CN109731031A - Application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics - Google Patents
Application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics Download PDFInfo
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- CN109731031A CN109731031A CN201910083972.4A CN201910083972A CN109731031A CN 109731031 A CN109731031 A CN 109731031A CN 201910083972 A CN201910083972 A CN 201910083972A CN 109731031 A CN109731031 A CN 109731031A
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- radix astragali
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Abstract
The present invention relates to the new application of Radix Astragali flavone extract, in particular to application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics.The chemotherapeutics is made of the ethanol extract of Radix Astragali with cis-platinum, and the mass ratio of the ethanol extract of Radix Astragali and cis-platinum is not less than 6:1.Laryngocarcinoma is to be only second to the high-incidence tumour of the second largest respiratory tract of lung cancer; it is high-incidence in Northern Part of China; it is cis-platinum that laryngocarcinoma chemotherapy I class, which recommends drug; however the Nephrotoxicity of cisplatin dose dependence strongly limits its clinical application; and not only tumor suppression ability enhances the present invention, while Radix Astragali flavone has good protective effect to renal damage caused by cis-platinum.
Description
Technical field
The present invention relates to the new applications of Radix Astragali flavone extract, in particular to Radix Astragali flavone extract is in preparation laryngocarcinoma chemotherapy
Application in drug.
Background technique
Laryngocarcinoma is to be only second to the high-incidence tumour of the second largest respiratory tract of lung cancer, laryngocarcinoma chemotherapy I class high-incidence in Northern Part of China
Recommendation drug is cis-platinum, however the Nephrotoxicity of cisplatin dose dependence strongly limits its clinical application.Cis-platinum mainly leads to
Peroxidating stress, injury of mitochondria and the effects of inflammatory reaction damage proximal renal tubular epithelial cells and cause acute or chronic kidney
Functional failure.
Radix Astragali is one of parts of generic medicinal plants, first recorded in Shennong's Herbal, has invigorating qi for strengthening superficies, diuresis, pus draining and toxin expelling is held back
Sore myogenic and other effects.Document report (Song Yan etc., modern medical oncology, 2013,21 (7): 1419-1421), the controllable larynx of Radix Astragali
Cancer Cell cycle, formation G2/M phase block, and by Inhibit proliferaton and apoptosis are promoted to play its antitumaous effect.
Radix Astragali flavone constituents and its extract there is no to be used for the report of laryngocarcinoma chemotherapeutics, at present with above-mentioned effective component
Based on extract also have no the application for being used to prepare laryngocarcinoma chemotherapeutics.
Summary of the invention
In order to overcome the above the deficiencies in the prior art, the present invention provides Radix Astragali flavone extracts in preparation laryngocarcinoma chemotherapeutic
Application in object, not only tumor suppression ability enhances, while Radix Astragali flavone has good protective effect to renal damage caused by cis-platinum.
Present invention employs following technical proposals in order to achieve the above objectives:
Application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics, it is characterised in that: the chemotherapeutics is by Huang
Stilbene chromocor extract and cis-platinum form, and the mass ratio of Radix Astragali flavone extract and cis-platinum is not less than 6:1.
Further, the Radix Astragali flavone extract is the ethanol extract of Radix Astragali.
Further, the Radix Astragali flavone extract is prepared by the method for following steps:
A. the Milkvetch Root for taking 1000g-3000g, the ethyl alcohol that the volume fraction that 6-8L is added is 0-95%, which is heated to reflux, to be mentioned
It takes 1-3 times, it is 1-3 hours each, resulting extracting solution is merged, paste is concentrated under reduced pressure to obtain；
B. above-mentioned paste AB-8 large pore resin absorption column is separated, successively with water, volume fraction 10%, volume point
The ethanol elution that number is 35%, the ethanol eluate that collected volume score is 35%, it is dry to the greatest extent that eluent is concentrated under reduced pressure into solvent；
C. taking above-mentioned volume fraction is that 35% ethanol eluate concentrate is scattered in 200ml water, adds 1-3 times of volume
Ethyl acetate extracts 1-2 times, combining extraction liquid, medicinal extract is concentrated under reduced pressure to obtain, 60 DEG C of vacuum drying are at dry powder.
