CN106943400B - Enhance liver cancer cells to the compound and its preparation of CIK cell killing sensibility - Google Patents
Enhance liver cancer cells to the compound and its preparation of CIK cell killing sensibility Download PDFInfo
- Publication number
- CN106943400B CN106943400B CN201710290463.XA CN201710290463A CN106943400B CN 106943400 B CN106943400 B CN 106943400B CN 201710290463 A CN201710290463 A CN 201710290463A CN 106943400 B CN106943400 B CN 106943400B
- Authority
- CN
- China
- Prior art keywords
- cell
- compound
- liver cancer
- sensibility
- cik cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of enhancing liver cancer cells to the compound and its preparation of CIK cell killing sensibility, specifically provides a kind of application of pyridine-imidazole class organic compound in drug of the preparation enhancing liver cancer cells to CIK cell killing sensibility.Present invention demonstrates that pyridine-imidazole class compound, which can effectively enhance liver cancer cells, kills sensibility to CIK cell, it can be prepared into pharmaceutically acceptable pharmaceutical preparation, just play preferably immunotherapeutic effects using a small amount of CIK cell to realize.
Description
Technical field
The invention belongs to immunotherapy fields, are related to CIK cell, and in particular to a kind of enhancing liver cancer cells are to CIK cell
Kill the compound and its preparation of sensibility.
Background technique
Primary carcinoma of liver (hereinafter referred to as liver cancer) is one of most common malignant tumour in world wide, every year about 63
Ten thousand new cases, and disease incidence is on the rise in recent years.The course of disease of liver cancer patient is short, clinically more difficult early detection,
Often the state of an illness has entered middle and advanced stage when making a definite diagnosis, and case fatality rate is high.The treatment of liver cancer at present is mainly with the excision, liver transfer operation, office of performing the operation
Portion's ablation, chemotherapy embolism and other locoregional treatments, molecular targeted therapy etc., but the therapeutic effect of these methods is not
Ideal, because the innate immunity, specific immune function are low in liver cancer patient body, radical surgery resection rate is low, multiple after operation
Hair rate is high, radiation, chemotherapy weak curative effect, sends out in Yi Fasheng liver and extrahepatic metastases, therefore is badly in need of finding new effective ways
To treat liver cancer.Biological therapy has become the 4th kind of mode in combined therapy of tumour, is increasingly taken seriously.
Immunocompetent cell currently used for tumor biotherapy mainly has tumor infiltrating lymphocyte (TIL), dendron
Shape cell (DC) and cytokine induced kill cell (CIK) etc..CIK cell is a kind of novel immunocompetent cell,
The surface marker of its main effects cell is CD3+CD56+, compared with other adoptive immunotherapy cells, has growth rate
Fastly, it kills tumor activity height, kill the advantages that tumor spectrum is wide.Studies have shown that adoptive cellular immunotherapy to inhibit liver cancer recurrence and transfer,
Raising patient's cure rate, improvement patients ' life quality, extension life cycle all play a significant role.
How with less CIK cell the important research that preferably immunotherapeutic effects are researchers is just played
Direction.
Summary of the invention
The purpose of the present invention is to provide a kind of enhancing liver cancer cells to the compound and its system of CIK cell killing sensibility
Agent, to use less CIK cell just to play preferably immunotherapeutic effects.
Realize that above-mentioned purpose technical solution of the present invention is as follows:
A kind of pyridine-imidazole class organic compound kills the drug of sensibility in preparation enhancing liver cancer cells to CIK cell
In application, the pyridine-imidazole class organic compound have following general formula structure:
Wherein, R2For-OCH2CH3Or-OH；
R1For-CH2Or-CH (CH3)2Or-CH2CH2CH2-。
Preferably, pyridine-imidazole class organic compound is selected from following compounds:
A kind of pharmaceutical preparation killing sensibility to CIK cell for enhancing liver cancer cells, including above-mentioned pyridine-imidazole
Class organic compound further includes pharmaceutically acceptable carrier or excipient, pharmaceutically acceptable dosage form is made.
Preferably, the pharmaceutically acceptable carrier or excipient include one or more solids, semisolid or liquid
Body auxiliary material.
Preferably, the pharmaceutically acceptable dosage form includes tablet, capsule, granule, injection, pill, dissipates
Agent, paste, oral liquid.
