CN102499918B - Anti-radiation medicine and manufacturing method and application - Google Patents
Anti-radiation medicine and manufacturing method and application Download PDFInfo
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Abstract
The invention belongs to the medicine technical field, and in particular relates to an anti-radiation medicine in protection of human lung embryo fibroblast and radiation injury in experimental mice. The anti-radiation medicine has a structure of 4-methoxy benzylidene-2-substituted-5(4H)-oxazolone and derivatives thereof and can be expressed in structural formula (A). The invention has the advantages that compound (A) has a significant protection effect for DNA and genome injury to human lung embryo fibroblast (MRC-5) and experimental mice caused by ionizing radiation, such as X-rays, carbon ions, etc., and non-ionizing radiation, such as ultraviolet rays and microwaves; relative survival percentage of cells irradiated by said rays can be increased significantly; DNA injury can be reduced; instability of genome is reduced; MDA (malondialdehyde) yield of mice viscera can be reduced; and SOD (Superoxide Dismutase) activity of mice viscera can be enhanced. The invention has low toxicity, wide application range and good application prospect.
Description
Technical field
The invention belongs to medical technical field, be specifically related to a kind of for the protective effect of radiation-resistant medicine to people's lung embryo fibroblast, experiment mice radiation damage.
Background technology
The occurring in nature temperature is at absolute temperature all objects above zero, and all with form of electromagnetic wave outside transfer of heat ceaselessly, this energy-delivering mode is called radiation.Radiation is outwards diffused with the form of electromagnetic wave and particle (as alpha-particle, beta-particle etc.), be divided into ultraviolet, microwave etc. at interior Non-ionizing radiation and X ray, alpha-particle, beta-particle, heavy ion etc. in interior ionizing radiation.Radiation has at present become the fourth-largest environomental pollution source after water, air, noise.
Discover: radiation can cause radiation sickness such as brain type, visible peristalsis visible intestinal peristalsis and BM form; Radiation can cause cell transformation and dead, serious vascular reaction and hemorrhage, lethal and secondary infection in early days; Radiation causes that hematopoietic disorder, immunologic hypofunction, malignant tumor take place, form intractable cutaneous ulcer late period, and multiple organ fibrosis.
Environmental radiation is ubiquitous.In the manned space flight process, the health and lives safety of irradiation space environment serious threat spaceman; The peaceful use of nuclear energy is universal day by day, the occurrence risk surge of accidents such as nuclear leakage; The develop rapidly of nuclear medicine, the doctor contacts radiation with the patient chance increases greatly; Radiation practitioners' such as nuclear science worker, radar operator, pilot safety and health are subjected to serious threat.
Radiation damage control medicine is prevention, direct, the most effective means for the treatment of.Use before the contact radioactive substance, radioprotective is to the damage of human body in advance; Be subjected to shining the back and use, can alleviate the clinical symptoms of radiation sickness, promote to recover.
The treating radiation damage drug main will be divided into several classes such as synthetic class medicine, natural product medicine and body endogenous factors at present.
Synthetic class radioprotective compound generally has characteristics such as motivated, that effect is obvious, occupies important status in clinical and experimentation.The chemical compound of phosphorous, sulfur particularly is as cystamine, cysteamine, aminoethylisothiourea, S-2-(the amino Propylamino of 3-) ethyl phosphorothioic acid ester etc.Chemical compounds such as a series of sulfur-bearings of Fa Xianing, phosphorus can both prevent radiation damage effectively in succession, but its highly toxic drawback suppresses its clinical practice.
In addition, having heat-clearing and toxic substances removing, blood circulation promoting and blood stasis dispelling, blood yiqi, yin nourishing rises white Chinese medicine in various degree radiation resistance is all arranged, and Chinese medicine medicine source is wide, toxicity is low, make it in the research of radioprotectant, demonstrate certain potentiality, natural drug such as polysaccharide, phenols, saponins, flavonoid etc.The hemopoietic inhibition that Chinese medicine mainly produces acute radiation sickness in the process for the treatment of acute radiation sickness, immunologic function reduce and gastrointestinal dysfunction plays therapeutical effect.But the absorption of Chinese patent medicine and take effect slowly, low to the Chinese medicine degree of recognition has in the world in addition also limited its application.
Metallothionein (Metallothioneins, MT), melatonin (melatonin, MLT), mescenchymal stem cell (mesenchymal cell, MSC), the radioprotective of interleukin 11 body endogenous factors such as (IL-11) report is more, but the scope of clinical practice is little, the unstability factor is many, and the clinical practice of body endogenous factors up to now rarely has report.
The development of antiradiation drug is a global problem.Though through the effort of numerous research institutions, found some active drugs, also had many weak points as practicality.Synthetic class material is tired very high, but toxic and side effects is also more obvious; Chinese herbal medicine class natural product curative effect still mostly is compound mixture greatly certainly, and the structure of effective monomer is indeterminate, lacks more careful research.The body endogenous factors is quick on the draw, but dosage is uncertain, and long-term potential toxic and side effects is still indeterminate, and also difficult large-scale application is in clinical.Therefore, also do not find at present for the severe especially effective medicine of utmost point severe radiation injury.
In addition, the body injury that radiation causes is not single, is the combined effect of multiple radiation damage.Simple removing free radical or the raising hemopoietic function of relying on reaches the white purpose of liter, still can not solve the complicated lesion that radiation causes effectively.
So low toxicity, efficient, wide spectrum, control is had both is the research direction of following antiradiation drug.The research and development of radioprotectant and radiation therapy agent shoulder heavy responsibilities.
The antiradiation effect of chemical compound (A) is not reported so far yet.
Summary of the invention
The objective of the invention is to avoid the deficiencies in the prior art to provide a kind of for radiation-resistant medicine.Has the structure that 4-(4-methoxybenzene methylene)-2-replaces-5 (4H)-Evil (miaow) oxazolone and derivant thereof.Be used for the protective effect to people's lung embryo fibroblast, experiment mice radiation damage.
