；
(3) the porose culture medium of institute is discarded, CCK8 solution (10ul) is added in hole, continues culture 2 hours；
(4) its absorbance is surveyed at 450nm wavelength with microplate reader；
(5) cell viability is calculated；Cell viability=(A experiment-A blank)/(A control-A blank).
Experimental result：
The experimental result that the different pancreas cancer cell strains of table 2 are detected by cell culture, drug-treated, CCK8：
As a result it shows：
The survival rate that 48 hours panc-l cell strains are handled by 4 kinds of distinct methods is respectively 100%, 71%, 57%,
43%；
The survival rate that 48 hours MIAPACA-2 cell strains are handled by 4 kinds of distinct methods is respectively 100%, 73%,
60%, 38%；
The survival rate that 48 hours BXPC-3 cell strains are handled by 4 kinds of distinct methods is respectively 100%, 74%, 58%,
40%；
By 3 groups of difference cell line experiments interpretations of result, melbine and PD-1 are anti-compared with 100% Cell viability of control group
Body is used in combination group and is statistically significant (P ＜ 0.01), and PD-1 antibody groups are used alone and melbine group is used alone
Experimental result and control group difference be respectively less than be used in combination group difference it is statistically significant (P ＜ 0.05)；Illustrate that diformazan is double
Guanidine and the antibody combined uses of PD-1 are more preferable than melbine, PD-1 antibody exclusive use cancer resistant effect.
Embodiment three
The present invention be it is a kind of PD-1 antibody combined to the application for being used to prepare anticancer drug with melbine, PD-1 antibody and
Melbine, which is combined, is used to prepare anticancer drug；Anticancer drug includes PD-1 antibody and melbine；Two in anticancer drug 0.1ml
The content of first biguanides is 0.02mg~0.10mg；The content of PD-1 antibody is 0.01mg~0.05mg in anticancer drug 0.1ml.
Wherein, the quality proportioning of PD-1 antibody and melbine is 1：2~10；Melbine is treatment diabetes B
One line oral medicine has certain anticancer effect, it may have kill the function of tumor stem cell；PD-1 monoclonal antibodies are to a variety of
Cancer cell has lethal effect；Melbine can enhance the effect of PD-1 monoclonal antibodies are to cancer.
Wherein, the application of anticancer drug is treatment cancer of pancreas, lung cancer, gastric cancer, liver cancer, colorectal cancer, melanoma, kidney
Tumor, oophoroma, prostate cancer, carcinoma of urinary bladder, breast cancer, the cancer of the esophagus, colorectal cancer, nasopharyngeal carcinoma, brain tumor, cervical carcinoma, leukemia, bone
The drug of cancer, lymph cancer or other cancers.
Wherein, the growth that can preferably inhibit entity tumor in nude mouse is used in combination with melbine in PD-1 antibody：
Test material therefor：Several males of normal 6 week old nude mice, every mouse body are taken to weigh about as 20g；Panc-l cells
Strain；Melbine dispenses, 4 DEG C of preservations after sterile PBS solution dissolving；PD-1 antibody dispenses, 4 DEG C after sterile PBS solution dissolving
It preserves.
Mouse experiment flow is as follows：
(1) foundation of mouse pancreas cancer model uses subcutaneous transplantation knurl model, convenient for observation tumour growth situation；Take cell
Density is the panc-l cells of 2.0 × 107/mL, and disinfection mouse is intended to the skin of injection site, in every nude mice dorsal sc injection
0.1mL cell suspensions are inoculated with out after 10d when growing 2-3mm tumors, mouse pancreas cancer modelling success；
(2) successful mouse will be modeled and be randomly divided into 4 groups, PBS control group, melbine experimental group, PD-1 antibody assays
Group, melbine and PD-1 it is antibody combined use experimental group；Sterile PBS is administered in control group after nude inoculation tumour cell 10 days
Melbine (intraperitoneal injection 0.1ml 10mg/kg/d), (intraperitoneal injection of PD-1 antibody is administered in (intraperitoneal injection 0.1mL), experimental group
0.1ml 2mg/kg/d), melbine and PD-1 it is antibody combined use (intraperitoneal injection 0.1ml melbine 10mg/kg/d with
PD-1 antibody 2mg/kg/d), control group and the equal successive administration of experimental group 12 days；
(3) it is discontinued 24 hours after weighing and puts to death mouse, remove subcutaneous transplantation tumor, weigh and preserve；It is calculated by average knurl weight
Tumour inhibiting rate；Tumour inhibiting rate=[(control group gross tumor volume-experimental group gross tumor volume)/control group gross tumor volume] × 100%.
Mouse test results：
The results show that the tumour inhibiting rate that experimental group is used in combination with melbine for PD-1 antibody is apparently higher than PBS control group, it is poor
Different statistically significant (P ＜ 0.05), the tumour inhibiting rate that experimental group is used in combination with melbine for PD-1 antibody are higher than melbine
Experimental group and PD-1 antibody assay groups, difference are statistically significant (P ＜ 0.05).
In the description of this specification, the description of reference term " one embodiment ", " example ", " specific example " etc. means
Particular features, structures, materials, or characteristics described in conjunction with this embodiment or example are contained at least one implementation of the present invention
In example or example.In the present specification, schematic expression of the above terms may not refer to the same embodiment or example.
Moreover, particular features, structures, materials, or characteristics described can be in any one or more of the embodiments or examples to close
Suitable mode combines.
Present invention disclosed above preferred embodiment is only intended to help to illustrate the present invention.There is no detailed for preferred embodiment
All details are described, are not limited the invention to the specific embodiments described.Obviously, according to the content of this specification,
It can make many modifications and variations.These embodiments are chosen and specifically described to this specification, is in order to preferably explain the present invention
Principle and practical application, to enable skilled artisan to be best understood by and utilize the present invention.The present invention is only
It is limited by claims and its full scope and equivalent.
Claims (3)
1. a kind of combining PD-1 antibody with melbine the application for being used to prepare anticancer drug, it is characterised in that：The PD-1
Antibody is combined with melbine is used to prepare anticancer drug；The anticancer drug includes PD-1 antibody and melbine；It is described anti-
The content of melbine is 0.02mg~0.10mg in cancer drug 0.1ml；The content of PD-1 antibody in the anticancer drug 0.1ml
For 0.01mg~0.05mg.
2. it is according to claim 1 it is a kind of PD-1 antibody is combined to the application for being used to prepare anticancer drug with melbine,
It is characterized in that, the quality proportioning of the PD-1 antibody and melbine is 1：2~10.
3. it is according to claim 1 it is a kind of PD-1 antibody is combined to the application for being used to prepare anticancer drug with melbine,
It is characterized in that, the application of the anticancer drug be treatment cancer of pancreas, lung cancer, gastric cancer, liver cancer, colorectal cancer, melanoma,
Nephroncus, oophoroma, prostate cancer, carcinoma of urinary bladder, breast cancer, the cancer of the esophagus, colorectal cancer, nasopharyngeal carcinoma, brain tumor, cervical carcinoma, leukemia, bone
The drug of cancer, lymph cancer or other cancers.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112791182A (en) * | 2020-12-31 | 2021-05-14 | 中山大学 | Application of pharmaceutical composition of metformin and anti-PD-1 antibody in preparation of liver cancer drugs |
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| CN104903450A (en) * | 2012-11-05 | 2015-09-09 | 普隆奈治疗公司 | Dosing and administration of oligonucleotide cancer therapies |
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