CN108992672A - A kind of wogonin pharmaceutical composition for treating neoplastic disease - Google Patents
A kind of wogonin pharmaceutical composition for treating neoplastic disease Download PDFInfo
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Abstract
The present invention relates to a kind of wogonin pharmaceutical composition for treating neoplastic disease, described pharmaceutical composition includes that the component of following parts by weight proportion is made: 20~30 parts of the wogonin, 80~120 parts of L-arginine, 30~50 parts of mannitol, 2200~2800 parts of water for injection；Pharmaceutical composition of the present invention has significant in efficacy, the small advantage of toxic side effect.
Description
Technical field
The invention belongs to field of pharmaceutical preparations more particularly to a kind of wogonin pharmaceutical compositions for treating neoplastic disease.
Background technique
Malignant tumour is a kind of disease for seriously threatening human health, and disease incidence rises year by year in recent years, in many prosperities
Country, mortality of malignant tumors are only second to heart disease, column Proportional Death Rate forefront.In China, malignant tumour has also been included in first and second
The cause of the death.Malignant tumour seriously endangers human health, though complex treatment can be carried out by operation, chemotherapy, radiotherapy etc., cure rate is still
It is very low.And the selectivity of anticancer Western medicine that clinic is often used is not high at present, while killing tumor cell just to body
The cell of normal cell, especially molecular marker for increased proliferation has detrimental effect.The long-time service of these drugs, also brought to tumor patient compared with
Big toxic side effect causes this kind of drug compliance bad, thus seeks to develop a kind of drug of the treatment tumour of high-efficiency low-toxicity
It is very necessary.
Wogonin is one of the effective component of Chinese traditional herbs radix scutellariae as a kind of natural flavone compound, tool
There are a variety of effects such as antibacterial, antiviral and antitumor, antipyretic-antalgic, anti-oxidant and scavenging activated oxygen and treatment cardiovascular disease.
The a variety of cultured tumor cells in vitro of wogonin (human stomach cancer cell line MGC-803 and BGC-823, HepG2 cell lines,
Human liver cancer 5-FU persister BEL-7402/5-FU, human colon cancer cell strain SW1116 and HCT116, human prostate cancer cell line
DU145, human lung carcinoma cell line H460, Breast cancer lines MDA-MB-231 and MCF-7, people's malignant melanoma cell strain
A375, human leukemia cell line HL60 and K562 and hereditary variation cell strain NIH Swiss mouse blastocysts NIH-3T3)
Growth has apparent inhibiting effect, but influences on normal cell smaller.Since content of the wogonin in baikal skullcap root is lower.
Using inexpensive pyrogallic acid as Material synthesis wogonin, (specific synthesis technology can be found in the special of CN1915987A for we
Benefit), and be large-scale production, raw material is provided.But there are the dosage forms such as tablet, capsule, injection for facing for wogonin at present
Bed uses, these dosage forms are poor in gastrointestinal tract dissolution rate, and bioavilability is very low.Wogonin is unstable in the solution, and one
Injection can not directly be made, therefore exploitation is badly in need of one kind and has good stability, the small preparation of toxic side effect.
Summary of the invention
The purpose of the present invention is to provide a kind of wogonin pharmaceutical composition for treating neoplastic disease, pharmaceutical composition tools
Have the advantages that quality is reliable and stable, significant in efficacy, toxic side effect is small.
Wogonin raw material used in the present invention is made by chemically synthesized method, and specific method is coke galla turcica
Acid process is Benzylation, reoxidizes, restores, methylating, and is prepared into intermediate, then hydrogenolysis sloughs the series of steps such as benzyl, obtains
To the wogonin raw material of purity > 99%.
The technical solution of present patent application are as follows:
Pharmaceutical composition of the present invention includes the component of following parts by weight proportion: 10~40 parts of wogonin, hydrotropy
20~200 parts of agent, 20~60 parts of framework material, 2000~3000 parts of water for injection.
Preferably, the parts by weight proportion that described pharmaceutical composition is are as follows: 20~30 parts of wogonin, cosolvent 80~
120 parts, 30~50 parts of framework material, 2200~2800 parts of water for injection.
Cosolvent is one of sodium bicarbonate, glycerol, sodium phosphate, L-arginine, citric acid in described pharmaceutical composition
Or it is a variety of.
Framework material in described pharmaceutical composition is one of lactose, glucose, sorbierite, mannitol or a variety of.
Described pharmaceutical composition includes the component of following parts by weight proportion: 25 parts of the wogonin, L-arginine 100
Part, 40 parts of mannitol, 2500 parts of water for injection.
The dosage form of described pharmaceutical composition is freeze-dried powder or injection.