For achieving the above object, the technical scheme that the present invention takes is: a kind of for radiation-resistant medicine, it is characterized in that
General structure is:
X is O, S, R in the formula
6N;
Y is the organic group shown in the following formula in the formula:
H；C
nH
2n+1；(CH
2CH
2O)
mCH
3；
R wherein
1-R
5, R
7Relatively independent ground or be carbochain, the O (CH of H, 1～20 carbon simultaneously
2CH
2O)
nCH
3(n=0-5) carbon oxygen chain, halogen Cl, Br, F, carboxylate R
8COO, sulphonic acid ester R
9SO
2O; R
6Carbochain, O (CH for H, 1～20 carbon
2CH
2O)
nCH
3(n=0-5) carbon oxygen chain; R
8, R
9Be the carbochain of H, 1～20 carbon, single-substituted, polysubstituted benzene.
Described for radiation-resistant medicine, its main feature is that people's lung embryo fibroblast MRC-5 that X ray or carbon ion beam are caused or the radiation damage of mice have protective action.
Described for radiation-resistant medicine, its main feature is that X ray or carbon ion beam irradiation descendant lung embryo fibroblast MRC-5 free radical are had clean-up effect.
Described for radiation-resistant medicine, its main feature is that the cellular genome damage that X ray or carbon ion beam ionizing radiation are caused is had protective effect.
Described for radiation-resistant medicine, its main feature is the protective effect of people's lung embryo fibroblast MRC-5 damage that ultraviolet or microwave unionized irradiation are caused.
The invention has the beneficial effects as follows that people's lung embryo fibroblast (MRC-5) that compd A causes for Non-ionizing radiations such as ionizing radiation such as X ray, carbon ion and ultraviolet, microwaves, DNA and the genome damage of experiment mice have significant protection effect, can significantly improve cell behind the above-mentioned roentgenization relative survival rate, reduce DNA damage, reduce genomic unstability, reduce mice internal organs MDA output, strengthen mice internal organs SOD activity.Toxicity is low, and applied range has good prospects for application.
Description of drawings
Fig. 1 handles mice survival curve behind the x-ray irradiation for compound I;
Fig. 2 compound I is handled, behind the x-ray irradiation, and the content of MDA in the liver organization;
Fig. 3 compound I is handled, behind the x-ray irradiation, and total SOD activity in the liver organization;
Fig. 4 chemical compound V handles can reduce the free radical that x-ray irradiation is induced generation;
Fig. 5 chemical compound V handles the free radical that can reduce the radiation-induced generation of carbon ion beam;
Fig. 6 chemical compound V handles, mice survival curve behind the x-ray irradiation;
Fig. 7 chemical compound V handles, behind the x-ray irradiation, and the content of MDA in the liver organization;
Fig. 8 chemical compound V handles, behind the x-ray irradiation, and total SOD activity in the liver organization;
Fig. 9 compounds X-XVI handles, behind the x-ray irradiation, and cell survival rate.
The specific embodiment
Below principle of the present invention and feature are described, institute only gives an actual example and to be used for explaining the present invention, is not for restriction scope of the present invention.
Chemical compound cell experiment of the present invention is solution, and solvent is dimethyl sulfoxine (DMSO).Mouse experiment is directly to irritate stomach.X-ray irradiation device (Faxitron RX-650 type, power 100kVp), carbon ion beam radiation appliance (Lanzhou heavy ion avcceleration, 80MeV/u
12C
6+, close rate 0.5Gy/min), and the microwave exposure device (Galanz, G80Q23MSL-C2,800W).
Employed experimental technique is conventional method if no special instructions among the following embodiment.
Employed reagent all can be bought from commercial channels and obtains if no special instructions among the following embodiment.
Cell line behaviour lung embryo fibroblast MRC-5 preserves center (ATCC) available from the Unite States Standard culture; The Kunming kind white mice that this experiment is adopted, SPF secondary clean animal is provided by Lanzhou University's medical college; Dimethyl sulfoxine is available from Sigma company.
Synthesizing of embodiment 1:4-(4-methoxybenzene methylene)-2-phenyl-5 (4H)-azolactones (I)
Under nitrogen current, in abundant drying and the there-necked flask (100mL) crossed with nitrogen replacement, have Dropping funnel, condensing tube, thermometer, add hippuric acid (1.79g, 10mmol), NaHCO
3(0.40g, 4.0mmol), P-methoxybenzal-dehyde (1.36g 10mmol), slowly splashes into acetic anhydride (10mL), mixture heated to 120 ℃, holding temperature stirs 4h.After reaction finishes, place and be cooled to room temperature, reactant is poured in the water (100mL), generate yellow goal object solid, it is leached.With this crude product with hot water (90 ℃, 10mL) washing is 2 times, use then cold ethanol (0 ℃, 5mL) washing is 2 times, the vacuum drying at room temperature obtains 4-(4-methoxybenzene methylene)-2-phenyl-5 (4H)-azolactone 2.24g thus.
The structure of this chemical compound is by nuclear magnetic resonance measuring, and mass spectral analysis is determined, yield 75%～85%.
Nuclear magnetic resonance, NMR identifies that its structure is:
1H?NMR(400MHz，CDCl
3，δ，ppm)3.79(s，3H)，7.21(d，J＝8.4Hz，2H)，7.23(s，1H)，7.54(m，2H)，7.61(m，1H)，8.12(d，J＝7.2Hz，2H)，8.19(d，J＝8.4Hz，2H)
Embodiment 2: adopt the same method of embodiment 1, can obtain:
4-(benzylidene)-2-phenyl-5 (4H)-azolactones (II);
4-(4-bromobenzene methylene)-2-phenyl-5 (4H)-azolactones (III);
4-(4-acetoxyl group benzylidene)-2-phenyl-5 (4H)-azolactone (IV);
4-(4-methoxyl group-3-methoxybenzene methylene)-2-phenyl-5 (4H)-azolactones (V);
4-(4-methylbenzene methylene)-2-phenyl-5 (4H)-azolactones (VI);
4-(4-Nitrobenzol methylene)-2-phenyl-5 (4H)-azolactones (VII);
4-(3,4-dimethoxy benzylidene)-2-phenyl-5 (4H)-azolactone (VIII);
4-(4-N, N-dimethyl benzene methylene)-2-phenyl-5 (4H)-azolactones (IX)
Synthesizing of embodiment 3:4-(4-methoxybenzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactones (X)
Under nitrogen current, in abundant drying and the there-necked flask (100mL) crossed with nitrogen replacement, have Dropping funnel, condensing tube, thermometer, add (4-methoxyl group) benzoyl-glycine (2.10g, 10mmol), NaHCO
3(0.40g, 4.0mmol), P-methoxybenzal-dehyde (1.36g 10mmol), slowly splashes into acetic anhydride (10mL), mixture heated to 120 ℃, holding temperature stirred 4 hours.After reaction finishes, place and be cooled to room temperature, reactant is poured in the water (100mL), generate yellow goal object solid, it is leached.With this crude product with hot water (90 ℃, 10mL) washing is 2 times, use then cold ethanol (0 ℃, 5mL) washing is 2 times, the vacuum drying at room temperature obtains 4-(4-methoxybenzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactone thus.