Outstanding advantages of the invention:
Present invention demonstrates that pyridine-imidazole class compound, which can effectively enhance liver cancer cells, kills sensibility to CIK cell,
It can be prepared into pharmaceutically acceptable pharmaceutical preparation, just play preferably immune control using a small amount of CIK cell to realize
Therapeutic effect.
Detailed description of the invention
Fig. 1 is that each group CIK cell compares the killing rate of HepG2.
Specific embodiment
Just specifically introduce essentiality content of the invention in conjunction with the embodiments below, due to length, experimentation is retouched
Stating can not accomplish very in detail, and the part being not described in detail in all experiments is conventional behaviour well known to those skilled in the art
Make.
One, experimental material
The pyridine-imidazole class organic compound of number CM1-6 is known compound, synthesizes personnel's reference by our company
Document and the synthesis of organic synthesis conventional method, and confirmed through mass spectrum and nuclear-magnetism.
Human hepatoma cell line HepG2 is our company's long-term preservation.Recombinant human interferon alpha 2 rhIFN- γ, combining human interleukins
Plain rhIL-2, CD3 mouse anti-human monoclonal antibody, CIK cell separating liquid, PBS, serum free medium MD-CM-STMN,
RPMI1640 culture medium, fetal calf serum FBS, MTT, DMSO, pancreatin are purchased from Nanjing and build up biology.
Two, experimental method
1, prepared by peripheral blood mononuclear cells
Aseptic aspiration healthy human peripheral blood about 50mL, anticoagulant heparin (heparin 15IU/mL) dilute periphery with isometric PBS
Blood.It is slowly added into the 50mL centrifuge tube containing cell separating liquid by the volume ratio of 1:1,2000r/min is centrifuged 20min.It takes
More intermediate annular milky mononuclearcell layer moves into 50mL centrifuge tube, and appropriate PBS is added, and 1800r/min is centrifuged 8min, abandons
Supernatant washs 2 times, cell count.
2, CIK cell Fiber differentiation
It is 1 × 10 with the MD-CM-STMN cell culture medium adjustment mononuclearcell density containing 5%FBS6A/mL, connects
20mL is in culture bottle for kind, and total number of cells are 2 × 107, the addition of (the 0th day) same day rhIL-21000U/mL, rhIFN- γ
1000U/mL, CD3 mouse anti-human monoclonal antibody 5mg/L place 37 DEG C, 5%CO2Start to cultivate in incubator.Start to mend within 3rd day
Adding MD-CM-STMN cell culture medium, while supplementing CD3 mouse anti-human monoclonal antibody, rhIFN- γ, rhIL-2, dosage is the same,
Hereafter every 3d adds fresh culture, adds rhIFN- γ, dosage is identical, and rhIL-2 dosage halves as 500U/mL.
3, CIK cell proliferation assay
Trypan Blue method counts cell on the 0th, 5,8,13,20 day with cell counting board respectively at culture, weight
It answers 3 times and is averaged.
4, the culture of tumor cell line
By HepG2 cell inoculation in culture bottle, the RPMI 1640 containing 10%FBS is added, attached cell is with 0.25%
Pancreatin digestion.The cell of logarithmic growth phase is inoculated in 96 orifice plates, carries out killing experiments in vitro.
5, cytotoxicity of the mtt assay measurement CM 1-6 compound to HepG2 cell
Logarithmic growth phase HepG2 cell, adjustment cell suspension density are 5 × 103A/mL is inoculated in 96 orifice plates, every hole
200μL.Liquid is changed after cell is adherent, administration group is separately added into 2 μM of compound CM 1-6, and culture solution is added in control group, and every group equal
If 3 parallel holes, are placed on 37 DEG C, 5%CO2After acting on 48h in incubator, each administration group and the every Kong Jun of control group are separately added into
20 μ LMTT (5g/L), continue to be placed on 37 DEG C, 5%CO24h to be cultivated in incubator, abandons supernatant, 200 μ L DMSO are added in every hole,
Shaking table vibrates 10min at 37 DEG C, and microplate reader measures absorbance (A) value at 490nm.