The preparation method of pharmaceutical composition of the present invention, comprises the following steps that:
(1) after, mixing wogonin and cosolvent plus a small amount of water for injection moistens, and water for injection stirring, which is added, makes drug
All dissolutions, obtain mixed solution；
(2), take the mixed solution of step (1) to add framework material to be stirred to dissolve, active carbon is added and stirs depyrogenation, takes off
Charcoal filters, and injection appropriate amount of water constant volume is added to configure；Again through membrane filtration degerming, filtering liquid medicine is obtained, it is freeze-dried at freeze-dried powder,
?.
The preparation method processing step (2) in freeze drying process condition are as follows: in cryogenic temperature be -60~-40 DEG C it is cold
Freeze 3~6h of time；And cryogenic temperature -30~-0 DEG C is maintained, 10~20pa of vacuum degree, cooling time is 15~30h；Continue to keep
10~20pa of vacuum degree, 0~20 DEG C of cryogenic temperature, cooling time is 4~8h；Then 25~35 DEG C, when dry are gradually heated to
Between 5~8h, vacuum tamponade can obtain freeze-dried powder.
The preparation method technique carries out in a nitrogen environment.
Present inventor after a lot of trial and error, finds quality finally and stablizes, the preferable wogonin of curative effect
Pharmaceutical composition prescription is that the present invention tests preferred part research data below.
The screening of 1.1 cosolvents
The wogonin and cosolvent for taking recipe quantity are set in container, and appropriate water for injection is added, and heating makes to dissolve, and add note
It penetrates with water to enough.
The screening of 1 cosolvent of table
As a result it and analyzes: using a variety of different cosolvents during the test, and adjust dosage, as a result carbon therein
Sour hydrogen sodium and arginine can be such that wogonin all dissolves.But when using sodium bicarbonate, drug solution is when being placed at room temperature for one section
Between after, crystallization is precipitated in drug quickly, illustrates that its solubilization-aid effect is bad；And use arginic solubilization-aid effect good, it can must clarify
The transparent and stable drug solution not being precipitated under room temperature, but when arginine amount is excessive, causes the pH value of solution excessively high, it may
Be not suitable for clinical use；When wogonin and arginic weight ratio are 1:4, drug can be completely dissolved, and the pH of medical fluid
Value is suitable for, it is thus determined that cosolvent is arginine in prescription, dosage is every bottle of 100mg.
The screening of 1.2 framework materials
It takes the wogonin 25mg and L-arginine 100mg of recipe quantity to set in container, the water for injection boiled is added to stir, and
100 DEG C are maintained to its dissolution；Appropriate mannitol is added after letting cool to be stirred to dissolve, benefit injects water to water for injection extremely
2.5mL enough.Measure the osmotic pressure of each medical fluid；Each medical fluid is sub-packed in cillin bottle, send and is freeze-dried into freeze dryer, is investigated
The mouldability and solubility of wogonin.
The content of mannitol and the relationship of osmotic pressure, mouldability and solubility in 2 wogonin solution of table
Illustrate: the inspection method of solubility is the addition 2.5mL distilled water in freeze-dried powder, is then shaken, and freeze-dried powder is observed
Dissolution situation.As a result, can all be dissolved using the freeze-dried powder of above-mentioned each prescription preparation, but solution rate is different.I
Indicate that dissolution is slow with *；* indicates that dissolution is slower；* * indicates that dissolution is very fast；* * * indicates quickly dissolution.
Above-mentioned experimental result table 2 shows to be incremented by with mannitol dosage, and osmotic pressure is gradually increased.It is in mannitol dosage
When 40-60mg, the osmotic pressure of solution meets infiltration pressure request between 280-320mOsm/kg, it is contemplated that mouldability, solubility
Etc. indexs, the final dosage for determining mannitol in prescription be that 40mg is added in every 2.5mL solution.
1.3 miillpore filter adsorption tests are investigated
It takes the wogonin 25mg and L-arginine 100mg of recipe quantity to set in appropriate vessel, the water for injection boiled is added
Make to dissolve to 2.5ml.Mannitol 40mg is added to stir evenly, sample solution is divided into two parts, portion is another without any processing
For part through 0.22 μm of filtering with microporous membrane, two parts of solution measure UV absorption after suitably diluting at 275nm, investigate micropore filter
Whether film filtering front and back drug has loss.It the results are shown in Table 3.
Result is investigated in 3 miillpore filter adsorption test of table
Before and after above-mentioned test result shows filtering with microporous membrane, drug is not lost, and illustrates 0.22 μm of miillpore filter to master
Medicine can be used for the aseptic filtration of this technique herb liquid without absorption.