The structure of this chemical compound is by nuclear magnetic resonance measuring, and mass spectral analysis determines that yield is 75%-85%.
Nuclear magnetic resonance, NMR identifies that its structure is:
1H?NMR(400MHz，CDCl
3，δ，ppm)2.35(s，3H)，3.91(s，3H)，3.97(s，3H)，7.02(d，J＝8.8Hz，2H)，7.11(s，1H)，7.13(s，1H)，7.59(dd，J1＝1.6Hz，J2＝8.4Hz，1H)，8.09(d，J＝8.8Hz，2H)，8.09(d，J＝1.6Hz，1H)
Embodiment 4: adopt the same method of embodiment 3, can obtain:
4-(4-bromobenzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactones (XI);
4-(4-acetoxyl group benzylidene)-2-(p-methoxyphenyl)-5 (4H)-azolactone (XII);
4-(4-methoxyl group-3-methoxybenzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactones (XIII);
4-(4-methylbenzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactones (XIV);
4-(4-Nitrobenzol methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactones (XV);
4-(4-N, N-dimethyl benzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactones (XVI).
Embodiment 5:4-(4-methoxybenzene methylene)-2-phenyl-5 (4H)-azolactone (I) (to call compound I in the following text) is to the radiation protection effect of people's lung embryo fibroblast (MRC-5) of X ray and microwave radiation processing damage:
1. experimental technique: the mensuration of cell relative survival rate realizes with mtt assay.Concrete grammar is as follows: 1. collect the logarithmic (log) phase cell, adjust concentration of cell suspension, be inoculated in 96 hole flat undersides according to every hole 100 μ L, it is 6000/hole (edge hole is filled with aseptic PBS) that bed board makes cell density to be measured.2.5%CO
2, 37 ℃ hatch, change the culture medium (every hole cumulative volume 200 μ L) that contains compound I next day.After 2 hours, take X ray or microwave to carry out radiation treatment.3.5%CO
2, 37 ℃ hatched 48 hours, observe under the inverted microscope.4. every hole adds 20 μ L MTT solution (5mg/mL, with aseptic PBS preparation, pH 7.4, i.e. 0.5%MTT), continues to cultivate 4h.5. stop cultivating, the careful suction removed culture fluid in the hole.6. every hole adds 150 μ L dimethyl sulfoxines, puts low-speed oscillation 10min on the shaking table, and crystal is fully dissolved.7. measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place, the record result.8. zeroing hole (culture medium, MTT, dimethyl sulfoxine), control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxine) and test hole (cell, drug solution, culture fluid, MTT, dimethyl sulfoxine) are set simultaneously.
2. experimental result
Table 1 compound I is handled the back cell survival rate
As shown in table 1, under the different compound concentrations of choosing such as 30,60,90,120,200,400,600 μ M, the relative vigor of compound I group cell is similar with the dimethyl sulfoxine group.During same concentrations, all do not have significant difference (p＞0.05) between the dimethyl sulfoxine group of compound I group and correspondence, the concentration dose point of choosing above is described, the toxicity of compound I is mainly from the solvent dimethyl sulfoxine.When being solvent with the dimethyl sulfoxine, the half lethal concentration LC50 of compound I is 300 μ M.
The survival rate of table 2 50 μ M compound I processing+x-ray irradiation cells
(annotate: this place p value is the compound I group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
As shown in table 2, the cell of matched group, dimethyl sulfoxine processed group through the 2Gy x-ray irradiation after the relative vigor of cell significantly descend (the p value is respectively 0.02 and 0.02).The dimethyl sulfoxine of this concentration (50 μ M) does not have significant protective effect (p=0.07) to radiation.But after 50 μ M compound I were handled, the relative vigor of cell was significantly higher than simple irradiation group and dimethyl sulfoxine+irradiation group (the p value is respectively 0.03,0.02), illustrates that compound I has tangible radiate protective action to cell.
The survival rate of table 3 50 μ M compound I processing+90cW microwave exposure cells
(annotate: this place p value is the compound I group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
The microwave exposure result is as shown in table 3, the cell of matched group, dimethyl sulfoxine processed group is through the relative vigor of cell behind the microwave exposure of 90cW significantly descend (the p value is respectively 0.01 and 0.03), and compound I is used microwave exposure after handling again, significantly improves cell activity (p=0.02) on the contrary.Dimethyl sulfoxine does not have significant protective effect (p=0.52) to radiation.But after 50 μ M compound I were handled, the relative vigor of cell was significantly higher than simple irradiation group and dimethyl sulfoxine+irradiation group (the p value is respectively 0.007,0.036), illustrates that compound I has significant protective effect too to microwave radiation.
Embodiment 6: the radiation protection effect that the experiment mice that compound I is handled X-radiation damages
1. experimental technique:
(1) detection of survival rate
Observe its growth and behavioral aspect, statistical analysis mice survival condition through postradiation mice.
(2) detection of organ index
The cervical vertebra dislocation method is put to death mice, dissects and wins liver, spleen and thymus, blots residual blood with filter paper, and analytical balance is weighed.The computing formula of each organ index is as follows: liver index=liver weight (g)/body weight (g); Index and spleen index=spleen weight (g)/body weight (g); Thymus index=thymic weight (g)/body weight (g).
(3) the peripheral blood cells number detects
Mouse tail is got blood 40 μ L, and the EDTA anticoagulant is checked in commune hospital clinical laboratory of Lanzhou Branch of the Chinese Academy of Sciences.