6, mtt assay measures CIK to HepG2 killing functions of immunocytes
Using the CIK cell of culture to the 13rd day as effector cell, killing experiments in vitro is carried out.It is dense to adjust HepG2 cell
Degree 5 × 103A/mL, every 100 μ L of hole are divided into 1 enhanced sensitivity group of CM, 2 enhanced sensitivity group of CM, 3 enhanced sensitivity group of CM, 4 enhanced sensitivity group of CM, CM 5
Enhanced sensitivity group, 6 enhanced sensitivity group of CM and conventional group, CM 1-6 enhanced sensitivity group add 2 μM of compound CM 1-6 respectively, and conventional group is normal to cultivate,
It is placed on 37 DEG C, 5%CO2In incubator culture to cell it is adherent after change liquid, by effect target ratio 10:1 by effector cell be added cultivate
Kong Zhong, while it is right as feminine gender separately to set independent target cell (target cell+culture solution) and individual effect cell (effector cell+culture solution)
According to being placed on 37 DEG C, 5%CO2After acting on 48h in incubator, be added MTT (5g/L), 20 holes μ L/, continue to be placed on 37 DEG C,
5%CO24h is cultivated in incubator, 2000r/min is centrifuged 5min, and turnover panel abandons supernatant, and 150 hole μ L/ DMSO, concussion, to heavy is added
It forms sediment and dissolves, be put in microplate reader, killing activity is calculated as follows in 490nm densitometric (OD) value: killing rate (%)=
{ 1- [(experimental group OD value-individual effect cell OD value)/independent target cell OD value] } × 100%.
7, statistical method
It is analyzed using 19.0 statistical software of SPSS, data are indicated with mean ± standard deviation, and P < 0.05 anticipates with statistics
Justice.
Three, experimental result
1, CIK cell is proliferated
CIK cell is accelerated in the 5th day cell proliferation rate of culture, and the 5-13 days were the fast breeding period, at the 20th day
Cell Proliferation reaches 118 times of former plantation amount, reaches 176 times of former plantation amount in the 20th day cell Proliferation.
2, cytotoxicity of the compound CM 1-6 to HepG2 cell
Compound CM 1-6 administration group and absorbance (A) value of control group are without significant difference, it was demonstrated that 2 μM of compound CM 1-6
To HepG2 cell without obvious cytotoxicity.It the results are shown in Table 1.
The ratio of 1 compound CM 1-6 administration group of table and the absorbance value of control group
| CM 1 | CM 2 | CM 3 | CM 4 | CM 5 | CM 6 | |
| A administration/A control | 1.02 | 0.98 | 0.98 | 0.99 | 1.01 | 1.01 |
3, compound CM 1-6 enhances liver cancer cells to CIK cell killing sensibility
It can be seen that each enhanced sensitivity group CIK cell from table 2 and Fig. 1 and routine be all remarkably higher than to the killing rate of HepG2 cell
Group.Known 2 μM of compound CM 1-6 are to HepG2 cell without obvious cytotoxicity, and each enhanced sensitivity group CIK cell is to HepG2 cell
The raising of killing rate can only be attributed to the killing sensibility that compound CM 1-6 improves HepG2 cell to CIK cell, use change
After object CM 1-6 is closed to target cell HepG2 cell enhanced sensitivity, effect target is than in the case where constant, killing of the CIK cell to HepG2 cell
Power significantly improves.
Killing rate (effect target ratio 10:1) of the 2 each group CIK cell of table to HepG2 cell
Present invention demonstrates that pyridine-imidazole class compound, which can effectively enhance liver cancer cells, kills sensibility to CIK cell,
It can be prepared into pharmaceutically acceptable pharmaceutical preparation, just play preferably immune control using a small amount of CIK cell to realize
Therapeutic effect.
Above-described embodiment is the embodiment to essentiality content of the present invention, for preferably explaining the present invention, but this field skill
Protection scope of the present invention it is to be understood that above-mentioned specific embodiment should not be confined to by art personnel.