1.4 charcoal tests are investigated
The drug solution prepared by above-mentioned prescription is divided into four parts, is added respectively by 0.0%, 0.1%, 0.3%, 0.5%
Enter needle-use activated carbon, stirring takes off charcoal after twenty minutes, and solution measures UV absorption after suitably diluting at 275nm, investigates activity
Absorption situation of the charcoal to drug.It the results are shown in Table 4.
4 charcoal test result of table
Above-mentioned test result shows to increase with concentration of activated carbon, also gradually increases to the adsorbance of drug, therefore removing
While pyrogen, the active carbon of low concentration should be selected as far as possible.It proceeds from reality, the active carbon of final choice 0.1%.Together
When to guarantee that the medicament contg in final products, the dosage of wogonin increase by 10%.
The drafting of 1.5 freeze-drying curves
It takes the wogonin 25mg and L-arginine 100mg of recipe quantity to set in appropriate vessel, the water for injection boiled is added
Make to dissolve to 2.5ml.Mannitol 40mg is added to stir evenly, after filtering with microporous membrane, obtains the solution of injection wogonin,
Sample to be lyophilized is set in freeze dryer, eutectic point and temp probe are inserted into, closes chamber door, freeze dryer is opened, utilizes conduction oil
Decline product temperature, the eutectic point of this product is surveyed, about at -11 DEG C or so.So the temperature of sample is brought down below -38 DEG C, heat preservation 2 is small
When, it is then turned on condenser, when condensation temperature is -40 DEG C hereinafter, opening vacuum system, vacuum control 15Pa ± 4Pa again；It will be thermally conductive
Oil temperature rises to -11 DEG C, keeps the temperature 14 hours, after products temperature rises to eutectic temperature 1 hour, distillation terminates.Distillation terminates
Thermally conductive oil temperature is risen to 40 DEG C afterwards, products temperature rises to 30 DEG C, and timing keeps the temperature 6 hours, tamponade, it deflates, outlet.
1.6 pilot scale sample preparations
On the basis of optimization, test agent in preparation three batches, formulation and technology is as follows: take wogonin 25mg with
L-arginine 100mg is set in appropriate vessel, and the water for injection 2200ml boiled is added, is stirred to dissolve；Mannitol 40mg is added
It is stirred to dissolve；Needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and filters into clean container, mend
Inject water to 2500ml, solution stirring made uniformly for 5 minutes, then through 0.22 μm of filtering with microporous membrane, filtrate it is filling
In 10ml cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send on the plate layer to freeze drying box, is inserted into temperature
Probe closes chamber door.It is lyophilized by freeze-drying curve, as a result the investigation of three batches of samples see the table below 5.
5 three batches of sample yield rates of table and appearance are investigated
1.7 Photostability experiments
It takes suitable wogonin and L-arginine to set in container in prescription ratio, the water for injection boiled is added to stir, and
100 DEG C are maintained to its dissolution, removes and lets cool to room temperature, mannitol is added, is stirred to dissolve, benefit injects water to enough.
When being denoted as zero；It places under the illumination of 4500Lx for 24 hours, samples at regular intervals, measure the content of sample, in relation to substance and pH
Value.Wogonin solution is investigated to the stability of light.It the results are shown in Table 6.
The photostability of 6 wogonin solution of table
The above results table 6 show strong illumination for 24 hours after, this product content does not have a significant change, and related substance slightly increases
Add, pH value of solution does not have significant change, illustrates that wogonin solution is more stable, the production process of wogonin preparation to light
Without being protected from light.
1.8 stability of the production process are investigated
Wogonin 25mg, L-arginine 100mg, mannitol 40mg are taken in prescription ratio, water for injection to 2.5mL takes
Suitable wogonin and L-arginine are set in container, add the water for injection boiled stir, and be maintained at 100 DEG C to its dissolution,
It removes and lets cool to room temperature, mannitol is added, is stirred to dissolve, benefit injects water to enough.By the wogonin of above-mentioned preparation
Solution is placed under the conditions of 25 DEG C, is sampled at regular intervals, measures the content of sample, in relation to substance and pH value.It the results are shown in Table 7.
The stability of 7 wogonin solution of table in process of production
Interpretation of result: groping by technique early period, primarily determined injection wogonin pharmaceutical composition prescription and
Production technology.In view of being formulated into filling and cartonning freezing from medical fluid in actual production process, it may be necessary to a period of time,
Therefore the stability of medical fluid in the process need to be investigated.As a result, this product solution is placed for 24 hours under the conditions of 25 DEG C, content is without bright
Aobvious variation, related substance meet bound requirements, and the pH value of solution does not have significant change, illustrates that this product production stability is preferable, work
Skill is feasible.