(4) detection of MDA content in the liver organization
Mice is put to death in the cervical vertebra dislocation, takes out liver organization.Make 10% tissue homogenate with normal saline, carry out each index determining.MDA assay test kit is purchased and is built up bio-engineering research institute in Nanjing.The compound method of each reagent and the assay method of every index protein content etc. all carry out according to the test kit description before the index determining.Measure the light absorption value of every index with all band spectrophotometer (Multiskan), and calculate desired data according to the formula that the test kit description provides.
Computing formula is as follows: MDA content in the liver (nmol/mg of unit)=(measure pipe OD-and measure blank pipe OD)/(the blank pipe of standard pipe OD-standard OD) * 10nmol/mL/ institute test sample product protein concentration.
(5) detection of SOD activity in the liver organization
Mice is put to death in the cervical vertebra dislocation, takes out liver organization.Make 10% tissue homogenate with normal saline, carry out each index determining.SOD determination of activity test kit is purchased and is built up bio-engineering research institute in Nanjing.The compound method of each reagent and the assay method of every index protein content etc. all carry out according to the test kit description before the index determining.With the light absorption value of every index of all band spectrophotometric determination, and the formula that provides according to the test kit description calculates desired data.
Computing formula is as follows: extension rate ÷ institute this protein content of test sample before total SOD activity (U/mg of unit) in the liver=(control tube OD-measures pipe OD)/control tube/50% * reactant liquor cumulative volume ÷ sampling amount * sample test.
2. experimental result
(1) mice survival rate
Behind the irradiation, the survival rate of mice drops to 50% time matched group and compound I group respectively at the 6th day and the 22nd day, illustrates that compound I can prolong the half death time (as Fig. 1) of irradiation mice.The mice of irritating stomach by chemical compound not irradiation group 30 days survival rate (survival rate is 100%) is compared zero difference with control group mice (survival rate is 100%), illustrates that the toxicity of medicine is very weak.In addition, behind the irradiation the 19th day, compound I group mice survival rate was respectively 60%, apparently higher than matched group (10%), illustrates that medicine can improve the mice survival rate, has certain radiate protective action to body.
(2) detection of mice organ index
Table 4: the influence that compound I changes organ index
Table 5: organ index rate of change behind the irradiation
The liver index of compound I group mice is compared with matched group and there was no significant difference, illustrates to take medicine to the not influence of liver index, and is minimum to hepatotoxicity.Behind the irradiation, control group mice liver index extremely significantly descends (p=0.0000122), compound I group mouse liver index significantly descend (p=0.0025).As shown in table 5, through irradiation, matched group liver index reduces by 32.8%, and compound I group liver index reduces by 18.1%, illustrates that medicine can effectively alleviate radiation to the damage of liver.
The index and spleen index of compound I group mice compared with the control and there was no significant difference illustrate that taking medicine does not influence index and spleen index, and is minimum to spleen toxicity.Through irradiation, each experimental mice index and spleen index (matched group: p=0.000094 that all significantly descends; Compound I group: p=0.000014).As shown in table 5, reduce by 45.3% through irradiation matched group index and spleen index, compound I group index and spleen index reduces by 50.4%, shows that medicine is little for the index and spleen index variable effect that irradiation causes.
The thymus index of medicine group mice compared with the control and there was no significant difference illustrate that taking medicine does not influence thymus index, and is minimum to thymus toxicity.Through irradiation, the control group mice thymus index significantly descends (p=0.029), and not remarkable (the compound I group: p=0.297) of compound I group mouse thymus index decreased.As shown in table 5, reduce by 28.8%, 1# medicine group through irradiation matched group thymus index and reduce by 3.97%, illustrate that two kinds of medicines all can effectively alleviate radiation to the damage of thymus.
(3) GANRA class medicine is to the influence of hemogram variation
: table 6: compound I is to the influence of mice hemogram
(p＜0.001 compared with the control for *, p＜0.05 * *; #, p＜0.05 ##, p＜0.01 is compared with the radiation contrast)
: table 7: peripheral blood cells number of variations rate behind the irradiation
Shown in table 6 and table 7, matched group is through after shining, and mice peripheral blood leucocyte, lymphocyte, neutrophilic granulocyte and number of platelets all significantly reduce.Peripheral blood leucocyte, lymphocyte, intermediate value cell (namely have a liking for acid, have a liking for alkali and mononuclear cell sum), neutrophilic granulocyte, number of platelets all are significantly higher than matched group behind the compound I irradiation; difference has statistical significance; show that compound I can weaken the blood cell count reduction that irradiation causes, the hemopoietic system damage that causes for acute exposure has the certain protection effect.
(4) compound I is to the influence of oxidation resistance in the liver organization
Compound I is to the influence of MDA content in the liver organization
As shown in Figure 2: the compound I group MDA content of irradiation significantly is not lower than matched group, shows that compound I can reduce MDA content in the liver organization.Radiant energy is induced the generation free radical, causes body generation peroxide injury, so MDA content significantly increases (matched group: p=0.0073 behind the irradiation; Compound I group: p=0.0077).
Compound I is to the influence of SOD activity in the liver organization
As shown in Figure 3: the SOD activity in the compound I group liver of irradiation does not show that a little more than matched group compound I can improve SOD activity in the liver organization.Radiant energy causes the reduction of SOD activity in the liver organization, but the SOD activity of compound I group is higher than matched group (compound I 8Gy and matched group 8Gy:p=0.27; ), illustrate that compound I can be alleviated because the active reduction of the SOD that irradiation causes is played certain radiate protective action to body.
Embodiment 7:4-(4-methoxyl group-3-methoxybenzene methylene)-2-phenyl-5 (4H)-azolactone (V) (to call chemical compound V in the following text) is to the radiation protection effect of people's lung embryo fibroblast (MRC-5) damage of X ray, carbon ion beam, ultraviolet, microwave radiation processing
1. experimental technique
The mensuration of cell relative survival rate realizes with mtt assay.Concrete grammar is as follows: 1. collect the logarithmic (log) phase cell, adjust concentration of cell suspension, be inoculated in 96 hole flat undersides according to every hole 100 μ L, it is 6000/hole (edge hole is filled with aseptic PBS) that bed board makes cell density to be measured.2.5%CO
2, 37 ℃ hatch, change the culture medium (every hole cumulative volume 200 μ L) that contains chemical compound V next day.After 2 hours, take rays such as X ray, carbon ion beam, ultraviolet, microwave to carry out radiation treatment.3.5%CO
2, 37 ℃ hatched 48 hours, observe under the inverted microscope.4. every hole adds 20 μ L MTT solution (5mg/mL, with aseptic PBS preparation, pH 7.4, i.e. 0.5%MTT), continues to cultivate 4h.5. stop cultivating, the careful suction removed culture fluid in the hole.6. every hole adds 150 μ L dimethyl sulfoxines, puts low-speed oscillation 10min on the shaking table, and crystal is fully dissolved.7. measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place, the record result.8. zeroing hole (culture medium, MTT, dimethyl sulfoxine), control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxine) and test hole (cell, drug solution, culture fluid, MTT, dimethyl sulfoxine) are set simultaneously.
2 experimental results
Table 8 chemical compound V handles the survival rate of back cell
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
As shown in table 8, under the different compound concentrations of choosing such as 25,50,75,100,200,400 μ M, the relative vigor of chemical compound V group cell is all than dimethyl sulfoxine group height.Explanation is at the above concentration dose point of choosing, and chemical compound V not only self does not almost have toxicity, and can alleviate the toxicity of solvent dimethyl sulfoxine.When chemical compound V concentration is 50 μ M, with respect to other concentration, the effect (p=0.04) of the relative vigor of obvious promotion cell is arranged.With dimethyl sulfoxine during as solvent, the half lethal concentration LC50 of chemical compound V＞400 μ M.
Table 9: the survival rate of cell behind the chemical compound V processing+x-ray irradiation
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
As shown in table 9, cellular control unit is through the relative vigor of cell behind the 2Gy x-ray irradiation significantly descend (p=0.02).There is not significant difference (p＞0.05) between variable concentrations dimethyl sulfoxine and the simple irradiation group; But; when chemical compound V concentration is respectively 25,50,75,100 μ M, with respect to the solvent dimethyl sulfoxine, the relative vigor of cell all improve (the p value is respectively 0.03,0.02,0.23,0.01); illustrate that the cell of chemical compound V has tangible radiate protective action.
The survival rate of cell behind the table 10:50 μ M chemical compound V+ carbon ion beam irradiation
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
As shown in table 10, the carbon ion beam predose added 50 μ M chemical compound V in 2 hours, and 48h measures the MTT data after the radiation of 2Gy 80keV/ μ m carbon ion beam.The result shows that chemical compound V has significant protective effect (p=0.02) for cell.And the protective effect of solvent dimethyl sulfoxine not obvious (p=0.26), chemical compound V is with respect to dimethyl sulfoxine, to the anti-radiation protection effect more obvious (p=0.0005) of cell.
: the survival rate of cell behind table 11:50 μ M chemical compound V+ ultraviolet (UVC) irradiation
(annotate: this place p value is chemical compound V with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
As shown in table 11, matched group is through 10J/m
2Ultraviolet (UVC) irradiation after the relative vigor of cell significantly descend (p=0.001); after handling, dimethyl sulfoxine processed group and chemical compound V use ultraviolet irradiation again; cell survival rate does not have significant change (the p value is respectively 0.47 and 0.36), shows that both have the certain protection effect to ultraviolet radiation.Chemical compound V and dimethyl sulfoxine do not have difference (p=0.69) under radiationless situation; But behind the ultraviolet irradiation, the cell survival rate of chemical compound V group is significantly higher than dimethyl sulfoxine group (p=0.005), illustrates that the ultraviolet radiation of chemical compound V has significant protective effect too.
The protective effect of the microwave exposure MRC-5 cell of the 90cW of table 12:50 μ M chemical compound V
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
As shown in table 12, the cell of matched group, dimethyl sulfoxine processed group is through the relative vigor of cell behind the microwave exposure of 90cW significantly descend (the p value is respectively 0.01 and 0.03), and chemical compound V uses microwave exposure, cell survival rate not to have significant change (p=0.33) after handling again.Dimethyl sulfoxine does not have significant protective effect (p=0.52) to radiation.But after 50 μ M chemical compound V handled, the relative vigor of cell was significantly higher than dimethyl sulfoxine+irradiation group (the p value is 0.045), illustrates that the microwave radiation of chemical compound V has significant protective effect too.
Embodiment 8: the clean-up effect of the X ray of chemical compound V and carbon ion beam irradiation descendant lung embryo fibroblast (MRC-5) free radical
1 experimental technique
The analysis of free radical yield adopts the DCF-DA staining to realize.Concrete operations are as follows: 1, the coverslip of cleaning is placed
In the culture dish, every ware is planted 500,000 cells.2,2h replaces former culture medium with the pastille culture medium before the radiation treatment.The X ray of 2Gy or 2Gy 80keV/ μ m
12C
6+Ion beam irradiation is handled.3, got time point 0 hour and 1 hour.Clean coverslip 2 times with serum-free medium, add 10 μ M DCF-DA dyestuffs of serum-free medium configuration afterwards.4,5%CO
2, 37 ℃ hatch 20 minutes after, clean 2 times with serum-free medium.Coverslip drips serum-free medium, tips upside down on the coverslip fluorescence microscope microscopy, 0.02s take pictures (fixedly time of exposure).
2 experimental results
As shown in Figure 4 and Figure 5, the MRC-5 cell through 2Gy X ray or 2Gy 80keV/ μ m,
12C
6+When ion beam irradiation was handled back 0h, the cell freeradical yield that matched group is handled was maximum, and fixedly time of exposure (0.02s) is taken pictures.The yield of free radical is directly proportional with fluorescence intensity.The result shows that the cell freeradical yield of matched group is maximum, and dimethyl sulfoxine takes second place, and chemical compound V is minimum.The growing amount of the overall free radical of cell that 1h cultivate to handle behind the irradiation lacks that (matched group content is maximum during obviously than 0h, the dimethyl sulfoxine group is taken second place, and chemical compound V is minimum, and the cultivation through 1h is described behind the irradiation, cell starts the free radical scavenging path, and the free radical of generation begins to be eliminated.The result shows that chemical compound V has good free radical scavenger effect, and the cell injury that produces owing to free radical is had good protective action.
Embodiment 9: the X ray of chemical compound V and carbon ion beam radiation treatment people lung embryo fibroblast (MRC-5) are induced the protection effect that produces DNA damage and genomic instability
1 experimental technique
The mensuration of radiation treatment DNA damage adopts the method for immunohistochemical staining to realize.Concrete operations are as follows: 1, sample preparation.The coverslip of cleaning is placed
In the culture dish, the cell kind under the digestion is gone into culture dish, 500,000 cells of every ware.Predose 2 hours is replaced former culture medium with containing chemical compound 5 chemical compound culture medium.0.5Gy X ray or 2Gy 80keV/ μ m,
12C
6+Ion beam irradiation is handled, and the different time sampling is fixing.2, cell is fixing.A, sucking-off culture medium are given a baby a bath on the third day after its birth time with PBS, add the paraformaldehyde that 1.5mL4% now prepares, and room temperature is placed 10min; B, sucking-off paraformaldehyde, the methanol of 1.5mL-20 ℃ of pre-cooling of adding, fixedly 20min; C, sucking-off methanol fixative add 4mL 70% ethanol ,-20 ℃ of preservations; 3, immunofluorescence dyeing A, sucking-off 70% ethanol add 1mL PBS rehydration, 10min * 3 time; 5min under the Triton X-100 room temperature of B, adding 2mL 0.5%; C, wash once with PBS, add mounting 2h under the 2mL 5% skim milk room temperature; D, sucking-off skim milk add 200 μ L γ H2AX primary antibodies, 2h under the room temperature (or 4 ℃ spend the night).E, on horizontal shaking table, wash 10min * 4 time with PBS; F, to add 200 μ L two anti-, and lucifuge, room temperature are placed 1h (or 4 ℃ spend the night).G, on horizontal shaking table, lucifuge is washed 10min * 4 time with PBS; H, DAPI dyeing: inoculation is had the one side of cell to drip to contain in right amount the DAPI De Kang temper mountant of going out, tip upside down on and clean on the microscope slide that dries; I, 4 ℃ keep in Dark Place; J, under fluorescence microscope, observe and counting the foci quantity of 100 cells of each dose point statistics.Under the immunofluorescence microscopy, each green bright spot all represents place dna double chain interruption.Bright spot should clearly become clear, be easy to differentiate.
The mensuration of radiation treatment genome stability adopts the micronucleus blocked method to realize.Concrete experimental technique is as follows: 1, the cell kind is gone in 12 orifice plates 200,000 cells in every hole.Predose 2h replaces former culture medium with the pastille culture medium.The X ray of 2Gy or 2Gy 80keV/ μ m,
12C
6+Ion beam irradiation is handled, and adds the cytochalasin B of 15 μ L, 0.25 μ g/mL after the radiation immediately.2, fixing: after continuing to cultivate 32h, outwell culture medium, wash twice with PBS, add 1mL Kano fixative, room temperature is 10min fixedly, outwells fixative, spends the night in air drying.3, dyeing: every ware adds 1mL30 μ g/mL acridine orange dyeing liquor, dyeing 10min, reclaim dyeing liquor, clean once with PBS, covered is inhaled and is abandoned unnecessary PBS, use fluorescence microscope, count the dikaryocyte number and the micronucleus sum that contain micronucleus in 800 dikaryocytes, calculate micronuclear rates, in percentage rate.Cellular morphology is intact after acridine orange dyeing, the rounded bright spot of micronucleus, and volume accounts for the 1/10-1/50 of main nuclear, sends yellow-green fluorescence, and is high-visible.
2 experimental results
By table 13 and table 14 as can be known, 1h after the 0.5Gy x-ray irradiation is handled, γ H2AX foci number is matched group the highest (9.64 ± 0.72), and the dimethyl sulfoxine group is taken second place (5.62 ± 0.52), and chemical compound V organizes minimum (2.83 ± 0.40).Compare with the 0.5Gy matched group, the p value of dimethyl sulfoxine and chemical compound V is respectively 0.063 and 0.006, and chemical compound V is 0.03 with respect to dimethyl sulfoxine group p value.Illustrate that the DNA damage of chemical compound V has significant protective effect.Behind the irradiation 1h, chemical compound V is 0.034 with respect to the p value of dimethyl sulfoxine, illustrates that chemical compound V is stronger than the protective effect of dimethyl sulfoxine.Behind the 4h, the double-strand break level of chemical compound V still is lower than matched group and dimethyl sulfoxine group (the p value is respectively 0.058 and 0.061), maintains no irradiation background level (p=0.967).
The result of carbon ion beam irradiation similarly, 1h behind the 0.5Gy irradiation, matched group γ H2AX foci number drops to 4.54 ± 0.44 after background level 3.99 ± 0.21 rises to 8.22 ± 0.36,4h.1h is with respect to matched group behind irradiation for the dimethyl sulfoxine group, and (p=0.014) obviously reduced in the double-strand break site, not variation (p=0.234) of 1h relatively behind the 4h.The protective effect of chemical compound V is very obvious, and 1h behind the irradiation, γ H2AX foci number are lower than matched group and dimethyl sulfoxine group (the p value is respectively 0.0057 and 0.041); Behind the 4h during with respect to 1h γ H2AX foci number do not have significant change (p=0.101).Radiation has more obvious protection effect to carbon ion beam for dimethyl sulfoxine for presentation of results, chemical compound V.
This shows that chemical compound V can effectively protect DNA to avoid radiation damage.
By table 15 and 16 as can be known, after cell is handled through the 2Gy x-ray irradiation, micronuclear rates (the MNF of matched group, Frequency of Micronuclei) risen to 0.491 ± 0.028 by 0.022 ± 0.031, shown that the chromosome of cell has been subjected to tangible radiation damage (the p value is respectively 0.004 and 0.006).The micronuclear rates difference not obvious (the p value is respectively 0.827 and 0.561) of matched group and dimethyl sulfoxine group and chemical compound V group when 0Gy handles.Handle through the 2Gy X-radiation, the micronuclear rates of chemical compound V is starkly lower than matched group and dimethyl sulfoxine group (the p value is respectively 0.031 and 0.007).Illustrate that chemical compound V can reduce the radiation damage of cellular genome.
After carbon ion beam spoke bundle radiation treatment, the micronuclear rates of matched group has risen to 0.248 ± 0.016 by 0.041 ± 0.004 respectively.Show that cell has been subjected to the HIB radiation and has produced tangible genome damage (the p value is respectively 0.002 and 0.002).But after 2Gy carbon ion beam radiation treatment, the micronuclear rates of chemical compound V is starkly lower than matched group and dimethyl sulfoxine group (the p value is respectively 0.018 and 0.012).Illustrate that the damage of the chemical compound V genome that radiation causes for HIB also has the certain protection effect.
: γ H2AX foci number statistical table behind the table 13:MRC-5 cell x-ray irradiation
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
: γ H2AX foci number statistical table behind the table 14:MRC-5 cell carbon ion beam irradiation
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
: table 15:X x ray irradiation x micronuclear rates statistics
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
: table 16: carbon ion beam irradiation micronuclear rates statistics
(annotate: this place p value is chemical compound V group with respect to the dimethyl sulfoxine group
*P＜0.05
*P＜0.01)
Embodiment 10: the radiation protection effect of the experiment mice damage that the X-radiation of chemical compound V is handled
1 experimental technique
(1) detection of survival rate
Observe its growth and behavioral aspect, statistical analysis mice survival condition through postradiation mice.
(2) detection of organ index
The cervical vertebra dislocation method is put to death mice, dissects and wins liver, spleen and thymus, blots residual blood with filter paper, and analytical balance is weighed.The computing formula of each organ index is as follows: liver index=liver weight (g)/body weight (g); Index and spleen index=spleen weight (g)/body weight (g); Thymus index=thymic weight (g)/body weight (g)
(3) the peripheral blood cells number detects
Mouse tail is got blood 40 μ L, and the EDTA anticoagulant is checked in commune hospital clinical laboratory of Lanzhou Branch of the Chinese Academy of Sciences.
(4) detection of MDA content in the liver organization
Mice is put to death in the cervical vertebra dislocation, takes out liver organization.Make 10% tissue homogenate with normal saline, carry out each index determining.MDA assay test kit is purchased and is built up bio-engineering research institute in Nanjing.The compound method of each reagent and the assay method of every index protein content etc. all carry out according to the test kit description before the index determining.Measure the light absorption value of every index with all band spectrophotometer (Multiskan), and calculate desired data according to the formula that the test kit description provides.
Computing formula is as follows: MDA content in the liver (nmol/mg of unit)=(measure pipe OD-and measure blank pipe OD)/(the blank pipe of standard pipe OD-standard OD) * 10nmol/mL/ institute test sample product protein concentration.
(5) detection of SOD activity in the liver organization
Mice is put to death in the cervical vertebra dislocation, takes out liver organization.Make 10% tissue homogenate with normal saline, carry out each index determining.SOD determination of activity test kit is purchased and is built up bio-engineering research institute in Nanjing.The compound method of each reagent and the assay method of every index protein content etc. all carry out according to the test kit description before the index determining.With the light absorption value of every index of all band spectrophotometric determination, and the formula that provides according to the test kit description calculates desired data.
Computing formula is as follows: extension rate ÷ institute this protein content of test sample before total SOD activity (U/mg of unit) in the liver=(control tube OD-measures pipe OD)/control tube/50% * reactant liquor cumulative volume ÷ sampling amount * sample test.
2. experimental result
(1) influence of the mice survival of chemical compound V
From shown in Figure 6 behind irradiation the survival rate of mice drop to for 50% time, matched group is the 6th day, chemical compound V group is the 26th day, illustrates that chemical compound V can prolong the half death time of irradiation mice.The mice of irritating stomach by medicine not irradiation group 30 days survival rate (chemical compound V group is 90%) is compared zero difference with control group mice (survival rate is 100%), the toxicity that medicine is described a little less than.In addition, 20 days survival rate of chemical compound V group mice is 80% behind the irradiation, apparently higher than matched group (10%), illustrates that chemical compound V can improve the mice survival rate, has certain radiate protective action to body.
(2) influence of the mice organ index of chemical compound V
Table 17: the influence that the organ index of chemical compound V changes
Organ index rate of change behind table 18 irradiation
The liver index of chemical compound V group mice is compared with matched group and there was no significant difference, illustrates to take medicine to the not influence of liver index, and is minimum to hepatotoxicity.Behind the irradiation, control group mice liver index extremely significantly descends (p=0.0000122), and chemical compound V group mouse liver index decreased is remarkable (p=0.08) not.Shown in table 17 and 18, reduce by 32.8% through irradiation matched group liver index, chemical compound V group liver index only reduces by 5.2%, illustrates that medicine can significantly effectively alleviate radiation to the damage of liver.
The index and spleen index of chemical compound V group mice compared with the control and there was no significant difference illustrate that taking medicine does not influence index and spleen index, and is minimum to spleen toxicity.Through irradiation, each experimental mice index and spleen index (matched group: p=0.000094 that all significantly descends; Chemical compound V group: p=0.00022).Shown in table 18, reduce by 45.3% through irradiation matched group index and spleen index, chemical compound V group index and spleen index reduces by 48.5%, shows that medicine is little for the index and spleen index variable effect that irradiation causes.
The thymus index of chemical compound V group mice compared with the control and there was no significant difference illustrate that taking medicine does not influence thymus index, and is minimum to thymus toxicity.Through irradiation, the control group mice thymus index significantly descends (p=0.029), and chemical compound V group mouse thymus index decreased is remarkable (p=0.058) not.Shown in table 18, reduce by 28.8% through irradiation matched group thymus index, chemical compound V group reduces by 9.35%, illustrates that two kinds of medicines all can effectively alleviate radiation to the damage of thymus.
(3) influence of the hemogram variation of chemical compound V
: table 19: the influence of the hemogram of chemical compound V
(*, p＜0.05, * *, p＜0.001 is compared with the control; #, p＜0.05, ##, p＜0.01 is compared with the radiation contrast)
Table 20: peripheral blood cells number of variations rate behind the irradiation
Shown in table 19 and table 20, matched group is through after shining, and mice peripheral blood leucocyte, lymphocyte, neutrophilic granulocyte and number of platelets all significantly reduce.Peripheral blood leucocyte, lymphocyte, intermediate value cell (namely have a liking for acid, have a liking for alkali and mononuclear cell sum), neutrophilic granulocyte, number of platelets all are significantly higher than matched group behind the chemical compound V group irradiation; difference has statistical significance; chemical compound V can weaken the blood cell count reduction that irradiation causes, the hemopoietic system damage that causes for acute exposure has the certain protection effect.