Further, the mass percentage content of flavones is 40-60% in the Radix Astragali flavone extract, is quality hundred
Divide ratio, contained main chemical compositions include onocerin, ononin, calycosin, calycosin -7-O- β-D-
Glucopyranose, 7,2 '-dihydroxy -3 ', 4 '-dimethoxy isoflavans, 2 '-hydroxyls -3 ', 4 '-dimethoxy isoflavan -7-O-
β-D- glucopyranose, 3- hydroxyl -9,10- dimethoxy red sandalwood alkane, 9,10- dimethoxy red sandalwood alkane -3-O- β-D- pyrans Portugal
Grape sugar.
Using the subcutaneous tumor-bearing mice of laryngocarcinoma as pharmacodynamics model, the chromocor extract and cis-platinum of various dose are given respectively.Tool
Body dosage are as follows:
(1) low dose group that Radix Astragali flavone extract and cis-platinum mass ratio are 6:1: 6mg/kg chromocor extract, 1 time/d；
1mg/kg cis-platinum, 1 time/d, successive administration 14d；
(2) high dose group that Radix Astragali flavone extract and cis-platinum mass ratio are 12:1: 12mg/kg chromocor extract, 1 time/
d；1mg/kg cis-platinum, 1 time/d, successive administration 14d；
Specific administration mode is intraperitoneal injection.
Experimental study proves the chemotherapeutics of Radix Astragali flavone extract and cis-platinum composition, the enhancing of tumor suppression ability in animal body；
It finds that Radix Astragali flavone extract can increase effective weight of cisplatin chemotherapy tumor-bearing mice simultaneously, improves the existence of tumor-bearing mice
State；Can injury of kidney caused by antagonism cisplatin chemotherapy, have attenuation to cisplatin chemotherapy.
Detailed description of the invention
Fig. 1 is each group tumour growth volume change after being administered in embodiment 4；
Fig. 2 is each administration group tumour figure in embodiment 4；
Fig. 3 is each group changes of weight after being administered in embodiment 4；
Fig. 4 is each group kidney organ index in embodiment 4；
Fig. 5 is each group Pathological figure in embodiment 4.
Specific embodiment
It is further described with reference to the accompanying drawings and examples:
Embodiment 1
Application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics, the chemotherapeutics is by Radix Astragali flavone extract
It is formed with cis-platinum, and the mass ratio of Radix Astragali flavone extract and cis-platinum is not less than 6:1.The Radix Astragali flavone extract is Radix Astragali
Ethanol extract.The mass percentage content of flavones is 40% in the Radix Astragali flavone extract, is mass percent, contained
Some main chemical compositions include onocerin, ononin, calycosin, calycosin -7-O- β-D- glucopyra
Sugar, 7,2 '-dihydroxy -3 ', 4 '-dimethoxy isoflavans, 2 '-hydroxyls -3 ', 4 '-dimethoxy isoflavan -7-O- β-D- pyrans
Glucose, 3- hydroxyl -9,10- dimethoxy red sandalwood alkane, 9,10- dimethoxy red sandalwood alkane -3-O- β-D- glucopyranose.It is described
Radix Astragali flavone extract be prepared by the method for following steps:
A. 3000g Milkvetch Root is taken, is added 8L water heating and refluxing extraction 3 times, 1 hour every time, resulting extracting solution is closed
And paste is concentrated under reduced pressure to obtain；
B. above-mentioned paste AB-8 large pore resin absorption column is separated, successively with water, volume fraction 10%, volume point
The ethanol elution that number is 35%, the ethanol eluate that collected volume score is 35%, it is dry to the greatest extent that eluent is concentrated under reduced pressure into solvent；
C. taking above-mentioned volume fraction is that 35% ethanol eluate concentrate is scattered in 200ml water, adds the second of 1 times of volume
Acetoacetic ester extracts 2 times, combining extraction liquid, medicinal extract is concentrated under reduced pressure to obtain, 60 DEG C of vacuum drying are at dry powder.
Embodiment 2
Application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics, the chemotherapeutics is by Radix Astragali flavone extract
It is formed with cis-platinum, and the mass ratio of Radix Astragali flavone extract and cis-platinum is not less than 6:1.The Radix Astragali flavone extract is Radix Astragali
Ethanol extract.The mass percentage content of flavones is 50% in the Radix Astragali flavone extract, is mass percent, contained
Some main chemical compositions include onocerin, ononin, calycosin, calycosin -7-O- β-D- glucopyra
Sugar, 7,2 '-dihydroxy -3 ', 4 '-dimethoxy isoflavans, 2 '-hydroxyls -3 ', 4 '-dimethoxy isoflavan -7-O- β-D- pyrans
Glucose, 3- hydroxyl -9,10- dimethoxy red sandalwood alkane, 9,10- dimethoxy red sandalwood alkane -3-O- β-D- glucopyranose.It is described
Radix Astragali flavone extract be prepared by the method for following steps:
A. 1000g Milkvetch Root is taken, ethyl alcohol heating and refluxing extraction 2 times that 8L volume fraction is 20% are added, 2 is small every time
When, resulting extracting solution is merged, paste is concentrated under reduced pressure to obtain；
B. above-mentioned paste AB-8 large pore resin absorption column is separated, successively with water, volume fraction 10%, volume point
The ethanol elution that number is 35%, the ethanol eluate that collected volume score is 35%, it is dry to the greatest extent that eluent is concentrated under reduced pressure into solvent；
C. taking above-mentioned volume fraction is that 35% ethanol eluate concentrate is scattered in 200ml water, adds the second of 3 times of volumes
Acetoacetic ester extracts 1 time, combining extraction liquid, medicinal extract is concentrated under reduced pressure to obtain, 60 DEG C of vacuum drying are at dry powder.
Embodiment 3
Application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics, the chemotherapeutics is by Radix Astragali flavone extract
It is formed with cis-platinum, and the mass ratio of Radix Astragali flavone extract and cis-platinum is not less than 6:1.The Radix Astragali flavone extract is Radix Astragali
Ethanol extract.The mass percentage content of flavones is 60% in the Radix Astragali flavone extract, is mass percent, contained
Some main chemical compositions include onocerin, ononin, calycosin, calycosin -7-O- β-D- glucopyra
Sugar, 7,2 '-dihydroxy -3 ', 4 '-dimethoxy isoflavans, 2 '-hydroxyls -3 ', 4 '-dimethoxy isoflavan -7-O- β-D- pyrans
Glucose, 3- hydroxyl -9,10- dimethoxy red sandalwood alkane, 9,10- dimethoxy red sandalwood alkane -3-O- β-D- glucopyranose.It is described
Radix Astragali flavone extract be prepared by the method for following steps:
A. the Milkvetch Root for taking 1000g, the volume fraction that 8L is added is 95% ethyl alcohol heating and refluxing extraction for 3 times, every time 1
Hour, resulting extracting solution is merged, paste is concentrated under reduced pressure to obtain；
B. above-mentioned paste AB-8 large pore resin absorption column is separated, successively with water, volume fraction 10%, volume point
The ethanol elution that number is 35%, the ethanol eluate that collected volume score is 35%, it is dry to the greatest extent that eluent is concentrated under reduced pressure into solvent；
C. taking above-mentioned volume fraction is that 35% ethanol eluate concentrate is scattered in 200ml water, adds the second of 2 times of volumes
Acetoacetic ester extracts 2 times, combining extraction liquid, medicinal extract is concentrated under reduced pressure to obtain, 60 DEG C of vacuum drying are at dry powder.
Embodiment 4
The subcutaneous tumor-bearing mice pharmacodynamic study of laryngocarcinoma
1. laryngocarcinoma nude mice by subcutaneous xenograft tumor models construct:
(1) cell origin: Hep2 cell line.
(2) animal origin: BALB/c nude mouse (SPF grades), 4 week old, female.Purchased from Beijing biological section, dimension tonneau China
Skill limited liability company.Raising and SPF environment, environment temperature and humidity are suitable for.
(3) modeling method: taking logarithmic phase Hep2 cell, and after PBS washing, pancreatin digestion is added, and 1000rpm is centrifuged 5min,
Supernatant is abandoned, serum free medium is added, cell is resuspended, cell counter counts, required cell suspending liquid concentration about 2 × 107
A/mL.100ul cell suspension is drawn, mouse oxter, every mouse inoculation about 2 × 10 are inoculated in6A cell, inoculation volume need
Unanimously.Knurl naked eyes can be seen below, and carry out cubing (V=1/2ab to it using digital display calliper2), record knurl major diameter and short
Diameter.
(4) experimental group:
It is divided into 5 groups.Every group 6.
Blank group: not modeling is not administered；
Model group: modeling is not administered；
Cis-platinum is administered alone group: cis-platinum is successfully given in modeling, 1mg/kg, and 1 time/d；
The low dose group that Radix Astragali flavone extract and cis-platinum mass ratio are 6:1: 6mg/kg astragalus flavonid extract, 1 time/
d；1mg/kg, cis-platinum, 1 time/d；
The high dose group that Radix Astragali flavone extract and cis-platinum mass ratio are 12:1: 12mg/kg astragalus flavonid extract, 1
Secondary/d；1mg/kg, cis-platinum, 1 time/d；
Successive administration 14d, administration mode are intraperitoneal injection.
Be discontinued next day execution animal, first weighs, completely takes subcutaneous tumor mass afterwards, weighs, each group nephridial tissue is taken to make paraffin
Slice carries out light microscopic pathological study.
(5) statistical method: all data indicate that comparison among groups are examined using t with Mean ± SD, and p value < 0.05 is poor
It is different significant, it is statistically significant.
2. result
(1) each group tumour growth volume change after being administered
During groups of animals administration, every other day tumour is measured, records knurl major diameter and minor axis.Major diameter is denoted as
A, minor axis are denoted as b, utilize gross tumor volume calculation formula V=1/2ab2Each group tumor volume change after being administered is calculated, as a result as schemed
1.Compared with cis-platinum is administered alone group, growth of the chemotherapeutics of Radix Astragali flavone extract and cis-platinum composition to nude mice by subcutaneous tumour
There is apparent inhibiting effect, wherein Radix Astragali flavone extract and the high dose group effect that cis-platinum mass ratio is 12:1 are best.
(2) the standard of curative effect evaluation
The inhibiting rate of tumour=(1-T/C) × 100%
T: administration group average knurl weight；C: control group average knurl weight
Intraperitoneal administration totally 14 days, there is more apparent antitumor activity to human laryngeal cancer transplantable tumor, see Fig. 2, be to administration group in table 1
Tumor control rate variation.Wherein, independent cisplatin administration group tumour inhibiting rate is 42.15%, with the poor different significant (p of model control group
< 0.05)；The high dose combination with cisplatin group tumour inhibiting rate that Radix Astragali flavone extract and cis-platinum mass ratio are 12:1 is 62.74%, with
Model control group is poor different significant (p < 0.01)；The low dose group tumor suppression that Radix Astragali flavone extract and cis-platinum mass ratio are 6:1
Rate is 57.82%, poor different significant (p < 0.05) with model control group；
1 each group tumor control rate table (n=6, Mean ± SD) of table
(3) changes of weight
During groups of animals administration, every other day the weight of animal is monitored, records weight data.Draw weight
Variation diagram (Fig. 3).Independent cisplatin administration group mouse weight significantly reduces, compared with independent cisplatin administration group, use Radix Astragali flavone
The chemotherapeutics processing nude mice weight that extract is formed with cis-platinum increases.Illustrate that Radix Astragali flavone extract can increase cisplatin chemotherapy
Effective weight of tumor-bearing mice；Improve the survival condition of tumor-bearing mice.
(4) to the protective effect of injury of kidney
Be discontinued next day execution animal, and dissection takes each group nephridial tissue, makees paraffin section after weighing and carry out light microscopic histopathology
Observation.It draws each group kidney organ index column diagram (Fig. 4), finds compared with model group, the kidney that cis-platinum is administered alone group is dirty
Device index significantly increases (p < 0.05), and when using the chemotherapeutics of Radix Astragali flavone extract and cis-platinum composition, organ index is returned
It adjusts, and the high dose group that Radix Astragali flavone extract and cis-platinum mass ratio are 12:1 adjusts back degree significantly (p < 0.05).
And each group renal pathological findings are shown: cis-platinum is administered alone group and compared with blank control group, and renal tubular epithelial is thin
To severe hydropic degeneration in born of the same parents, tubule basophilla becomes and protein cast is common, increases in fibroblast to severe fibroblast
Life, interstitial severe cell infiltration, part Tubular epithelial cell atrophy, lumen become larger, and nephropathy degree is obvious, and Radix Astragali flavone mentions
The chemotherapeutics for taking object and cis-platinum to form can improve the damage of nephropathy caused by cis-platinum is administered alone and doses rely on.
Claims (4)
1. application of the Radix Astragali flavone extract in preparation laryngocarcinoma chemotherapeutics, it is characterised in that: the chemotherapeutics is by Radix Astragali
Chromocor extract and cis-platinum form, and the mass ratio of Radix Astragali flavone extract and cis-platinum is not less than 6:1.
2. according to right want 1 described in Radix Astragali flavone extract preparation laryngocarcinoma chemotherapeutics in application, it is characterised in that: institute
State the ethanol extract that Radix Astragali flavone extract is Radix Astragali.
3. according to right want 2 described in Radix Astragali flavone extract preparation laryngocarcinoma chemotherapeutics in application, it is characterised in that: institute
The Radix Astragali flavone extract stated is prepared by the method for following steps:
A. the Milkvetch Root for taking 1000g-3000g, the volume fraction that 6-8L is added is the ethyl alcohol heating and refluxing extraction 1-3 of 0-95%
It is secondary, it is 1-3 hours each, resulting extracting solution is merged, paste is concentrated under reduced pressure to obtain；
B. above-mentioned paste AB-8 large pore resin absorption column is separated, is successively with water, volume fraction 10%, volume fraction
35% ethanol elution, the ethanol eluate that collected volume score is 35%, it is dry to the greatest extent that eluent is concentrated under reduced pressure into solvent；
C. taking above-mentioned volume fraction is that 35% ethanol eluate concentrate is scattered in 200ml water, adds the acetic acid of 1-3 times of volume
Ethyl ester extracts 1-2 times, combining extraction liquid, medicinal extract is concentrated under reduced pressure to obtain, 60 DEG C of vacuum drying are at dry powder.
4. wanting application of the 3 Radix Astragali flavone extracts in preparation laryngocarcinoma chemotherapeutics according to right, it is characterised in that: described
Radix Astragali flavone extract in flavones mass percentage content be 40-60%, be mass percent, contained primary chemical
Ingredient includes onocerin, ononin, calycosin, calycosin -7-O- β-D- glucopyranose, 7,2 '-dihydroxies
Base -3 ', 4 '-dimethoxy isoflavans, 2 '-hydroxyls -3 ', 4 '-dimethoxy isoflavan -7-O- β-D- glucopyranoses, 3- hydroxyl
Base -9,10- dimethoxy red sandalwood alkane, 9,10- dimethoxy red sandalwood alkane -3-O- β-D- glucopyranose.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108042603A (en) * | 2018-01-16 | 2018-05-18 | 山西大学 | Radix Astragali flavone extract is preparing the application in treating nephrotic syndrome drug |
| CN108159109A (en) * | 2018-01-16 | 2018-06-15 | 山西大学 | Radix Astragali flavone extract is preparing the application in treating Neuroleptic Leukocytopenia disease drug |
-
2019
- 2019-01-29 CN CN201910083972.4A patent/CN109731031A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108042603A (en) * | 2018-01-16 | 2018-05-18 | 山西大学 | Radix Astragali flavone extract is preparing the application in treating nephrotic syndrome drug |
| CN108159109A (en) * | 2018-01-16 | 2018-06-15 | 山西大学 | Radix Astragali flavone extract is preparing the application in treating Neuroleptic Leukocytopenia disease drug |
Non-Patent Citations (2)
| Title |
|---|
| 宋岩，等: "黄芪对喉癌细胞增殖和凋亡的影响", 《实用药物与临床》 * |
| 宋岩，等: "黄芪联合顺铂对喉癌Hep-2细胞的体外抑制作用", 《中国医师杂志》 * |
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Application publication date: 20190510 |