The application's has the beneficial effect that
On the basis of existing technology, present inventor has carried out wogonin injection prescription and has further ground
Study carefully screening, finishing screen selects a kind of quality stabilization, reliable wogonin pharmaceutical composition.Pharmaceutical composition prescription of the present invention is matched
Than reasonable, pharmacological experiment is proved: the application wogonin pharmaceutical composition is to human gastric cancer MGC-803 tumour cell, human lung cancer
H460 nude mouse xenograft tumor, liver cancer SMMC-7721 tumour cell, human gastric cancer BGC-823 nude mouse xenograft tumor, human liver cancer
The kinds of tumor cells such as SMMC-7721 nude mouse xenograft tumor have apparent growth inhibition effect, this also indicates that medicine of the present invention
Compositions are significant in efficacy.Toxicological experiment the result shows that, the small advantage of pharmaceutical composition toxic side effect of the present invention.
It has good stability according to wogonin injection preparation, reliable in quality made from currently preferred preparation method.
Specific embodiment
It is the specific embodiment of the content of present invention below, for illustrating the skill of the technical problem of being solved in present specification
Art scheme facilitates those skilled in the art understand that the content of present invention, but the realization of technical solution of the present invention is not limited to these
Embodiment.It should carried out in a nitrogen environment in the preparation process for implementing 1-9.
Embodiment 1
It takes 25g wogonin and 100g L-arginine is set in appropriate vessel, it is appropriate to be added the water for injection boiled, stirring
Make to dissolve；Mannitol 40g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core
It filters in clean container, benefit injects water to 2500ml, and solution stirring is made uniformly, then through 0.22 μm of micropore for 5 minutes
Membrane filtration, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send to freeze drying box
On plate layer, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -50 DEG C of cooling time 4h；And -20 DEG C of cryogenic temperature are maintained,
Vacuum degree 15pa, cooling time 15h；Continue keep vacuum degree 15pa, -10 DEG C of cryogenic temperature, cooling time 6h；Finally do
Dry temperature is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 2
It takes 30g wogonin and 120g L-arginine is set in appropriate vessel, it is appropriate to be added the water for injection boiled, stirring
Make to dissolve；Mannitol 50g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core
It filters in clean container, benefit injects water to 2200ml, and solution stirring is made uniformly, then through 0.22 μm of micropore for 5 minutes
Membrane filtration, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send to freeze drying box
On plate layer, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -60 DEG C of cooling time 3h；And 0 DEG C of cryogenic temperature is maintained, very
Reciprocal of duty cycle 20pa, cooling time 30h；Continue keep vacuum degree 10pa, 0 DEG C of cryogenic temperature, cooling time 4h；Finally dry temperature
Degree is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 3
It takes 20g wogonin and 80g L-arginine is set in appropriate vessel, be added that the water for injection boiled is appropriate, and stirring makes
Dissolution；Mannitol 30g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and takes out
It filters in clean container, benefit injects water to 2800ml, and solution stirring is made uniformly for 5 minutes, then filters through 0.22 μm of micropore
Film filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate to freeze drying box
On layer, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -40 DEG C of cooling time 6h；And -30 DEG C of cryogenic temperature are maintained, very
Reciprocal of duty cycle 10pa, cooling time 15h；Continue keep vacuum degree 20pa, -20 DEG C of cryogenic temperature, cooling time 8h；Finally dry
Temperature is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 4
It takes 40g wogonin and 200g L-arginine to set in appropriate vessel, the water for injection boiled is added, stirring makes molten
Solution；Mannitol 60g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and filters
Into clean container, benefit injects water to 3000ml, and solution stirring is made uniformly, then through 0.22 μm of miillpore filter for 5 minutes
Filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate layer to freeze drying box
On, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -50 DEG C of cooling time 5h；And -20 DEG C of cryogenic temperature are maintained, vacuum
Spend 15pa, cooling time 20h；Continue keep vacuum degree 15pa, -10 DEG C of cryogenic temperature, cooling time 5h；Finally dry temperature
Degree is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 5
It takes 10g wogonin and sodium phosphate 20g, set in appropriate vessel, be added that the water for injection boiled is appropriate, and stirring makes molten
Solution；Mannitol 20g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and filters
Into clean container, benefit injects water to 2000ml, and solution stirring is made uniformly, then through 0.22 μm of miillpore filter for 5 minutes
Filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate layer to freeze drying box
On, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -55 DEG C of 3~6h of cooling time；And -10 DEG C of cryogenic temperature are maintained,
Vacuum degree 15pa, cooling time 15h；Continue keep vacuum degree 20pa, -5 DEG C of cryogenic temperature, cooling time 6h；Finally do
Dry temperature is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 6
It takes 30g wogonin and citric acid 120g, set in appropriate vessel, be added that the water for injection boiled is appropriate, and stirring makes
Dissolution；Glucose 50g is added, is stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and takes out
It filters in clean container, benefit injects water to 2500ml, and solution stirring is made uniformly for 5 minutes, then filters through 0.22 μm of micropore
Film filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate to freeze drying box
On layer, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -45 DEG C of cooling time 4h；And -25 DEG C of cryogenic temperature are maintained, very
Reciprocal of duty cycle 20pa, cooling time 15h；Continue keep vacuum degree 10pa, -10 DEG C of cryogenic temperature, cooling time 7h；Finally dry
Temperature is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 7
It takes 20g wogonin and citric acid 80g is set in appropriate vessel, be added that the water for injection boiled is appropriate, and stirring makes molten
Solution；Glucose 30g is added, is stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and filters
Into clean container, benefit injects water to 2000ml, and solution stirring is made uniformly, then through 0.22 μm of miillpore filter for 5 minutes
Filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate layer to freeze drying box
On, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -60~-40 DEG C of 3~6h of cooling time；And maintain cryogenic temperature-
30~-0 DEG C, 10~20pa of vacuum degree, cooling time is 15~30h；Continue keep 10~20pa of vacuum degree, cryogenic temperature 0~-
20 DEG C, cooling time is 4~8h；Last drying temperature is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 8
It takes 40g wogonin and sodium bicarbonate 200g is set in appropriate vessel, be added that the water for injection boiled is appropriate, and stirring makes
Dissolution；Lactose 60g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and filters
Into clean container, benefit injects water to 2200ml, and solution stirring is made uniformly, then through 0.22 μm of miillpore filter for 5 minutes
Filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate layer to freeze drying box
On, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -50 DEG C of cooling time 5h；And -20 DEG C of cryogenic temperature are maintained, vacuum
Spend 15pa, cooling time 20h；Continue keep vacuum degree 15pa, -10 DEG C of cryogenic temperature, cooling time 7h；Finally dry temperature
Degree is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
Embodiment 9
It takes 10g wogonin and sodium bicarbonate 20g is set in appropriate vessel, be added that the water for injection boiled is appropriate, and stirring makes
Dissolution；Sorbierite 20g is added to be stirred to dissolve, needle-use activated carbon is added by 0.1%, stirs 20 minutes, takes off charcoal through titanium sand core and takes out
It filters in clean container, benefit injects water to 2500ml, and solution stirring is made uniformly for 5 minutes, then filters through 0.22 μm of micropore
Film filtering, filtrate is filling in cillin bottle, every bottle of 2.5ml, then partly butyl rubber bung beyond the Great Wall, send the plate to freeze drying box
On layer, it is inserted into temp probe, closes chamber door.In cryogenic temperature be -60 DEG C of cooling time 3h；And -30 DEG C of cryogenic temperature are maintained, very
Reciprocal of duty cycle 20pa, cooling time 15h；Continue keep vacuum degree 15pa, -20 DEG C of cryogenic temperature, cooling time 5h；Finally dry
Temperature is 30 DEG C and is kept for 6 hours.Close plug, deflates, and outlet rolls lid.
In order to further verify the currently preferred pharmacology and toxicity curative effect for implementing 1 group of wogonin pharmaceutical composition,
The related pharmacology and toxicological test testing data of following the application wogonin pharmaceutical composition.
2. pharmacological experiment
2.1 wogonin pharmaceutical compositions (1 group of the embodiment of the present invention) of the present invention are applied with chemotherapy drugs in combination to external
Cultivate the growth inhibition effect of human gastric cancer MGC-803 tumour cell
Investigation wogonin pharmaceutical composition and chemotherapeutics fluorouracil (5-FU), cis-platinum (DDP), adriamycin (EPB),
Growth inhibition effect of taxol (PTX) use in conjunction to vitro culture of human Gastric Cancer MGC -803.
The result shows that wogonin pharmaceutical composition and its with 5-FU by IC50 ratio (3:1) be administered in combination, wogonin
The drug combination in 22.25 μM+8.9 μM to 200 μM+80 μM of dosage range cooperates with work to pharmaceutical composition with fluorouracil
With.Wogonin pharmaceutical composition and its with cis-platinum by IC50 ratio (1:4) be administered in combination, wogonin pharmaceutical composition with
Drug combination has synergistic effect in the dosage range that 60 μM+45 μM to 2.42 μM+1.82 μM of end level of cis-platinum.Wogonin medicine
Compositions and Epi-ADM are administered in combination by IC50 ratio (1:2), 60 μM of wogonin pharmaceutical composition and Epi-ADM+
Drug combination has synergistic effect in 0.8 μM to 1.69 μM+0.02 μM of end level of dosage range.Wogonin pharmaceutical composition
Be administered in combination with taxol by IC50 ratio (1:4), wogonin pharmaceutical composition and taxol 9.53 μM of+3.91nM extremely
Drug combination has synergistic effect in the dosage range of 4.95 μM of+2.03nM of end level.
2.2 wogonin pharmaceutical compositions (1 group of the embodiment of the present invention) and chemotherapy drugs in combination are applied to vitro culture of human
The growth inhibition effect of liver cancer SMMC-7721 tumour cell
Wogonin pharmaceutical composition and chemotherapeutic drug Paclitaxel (PTX), Epi-ADM (EPB) use in conjunction is to external training
Support the growth inhibition effect of human liver cancer SMMC-7721, the results showed that, wogonin pharmaceutical composition and taxol press IC50 ratio
(1:3) is administered in combination, in wogonin pharmaceutical composition and taxol at 6.74 μM+0.34 μM to 50 μM+2.5 μM of end level
Dosage range in drug combination have synergistic effect.Wogonin pharmaceutical composition and Epi-ADM are joined by IC50 ratio (1:4)
Administration is closed, in wogonin pharmaceutical composition and taxol at 42.0 μM+1.02 μM to 13.74 μM+0.33 μM of end level
Drug combination has synergistic effect in dosage range.
2.3 wogonin pharmaceutical compositions (1 group of the embodiment of the present invention) are to human gastric cancer BGC-823 nude mouse xenograft tumor
Inhibiting effect
Injection wogonin pharmaceutical composition has significant growth inhibition to make human gastric cancer BGC-823 Nude Mice
With.Injection wogonin pharmaceutical composition is administered one with 60mg/kg, 30mg/kg, 15mg/kg intravenous injection administration every other day
Secondary, after administration 21 days, the T/C (%) to human gastric cancer BGC-823 Nude Mice is respectively 37.22%, 51.53%,
88.26% (first result) and 34.22%, 47.90%, 78.07% (second batch result), inhibitory rate 62.84%,
48.57%, 16.49% (first result) and 67.62%, 52.00%, 22.86% (second batch result).Fluorine under similarity condition
Uracil with when 20mg/kg tail vein injection to the T/C (%) of human gastric cancer BGC-823 Nude Mice be 31.43% (first
Batch result) and 27.92% (second batch result), tumour inhibiting rate be 70.38% and 72.10% (second batch result).Experiment knot twice
Fruit is close.The result shows that wogonin pharmaceutical composition is compared with positive drug 5-FU, tumor killing effect is close, but to experimental animal
Weight influence it is smaller.
2.4 wogonin pharmaceutical compositions (1 group of the embodiment of the present invention) are to human liver cancer SMMC-7721 nude mouse xenograft tumor
Inhibiting effect
Injection wogonin pharmaceutical composition has significant growth inhibition to human liver cancer SMMC-7721 Nude Mice
Effect.Injection wogonin pharmaceutical composition is administered one with 60mg/kg, 30mg/kg, 15mg/kg intravenous injection administration every other day
Secondary, after administration 21 days, the T/C (%) to human liver cancer SMMC-7721 Nude Mice is respectively 34.91%, 55.70%,
85.77% (first result) and 34.75%, 49.12%, 79.12% (second batch result), inhibitory rate 68.50%,
49.29%, 23.88% (first result) and 63.48%, 45.93%, 22.61% (second batch result).Fluorine under similarity condition
Uracil is to be 28.46% (the to the T/C (%) of human liver cancer SMMC-7721 Nude Mice when 20mg/kg tail vein injection
A collection of result) and 24.31% (second batch result), tumour inhibiting rate is 75.41% (first result) and 71.30% (second batch knot
Fruit).Experimental result is close twice.The result shows that wogonin pharmaceutical composition is compared with positive drug 5-FU, tumor killing effect phase
Closely, but on the weight of experimental animal it influences smaller.
Suppression of the 2.5 wogonin pharmaceutical compositions (1 group of the embodiment of the present invention) to human lung cancer H460 nude mouse xenograft tumor
Production is used
This experiment establishes Non-small cell lung carcinoma H460 nude mouse xenograft tumor model, is infused using the model evaluation
Penetrate the anti-tumor activity with wogonin pharmaceutical composition.The experimental results showed that injection wogonin pharmaceutical composition is to people
Non-small cell lung cancer H460 Nude Mice has significant growth inhibition effect.Injection wogonin pharmaceutical composition with
60mg/kg, 30mg/kg, 15mg/kg intravenous injection administration, are administered once every other day, after administration 21 days, to Non-small cell lung carcinoma
The T/C (%) of H460 Nude Mice is respectively 34.78%, 48.33%, 76.53% (first result) and 35.09%,
54.71%, the Chinese
3 toxicologic studies
3.1, influence and Chinese radix scutellariae of the wogonin pharmaceutical composition to the hemopoietic system of normal mouse and tumor-bearing mice
Influence of the plain pharmaceutical composition to the immune system of normal mouse
The results show that hemopoietic system of the wogonin pharmaceutical composition intravenous injection (60,30mg/kg) to normal mouse,
Including leucocyte in peripheral blood, bonemarrow nucleated cells number is all had no significant effect, while also not finding the siberian crabapple to normal mouse
System includes that thymus index and index and spleen index have a significant effect.Wogonin pharmaceutical composition is injected intravenously (60,30mg/kg) to lotus
Total white blood cells also have no significant effect in tumor mouse peripheral blood.
3.2, the acute toxicity test of injection wogonin pharmaceutical composition is studied
Healthy cleaning grade SD rat continuous 3 intravenous injections (iv) in 8 hours give injection wogonin medicine group
Object 150mg/ (kg. time) dosage is closed, 450mg/kg dosage is added up, except occurring being short of breath in 60 minutes, close after being administered every time
Eye lies prone and crouches outside the toxicities such as motionless, diuresis, activity reduction, does not cause animal dead.Healthy cleaning grade ICR mouse single is quiet
Injection wogonin pharmaceutical composition 250mg/kg dosage is given in arteries and veins injection (iv), occurs exhaling in 3 hours upon administration
Suction is rapid, closes one's eyes, the toxicities such as motionless, movable reduction that crouch of lying prone, and symptom is similar to rat, does not also cause animal dead.At this
Under the conditions of test dose, rat intravenous injection (iv) is administered in injection wogonin pharmaceutical composition maximum tolerated dose
Are as follows: 450mg/kg (divides 3 times and gives, be every time 150mg/kg)；To the maximum tolerated dose of mouse mainline (iv) administration are as follows:
250mg/kg。
3.3 injection wogonin pharmaceutical composition three months toxicity tests of rat
SD rat continuous three months, tail vein injection gave injection wogonin pharmaceutical composition 120,60 and respectively
30mg/ (kg.d) dosage, has apparent influence to the respiratory system of rat and motor function during administration.It is mainly shown as every
Reduced at once to appearance activity in 30 minutes after secondary administration, lie prone crouch motionless, weakness of limbs (collapse from physical exhaustion shape), be short of breath, close one's eyes, more
Urine reaction, and cause 27% animal dead, there is dosage for high, middle dosage in side reaction, and dead dosage appears in high dose, and
Low dosage and control group do not occur any abnormal response during entire test.
Three dosage groups of pharmaceutical composition of the present invention increase SD rat body weight, ingest, drinking water all has no significant effect, administration
One and a half months (administration mid-term), three months (being discontinued next day) and inspection in one month (convalescence terminates) of being discontinued, to hematology, internal organs
Weight coefficient and internal organs pathology are substantially seen and are also had no significant effect, substantially identical as control group.To the individual blood of three dosage groups
Biochemical index (CK) has raising to influence.Each phase biopsy and the discovery of midway rat cadavers histopathologic examination, to high dose group
Rat heart has certain detrimental effect, is mainly shown as myofibrosis cordis, Stromal fibroblasts, histocyte and a small amount of
The inflammatory cell infiltration fibrosis different with degree simultaneously, there are still but degree is mitigated month lesion that is discontinued.In and,
Similar pathological change is not found in the dissect of low dosage in each stage, three dosage groups are to other organs tissue without obvious pathology
Change, it is similar to control group.Under the conditions of this test dose, the injection Chinese that the continuous three months tail vein injections of SD rat are given
Baicalein pharmaceutical composition, main toxicity be activity reduce, lie prone crouch motionless, weakness of limbs, be short of breath, close one's eyes, is more
The raising of urine reaction and cretinephosphokinase (CK).Poisoning target organ is heart, and there are still but degree for month lesion that is discontinued
Mitigated.Dosage of being poisoned to death is 120mg/ (kg.d), and it is 60mg/ (kg.d), safety (nontoxic) that dosage, which occurs, in toxic reaction
Dosage is 30mg/ (kg.d).
In conclusion the wogonin pharmaceutical composition that the present invention develops is to human gastric cancer MGC-803 tumour cell, human lung cancer
H460 nude mouse xenograft tumor, liver cancer SMMC-7721 tumour cell, human gastric cancer BGC-823 nude mouse xenograft tumor, human liver cancer
SMMC-7721 nude mouse xenograft tumor etc. has apparent growth inhibition effect, this also indicates that the tool of pharmaceutical composition of the present invention
There is significant curative effect.Its toxicological experiment shows the application wogonin pharmaceutical composition to white thin in tumor-bearing mice peripheral blood
Born of the same parents' sum also has no significant effect, and the application pharmaceutical composition does not occur any abnormal response to mouse.This also illustrates, the application medicine
Compositions have the advantages that toxic side effect is small.
Claims (9)
1. a kind of wogonin pharmaceutical composition for treating neoplastic disease, which is characterized in that described pharmaceutical composition includes following weight
Measure the component of number proportion: 10~40 parts of the wogonin, 20~200 parts of cosolvent, 20~60 parts of framework material, injection
2000~3000 parts of water.
2. pharmaceutical composition as described in claim 1, which is characterized in that described pharmaceutical composition includes that following parts by weight are matched
The component of ratio: 20~30 parts of the wogonin, 80~120 parts of cosolvent, 30~50 parts of framework material, water for injection 2200~
2800 parts.
3. pharmaceutical composition as claimed in claim 1 or 2, which is characterized in that cosolvent is carbonic acid in described pharmaceutical composition
One of hydrogen sodium, glycerol, sodium phosphate, L-arginine, citric acid are a variety of.
4. pharmaceutical composition as claimed in claim 1 or 2, which is characterized in that framework material described in described pharmaceutical composition
For one of lactose, glucose, sorbierite, mannitol or a variety of.
5. pharmaceutical composition as claimed in claim 1 or 2, which is characterized in that described pharmaceutical composition includes following parts by weight
The component of number proportion is made: 25 parts of the wogonin, 100 parts of L-arginine, 40 parts of mannitol, 2500 parts of water for injection.
6. pharmaceutical composition as claimed in claim 1 or 2, which is characterized in that the dosage form of described pharmaceutical composition is freeze-dried powder
Or injection.
7. the preparation method of pharmaceutical composition as claimed in claim 1 or 2, which is characterized in that the preparation method include with
Lower processing step:
(1) after, mixing wogonin and cosolvent, add a small amount of water for injection wet, water for injection stirring, which is added, keeps drug whole
Dissolution, obtains mixed solution；
(2), take the mixed solution of step (1) to add framework material to be stirred to dissolve, active carbon is added and stirs depyrogenation, take off charcoal and take out
Filter adds injection appropriate amount of water constant volume to configure；Again through membrane filtration degerming, filtering liquid medicine is obtained, it is freeze-dried at freeze-dried powder, i.e.,
It can.
8. the preparation method of pharmaceutical composition as claimed in claim 7, which is characterized in that the preparation method processing step is (2)
In freeze drying process condition are as follows: in cryogenic temperature be -60~-40 DEG C of 3~6h of cooling time；And maintain cryogenic temperature -30
~-0 DEG C, 10~20pa of vacuum degree, cooling time is 15~30h；Continue to keep 10~20pa of vacuum degree, cryogenic temperature 0~20
DEG C, cooling time is 4~8h；Then 25~35 DEG C, 5~8h of drying time are gradually heated to, vacuum tamponade can must be lyophilized
Powder.
9. the preparation method of pharmaceutical composition as claimed in claim 8, which is characterized in that the preparation method technique is in nitrogen
It is carried out under environment.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1556101A (en) * | 2003-12-31 | 2004-12-22 | 中国药科大学 | Extraction technology of Hanbaicalein, medicinal composition and preparation technology of medicine |
| CN1562147A (en) * | 2004-03-24 | 2005-01-12 | 杭州华东医药集团生物工程研究所有限公司 | Radix scutellariae extract freeze-dried powder injection and its preparing method |
| TW201311263A (en) * | 2011-09-05 | 2013-03-16 | Univ Nat Taiwan Normal | Wogonin-containing pharmaceutical composition for inhibiting cancer stem cells growth and application thereof |
| AU2015255189A1 (en) * | 2010-03-12 | 2015-11-26 | Allergan Industrie Sas | A fluid composition comprising a hyaluronan polymer and mannitol for improving skin condition |
-
2017
- 2017-06-07 CN CN201710421158.XA patent/CN108992672B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1556101A (en) * | 2003-12-31 | 2004-12-22 | 中国药科大学 | Extraction technology of Hanbaicalein, medicinal composition and preparation technology of medicine |
| CN1562147A (en) * | 2004-03-24 | 2005-01-12 | 杭州华东医药集团生物工程研究所有限公司 | Radix scutellariae extract freeze-dried powder injection and its preparing method |
| AU2015255189A1 (en) * | 2010-03-12 | 2015-11-26 | Allergan Industrie Sas | A fluid composition comprising a hyaluronan polymer and mannitol for improving skin condition |
| TW201311263A (en) * | 2011-09-05 | 2013-03-16 | Univ Nat Taiwan Normal | Wogonin-containing pharmaceutical composition for inhibiting cancer stem cells growth and application thereof |
Non-Patent Citations (1)
| Title |
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| 王婷婷 等: ""汉黄芩素单药及联合用药对人胃癌的生长抑制作用研究"", 《第二届中国女医师大会论文集 肿瘤》 * |
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