Claims (2)
1. a kind of pyridine-imidazole class organic compound is in drug of the preparation enhancing liver cancer cells to CIK cell killing sensibility
Application, which is characterized in that the pyridine-imidazole class organic compound have following general formula structure:
Wherein, R2For-OCH2CH3Or-OH；
R1For-CH2Or-CH (CH3)2Or-CH2CH2CH2-。
2. application according to claim 1, which is characterized in that pyridine-imidazole class organic compound is selected from following chemical combination
Object:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710290463.XA CN106943400B (en) | 2017-04-28 | 2017-04-28 | Enhance liver cancer cells to the compound and its preparation of CIK cell killing sensibility |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710290463.XA CN106943400B (en) | 2017-04-28 | 2017-04-28 | Enhance liver cancer cells to the compound and its preparation of CIK cell killing sensibility |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106943400A CN106943400A (en) | 2017-07-14 |
| CN106943400B true CN106943400B (en) | 2019-09-24 |
Family
ID=59477774
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710290463.XA Active CN106943400B (en) | 2017-04-28 | 2017-04-28 | Enhance liver cancer cells to the compound and its preparation of CIK cell killing sensibility |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106943400B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103626769A (en) * | 2013-12-23 | 2014-03-12 | 山东大学 | Substituted sulfydryl hexahydric heteroaromatic imidazole derivative and preparation method and application thereof |
| CN106083847A (en) * | 2016-08-03 | 2016-11-09 | 山东大学 | A kind of imidazopyridine mercapto phenylacetic acid derivative and preparation method and application |
-
2017
- 2017-04-28 CN CN201710290463.XA patent/CN106943400B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103626769A (en) * | 2013-12-23 | 2014-03-12 | 山东大学 | Substituted sulfydryl hexahydric heteroaromatic imidazole derivative and preparation method and application thereof |
| CN106083847A (en) * | 2016-08-03 | 2016-11-09 | 山东大学 | A kind of imidazopyridine mercapto phenylacetic acid derivative and preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106943400A (en) | 2017-07-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109498651A (en) | A kind of preparation method, medicament and the application of antitumor microparticle | |
| Andón et al. | Innate and adaptive responses of intratumoral immunotherapy with endosomal toll-like receptor agonists | |
| CN114870009A (en) | Anti-tumor combined composition, application thereof and anti-tumor medicine | |
| CN113599527B (en) | Application of APOE inhibitor and PD-1 monoclonal antibody in preparation of medicine for treating digestive tract tumor | |
| CN106943400B (en) | Enhance liver cancer cells to the compound and its preparation of CIK cell killing sensibility | |
| CN102688489B (en) | Pharmaceutical composition containing triptolide, triptolide derivative and Bc1-2 inhibitor and application thereof | |
| CN106974908A (en) | Pharmaceutical composition and purposes containing hdac inhibitor and IRE1 inhibitor | |
| CN109420167B (en) | Combined medicine for treating tumor | |
| CN102068448A (en) | Application of icariside II in preparation of anti-melanoma medicament | |
| Wang et al. | Early efficacy of stereotactic body radiation therapy combined with adoptive immunotherapy for advanced malignancies | |
| CN103284983A (en) | Application of alkannin and/or derivative thereof in preparing medicine for prohibiting cancer cell metastasis | |
| CN109528731A (en) | Pharmaceutical composition and its application with co-action for treating Huppert's disease | |
| CN113663081A (en) | Application of oroxylin and PD-1/PD-L1 inhibitor in preparation of liver cancer treatment drug | |
| Cao et al. | A preliminary study on the anticancer efficacy of Caulis spatholobi compound 1802. | |
| CN103599111B (en) | Combined drug for treating pancreatic cancer | |
| CN112603920A (en) | Application of traditional Chinese medicine toosendanin in preparation of products for preventing and treating glioma of nervous system | |
| CN102232957B (en) | Use of 3-acetoxyl-8, 24-lanostadiene-21-acid in preparing medicines for preventing or treating liver cancer or breast cancer | |
| CN106075454A (en) | A kind of anti-tumor medicinal preparation combination | |
| CN102688228A (en) | Pharmaceutical composition containing apigenin, apigenin derivative, rubescensin and rubescensin derivative, and application thereof | |
| CN106119192B (en) | Composition and its application in CIK cell culture | |
| Bocharova et al. | Prospects of using phytoadaptogen in the treatment of diffuse stomach cancer | |
| CN109432116A (en) | Astragaloside III is preparing the purposes in immunotherapy of tumors drug | |
| CN110856717A (en) | Tumor immunomodulator and application thereof | |
| CN108309994A (en) | Application of the magnesium isoglycyrrhetate in preparing antineoplastic immune regulation and control medicine | |
| CN110772513B (en) | Medicine for treating leukemia |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20190828 Address after: 050000 First Floor of Zhitong Pharmaceutical Valley Science Park, 197 Taihang South Street, Shijiazhuang High-tech Zone, Hebei Province Applicant after: Hebei Wanma Biomedical Co., Ltd. Address before: 211198 No. 18 Zhilan Road, Science Park, Jiangning District, Nanjing City, Jiangsu Province Applicant before: Nanjing cover Seef Pharmaceutical Technology Co. Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |