Dexamethasone is used to inhibit or treat the purposes of cancer of pancreas
Technical field
The present invention relates to the purposes of dexamethasone or its pharmaceutical salt or its preparation in treatment cancer of pancreas.
Background technique
One of the main reason for cancer (cancer) is whole world morbidity and is dead, according to the World Health Organization (World
Health Organization, WHO) recent statistics, there are about 8,800,000 cancer related mortalities, account for global death toll within 2015
16% (with reference to from WHO official website http://www.who.int/features/factfiles/cancer/en/).Its
In, cancer of pancreas is a kind of tumor in digestive tract for being difficult to diagnosing and treating, has the characteristics that grade malignancy height, poor prognosis, Huan Zhe
Cancer of pancreas advanced stage is often in when clinical definite, survival rate is only less than 3% within 5 years.
It mainly include operative treatment, radiotherapy, conventional cell poison at present for the therapeutic strategy of cancer of pancreas than relatively limited
Chemotherapy, the treatment of small molecule targeted drug etc., but curative effect is extremely limited.Patient only less than 20% is suitble to receive hand
Art treatment, and many patients after surgery can be dead due to recurrence;Pancreas cancer patients for classic chemotherapy drug response rate not yet
To 10%.
Dexamethasone (dexamethasone, DEX) is a kind of long-acting, synthetic glucocorticoid, has anti-inflammatory and immune suppression
The effects of processed, indication include a variety of inflammatories and autoimmune disease, allergic reaction, soft tissue oedema and shock rescue
Deng.In current existing research, any evidence there is no to show that DEX has good inhibiting effect to cancer of pancreas.
Summary of the invention
The present inventor has found DEX or its pharmaceutical salt or its preparation to cancer of pancreas in the therapeutic process of research cancer of pancreas
Growth there is good inhibitory effect, and without obvious drug toxicity.
DEX anti-inflammatory and immunosupress efficiency and preferable tolerance with higher, be widely used in treating inflammatory and
Autoimmune disease, allergic reaction and soft tissue oedema etc..In addition, DEX can be sent out for hematological cancer such as leukaemia etc.
Chemotherapy is waved, and it is mainly used for adverse reaction such as nausea and vomiting caused by alleviating chemicotherapy etc. in the treatment of solid tumor.
However, the research for influencing tumour progression on it all the time is less, the effect especially in cancer of pancreas is still unknown.The present invention
People has first carried out cell assay in vitro according to the classical thinking that anticarcinogen is studied at research initial stage, and DEX can be compared with as the result is shown
Inhibit the Clone formation of pancreatic cancer cell in vitro well (see embodiment 1).During testing in animal body, have studied respectively
DEX is antitumor in the animal model of transplantable tumor animal model and source of people tumor inoculation that pancreatic carcinoma PANC-1 is inoculated with
Effect has now surprisingly been found that DEX has significant inhibitory effect to cancer of pancreas, and effect is even better than clinical treatment of pancreatic cancer
In common chemotherapeutics gemcitabine (gemcitabine, GEM), and without obvious drug toxicity (see embodiment 2,3).
These results suggest that DEX shows good inhibiting effect and safety to the growth of cancer of pancreas.
Therefore, it can significantly inhibit cancer of pancreas it is an object of the present invention to provide a kind of and do not generate obvious toxic-side effects
Therapeutic agent, including a effective amount of dexamethasone or its pharmaceutical salt and preparation.
The pharmaceutical salt is to be synthesized by conventional chemical processes from parent compound, the parent compound packet
Containing alkalinity or acid part.In general, the salt is such as to be worked as by the free acid of these compounds or alkali form and chemistry
The suitable alkali or acid of amount in organic solvent or are reacted in water or in the mixture of water and organic solvent and are prepared.It is logical
Often, non-aqueous media such as ether, ethyl acetate, ethyl alcohol, isopropanol or acetonitrile is preferred.The example of acid-addition salts includes inorganic acid
Addition salts such as hydrochloride, hydrobromate, hydriodate, sulfate, nitrate, phosphate and organic acid addition salt such as acetate,
It is trifluoroacetate, maleate, fumarate, citrate, oxalates, succinate, tartrate, malate, flat
Peach hydrochlorate, analgin and tosilate.The example of base addition salts includes inorganic salts such as sodium, potassium, calcium and ammonium salt and organic
Alkali metal salt such as ethylenediamine, ethanol amine, N, the amino-acid salt of bis- alkylene ethanol amine of N-, triethanolamine and alkalinity.
According to an aspect of the present invention, the cancer of pancreas includes pancreatic sites tumour, tumour is formed and any other evil
Property tissue, preferably pancreas duct carcinoma.
It is a further object of the present invention to provide a kind of anticancer agents comprising the dexamethasone or its pharmaceutical salt
Customary adjuvant pharmaceutically.Wherein, dexamethasone conventional formulation includes tablet, solution, suspension, emulsion, powder, particle
Agent, capsule, micro-capsule, microballoon, injection, liposome, nanoparticle, slow/controlled release preparation etc..The customary adjuvant pharmaceutically
Including lubricant, filler, surfactant, solubilizer, cosolvent etc..
Realization of the invention has following positive effect:
(1) cancer of pancreas to be treated by DEX, tumour growth can be significantly inhibited, effect is significantly better than Common Chemotherapy drug, and
Without obvious toxic-side effects;
(2) drug safety of the present invention is good, and patient can be greatly reduced under conditions of ensuring good drug effect
Operative risk.
Detailed description of the invention
Fig. 1 is influence result figure of the DEX to human pancreatic carcinoma PANC-1 cell line clonality.PANC-1 cell is given respectively
The photo of cell clonal formation when giving blank control and various concentration DEX.
Fig. 2 is influence result figure of the DEX to human pancreatic carcinoma PANC-1 cell line clonality.PANC-1 cell is given respectively
The quantitative result of cell clonal formation when giving blank control and various concentration DEX (with blank control group for 100%).
Fig. 3 is the growth curve (DEX multi-dose, n=5) of female lotus knurl NOD/SCID mouse tumor when DEX is administered.Source of people
Pancreatic Adenocarcinoma inoculation tumor-bearing mice give respectively blank control, DEX 0.5mg/kg, DEX 2mg/kg, DEX 4mg/kg,
The change curve of mouse tumor volume when gemcitabine (GEM) 15mg/kg.
Fig. 4 is the photo (DEX multi-dose, n=5) of female lotus knurl NOD/SCID mouse tumor when DEX is administered.Source of people pancreas
The tumor-bearing mice of cancerous tissue inoculation gives blank control, DEX 0.5mg/kg, DEX 2mg/kg, DEX 4mg/kg, Ji Xi respectively
The 34th day tumour real shooting photo of mouse tumor when his shore (GEM) 15mg/kg.
Fig. 5 is the changes of weight (DEX multi-dose, n=5) of female lotus knurl NOD/SCID mouse when DEX is administered.Source of people pancreas
The tumor-bearing mice of cancerous tissue inoculation gives blank control, DEX 0.5mg/kg, DEX 2mg/kg, DEX 4mg/kg, Ji Xi respectively
The change curve of mouse weight when his shore (GEM) 15mg/kg.
Fig. 6 is the organ index (DEX multi-dose, n=5) of female lotus knurl NOD/SCID mouse when DEX is administered.Source of people pancreas
The tumor-bearing mice of cancerous tissue inoculation gives blank control, DEX 0.5mg/kg, DEX 2mg/kg, DEX 4mg/kg, Ji Xi respectively
His the 34th day percentage of heart, liver, spleen, lungs, kidney weight relative to mouse weight of shore (GEM) 15mg/kg.
Fig. 7 is the blood picture index (DEX multi-dose, n=5) of female lotus knurl NOD/SCID mouse when DEX is administered.Source of people pancreas
The tumor-bearing mice of cancerous tissue inoculation gives blank control, DEX 0.5mg/kg, DEX 2mg/kg, DEX 4mg/kg, Ji Xi respectively
He shore (GEM) 15mg/kg the 34th day when mouse blood picture index.
Fig. 8 is the growth curve (DEX single dose, n=4) of female tumor bearing nude mice tumour when DEX is administered.Human pancreas cancer
The change curve of nude mouse tumor volume when PANC-1 tumor bearing nude mice gives blank control or DEX 2mg/kg respectively.
Fig. 9 is the photo (DEX single dose, n=4) of female tumor bearing nude mice tumour when DEX is administered.Human pancreas cancer PANC-1 lotus
Tumor nude mice gives blank control or DEX 2mg/kg the 26th day tumour real shooting photo respectively.
Figure 10 is the variation (DEX single dose, n=4) of female tumor bearing nude mice weight when DEX is administered.Human pancreas cancer PANC-1
The change curve of nude mice weight when tumor bearing nude mice gives blank control or DEX 2mg/kg respectively.
Figure 11 is the organ index (DEX single dose, n=4) of female tumor bearing nude mice when DEX is administered.Human pancreas cancer PANC-1
Tumor bearing nude mice give respectively the 26th day heart of blank control or DEX 2mg/kg, liver, spleen, lungs, kidney weight relative to
The percentage of nude mice weight.
Specific embodiment
In order to better understand the present invention, current inventor provides following embodiments, however, these embodiments are only
Illustratively purpose and provide, and be not interpreted as limitation of the present invention because many variations be it is possible,
Without departing from the spirit and scope of the invention.Although method of the invention is described in specific embodiment, affiliated technology
The technical staff in field is it is clear that the step of can make the method or the sequence of step changes, but without departing from the present invention
Concept and scope.More specifically, it is clear that it is similar with other biologically relevant pharmacology processes in chemistry
Drug can replace drug as described herein, while reach the same or similar effect.The technical personnel in the technical field are aobvious
And what is be clear to should all be considered in scope of the invention and concept all this similar replacements or modification.
Embodiment 1:
The effect that DEX forms human pancreatic cancer cell body outer clone
The present embodiment has studied influence of the DEX to human pancreatic cancer cell clonality in vitro.
The PANC-1 pancreatic cancer cell that form is normal, in good condition is taken, the operation digested, be centrifuged, is suspended again is simultaneously
It counts.According to count results, cell suspension is diluted to 1000/mL with culture medium, 6 holes are added to the volume of every hole 2mL
In tissue culture plate, make 2000, every hole cell.6 orifice plates after inoculation are placed in cell incubator.
DEX titer is diluted with culture medium respectively, respectively obtains that DEX is 1 μM final concentration of, 10 μM, the training of 100 μM of drug containing
It is spare to support base.Isometric DMSO is added in the culture medium of blank control group, and makes the DMSO concentration in system not higher than 1%.
It is administered after 72h, inhales and abandon each hole culture medium, every hole is added 2mL fresh culture, changes the liquid once within every 2 days.Inoculation 10
It after it, inhales and abandons culture medium, 0.5mL PBS rinsing is added once in every hole, inhales every hole after abandoning PBS and 0.5mL methanol is added, solid at room temperature
Determine 10min.It inhales and abandons methanol, dye 10min with 0.5% crystal violet solution.It inhales and abandons crystal violet solution, rinse 6 orifice plates with tap water
To wash away unbonded dyestuff, naturally dry.6 orifice plates after drying are taken pictures, gram of ImageQuant TL7.0 software is utilized
Grand tally function counts the clone's quantity to be formed, and is compared.
(Fig. 1,2) as the result is shown:DEX has the clonality of human pancreatic cancer cell PANC-1 in vitro more significant
Influence;DEX concentration is higher, and the effect for inhibiting tumor cell clone to be formed is stronger.
It discusses:Colony formation mainly investigates the ability that individual cells are proliferated in vitro and form colony, indirectly
The Tumor formation of tumour cell is reacted;Drug by the way that various dose is added in the medium can investigate cell in drug effect
The lower ability for maintaining fertility, has had researcher to grind the tumor stem cell that the method is used for including cancer of pancreas at present
Study carefully.This chapter research in colony formation the result shows that, DEX has significantly under 1 μM, 10 μM and 100 μM of concentration
Inhibit clonality, there is dose dependent, can illustrate that DEX has the function of inhibiting tumor development.
Embodiment 2:
Inhibiting effect (DEX multi-dose) of the DEX to tumour growth in internal cancer of pancreas source of people Transplanted tumor model
The present embodiment has studied the inhibiting effect for the mice tumors grew that DEX is inoculated with clinical patients tumor tissues.
By the obtained Pancreatic Adenocarcinoma of excision of performing the operation out of clinical patients body, take out a part be cut into about 2mm ×
The fritter of 2mm × 3mm is injected into the NOD/SCID right side of mice subcutaneous abdomen of about 6 week old with the medullo-puncture needle of sterilizing, and referred to as
Generation source of people xenograft tumor (patient-derived xenograft, PDX) model mouse (F1);To tumour growth to about
500mm3When, it puts to death mouse and takes out tumour rapidly, a part is placed in cryopreservation tube, the FBS containing 10%DMSO is added, is placed in liquid
It is frozen in nitrogen;Another part tumour is cut into 2mm × 2mm × 3mm fritter, is seeded to NOD/SCID mouse skin according to the method described above
Under, referred to as second generation PDX model mouse (F2), and so on.This experiment PDX model mouse used is the third generation (F3).
After third generation inoculation, observation inoculation situation.With the major diameter (D of vernier caliper measurement tumourmax) and minor axis (Dmin), meter
Calculate gross tumor volume V (V=Dmax×Dmin 2/2).When tumour growth to 100~200mm3When mouse is randomly divided into 5 groups, every group 5
Only.Every group of dosage regimen is respectively:(1) blank control group:Daily 10% hydroxypropyl-β-cyclodextrin solution of stomach-filling 0.1mL;
(2) GEM group:Every physiological saline of the tail vein injection 0.1mL on the three containing GEM, GEM dosage are 15mg/kg;(3)DEX
0.5mg/kg group:Daily 10% hydroxypropyl-β-cyclodextrin solution of the stomach-filling 0.1mL containing DEX, the dosage of DEX are 0.5mg/
kg;(4) DEX 2mg/kg group:Daily 10% hydroxypropyl-β-cyclodextrin solution of the stomach-filling 0.1mL containing DEX, the dosage of DEX
For 2mg/kg;(5) DEX 4mg/kg group:Daily 10% hydroxypropyl-β-cyclodextrin solution of the stomach-filling 0.1mL containing DEX, DEX's gives
Pharmaceutical quantities are 4mg/kg.Wherein, blank control group is negative control group, and GEM (gemcitabine) is positive controls.
Since the 0th day, the survival condition of every group of animal is observed, if obvious adverse reaction occur, and equaled a record with day
Its weight is administered daily the foundation of dosage as calculating, and quantifies to reflect the survival condition of animal.It is complete the 34th day after administration
After measurement, blood routine measurement is carried out to control group and test group of animals blood sampling, takes the method for blood from the mouse right side using eye socket
Eye takes out 20 μ L of whole blood, is rapidly added in 5mL dilution, slight oscillatory upper machine measurement after mixing.Animal is put to death, tumour is driven
It is taken out in object, puts and take pictures by group.After putting to death animal, completely its heart, liver, spleen, lungs, kidney are taken out in dissection
It is dirty, it is cleaned up with physiological saline, and weigh after being dried with filter paper, calculates the organ index of each internal organs of animal, calculation formula is:
(Fig. 3,4,5,6,7) as the result is shown:DEX each group gross tumor volume is significantly less than blank control group, and each dosage group of DEX
Drug effect be superior to GEM positive controls;Dose dependent is presented in the antitumor drug effect of DEX.Mouse weight is overall during administration,
There was no significant difference with blank control group weight, and main organs are without obvious damage, blood picture indices Non Apparent Abnormality.The above knot
Fruit shows that DEX can show preferable tumor inhibition effect in the xenograft tumor that source of people tumor tissues are inoculated with, and drug is pacified
Full property is good.
It discusses:Source of people xenograft tumor (PDX) model is a kind of model common in preclinical study.PDX model with
Traditional nude mouse xenograft tumor model is compared, and main advantage is to be inoculated with tumor mass from clinical patients, therefore its tumour
The characteristics of growth, is closer with human tumor.Some researches show that until F7~F10, PDX Model Tumor still can be with Primary Tumor
Keep higher similitude.In the PDX animal model that the present embodiment is established, DEX has shown preferable antitumor drug effect,
Meanwhile during administration weight without significant change, show DEX without obvious general toxicity, organ index the result shows that DEX to main
Internal organs are without significant toxicity, and the indices and control group of DEX each group there are no significant difference, show its safety in blood picture result
Preferably.
Embodiment 3:
Inhibiting effect (DEX single dose) of the DEX to tumour growth in internal cancer of pancreas Transplanted tumor model
The present embodiment has studied the inhibiting effect for the nude mouse tumor growth that DEX is inoculated with human pancreatic cancer cell.
The good PANC-1 cell of growth conditions is taken, the DMEM culture medium without FBS on a small quantity is added after digestion, with elbow glass
Glass suction pipe blows down the cell being attached in culture bottle wall, is transferred to 15mL centrifuge tube, and 1000rpm is centrifuged 5min, discards supernatant,
It a small amount of culture medium is added blows and beats cell again and mix and count, according to cell counts by the cell suspension training of no FBS
Feeding base is diluted to 5 × 107A/mL.Diluted cell suspension is injected to oxter on the right side of nude mice according to the dimensions subcutaneous of 0.2mL
(about contain 1 × 107A cell), observation inoculation situation.
After inoculation, tumour growth to about 100mm3When mouse is randomly divided into 2 groups, every group 4.Every group of dosage regimen
For:(1) blank control group:Daily 10% hydroxypropyl-β-cyclodextrin solution of stomach-filling 0.1mL;(2) DEX 2mg/kg group:It is daily to fill
10% hydroxypropyl-β-cyclodextrin solution of the stomach 0.1mL containing DEX, the dosage of DEX are 2mg/kg.Wherein, blank control group is
Negative control group.
Since the 0th day, the survival condition of every group of animal is observed, if obvious adverse reaction occur, and equaled a record with day
Its weight is administered daily the foundation of dosage as calculating, and quantifies to reflect the survival condition of animal.It is complete the 26th day after administration
After measurement, animal is put to death, tumour is taken out out of animal body, puts and takes pictures by group.After putting to death animal, completely dissect
Its heart, liver, spleen, lungs, kidney are taken out, is cleaned up with physiological saline, and is weighed after being dried with filter paper, animal is calculated
The organ index of each internal organs, calculation formula are shown in formula 1.
(Fig. 8,9,10,11) as the result is shown:DEX 2mg/kg group gross tumor volume is significantly less than blank control group, DEX administration
There was no significant difference with blank control group for period nude mice weight, and main organs are also without obvious damage.The above result shows that DEX is in people
Preferable tumor inhibition effect can be showed in the xenograft tumor of pancreatic cancer cell inoculation, and drug safety is good.
It discusses:In the human pancreas cancer xenograft tumor of PANC-1 cell inoculation, DEX has shown preferable antineoplastic
Effect.DEX shows that its safety is preferable without obvious whole body or organ toxicity simultaneously.