(4) influence of oxidation resistance in the liver organization of chemical compound V
A. the influence of MDA content in the liver organization of chemical compound V
As shown in Figure 7: the MDA content in the chemical compound V of the irradiation group liver organization does not show that a little less than matched group chemical compound V can reduce the MDA content of liver organization.Radiant energy induces the free radical of generation to cause body generation peroxide injury, so MDA content significantly increases (matched group: p=0.0073) behind the irradiation; And chemical compound V group MDA changes of contents little (p=0.48).MDA content increase by 80.6% in the liver organization behind the matched group irradiation; and chemical compound V group has only increased by 1.56%; the result shows; chemical compound V can significantly reduce the peroxidating product that irradiation causes---and MDA content increases; suppress because radiation-induced peroxide injury plays certain radiate protective action to body.
B. the influence of SOD activity in the liver organization of chemical compound V
As shown in Figure 8: the SOD activity in the chemical compound V group liver is significantly higher than matched group, and difference has statistical significance (p＜0.01), shows that chemical compound V can improve SOD activity in the liver organization.Radiant energy causes the reduction of SOD activity in the liver organization, but the SOD activity of chemical compound V group is higher than matched group, illustrates that chemical compound V can be alleviated because the active reduction of the SOD that irradiation causes is played certain radiate protective action to body.
More than all experiments show that compound I and V have low-down cytotoxicity.Experiments such as MTT experiment, free radical horizontal detection, micronucleus blocking experiment and the chain interruption of immunohistochemical staining detection dna double show that compound I and V have tangible protective action for ultimate death.Compound I and V have very strong radical scavenging activity; by reducing radiation-induced free radical level, weakening the radiation damage of free yl induction; effectively protection DNA avoids radiation damage, thereby avoids the generation of genomic instability, brings into play radiation-resistant effect.Mouse experiment shows that also compound I and V are for the survival of mice, and the damage of mouse thymus, liver etc. has certain radiation protection effect.In addition, compound I and V can significantly reduce the liver MDA amount, strengthen liver SOD, have good effect for mouse liver free radical clear.
Embodiment 11:4-(4-N, N-dimethyl benzene methylene)-2-(p-methoxyphenyl)-5 (4H)-azolactone (to call compounds X in the following text---XVI) to the radiation protection effect of people's lung embryo fibroblast (MRC-5) of X ray and microwave radiation processing damage
1 experimental technique
The mensuration of cell relative survival rate realizes with mtt assay.Concrete grammar is as follows: 1. collect the logarithmic (log) phase cell, adjust concentration of cell suspension, be inoculated in 96 hole flat undersides according to every hole 100 μ L, it is 6000/hole (edge hole is filled with aseptic PBS) that bed board makes cell density to be measured.2.5%CO
2, 37 ℃ hatch, change next day and contain compounds X--the culture medium (every hole cumulative volume 200 μ L) of-XVI.After 2 hours, take X ray or microwave to carry out radiation treatment.3.5%CO
2, 37 ℃ hatched 48 hours, observe under the inverted microscope.4. every hole adds 20 μ L MTT solution (5mg/mL, with aseptic PBS preparation, pH 7.4, i.e. 0.5%MTT), continues to cultivate 4h.5. stop cultivating, the careful suction removed culture fluid in the hole.6. every hole adds 150 μ L dimethyl sulfoxines, puts low-speed oscillation 10min on the shaking table, and crystal is fully dissolved.7. measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place, the record result.8. zeroing hole (culture medium, MTT, dimethyl sulfoxine), control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxine) and test hole (cell, drug solution, culture fluid, MTT, dimethyl sulfoxine) are set simultaneously.
2 experimental results
As shown in Figure 9, the cell of matched group, dimethyl sulfoxine processed group is through the relative vigor of cell behind the 2Gy x-ray irradiation significantly descend (the p value is respectively 0.002 and 0.01).The dimethyl sulfoxine of this concentration (50 μ M) does not have significant protective effect (p=0.07) to radiation.But; after 50 μ M compounds Xs are handled; the relative vigor of cell is significantly higher than simple irradiation group and dimethyl sulfoxine+irradiation group (the p value is respectively 0.03,0.02); the survival rate P value that cell survival rate was crossed cell with respect to dmso treatment after compounds X I, XII, XIII, XIV, XV, XVI handled is respectively 0.05,0.02,0.03,0.01,0.46,0.09; proof compounds X, XI, XII, XIII, XIV species chemical compound can obviously improve the survival rate of cell behind the irradiation, illustrate that this compounds has tangible radiate protective action to cell.
Infer thus the compd A class totally 16 kinds of chemical compounds also should have similar radiation damage protection effect.
The above only is preferred embodiment of the present invention, not in order to limiting the present invention, all any modifications of doing within the spirit and principles in the present invention, is equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (4)
1. the application of Yi Zhong azolactone chemical compound in the preparation antiradiation drug, Suo Shu azolactone structural general formula is:
X is O in the formula;
Y is the organic group shown in the following formula in the formula:
R wherein
1-R
5, R
7Relatively independent ground or be carbochain, the O (CH of H, 1～20 carbon simultaneously
2CH
2O)
nCH
3(n=0) carbon oxygen chain, Cl, Br, F, carboxylate R
8COO; R
8It is the carbochain of 1～20 carbon; It is characterized in that the application of the people's lung embryo fibroblast MRC-5 damage that X ray or carbon ion beam is caused in treatment.
2. as the application of claim 1 Suo Shu De azolactone chemical compound in the preparation antiradiation drug, it is characterized in that X ray or carbon ion beam irradiation descendant lung embryo fibroblast MRC-5 free radical are had the removing effect.
As claim 1 Suo Shu De azolactone chemical compound in the application of preparation in the antiradiation drug, it is characterized in that the cellular genome damage that X ray or carbon ion beam ionizing radiation are caused has protective action.
As claim 1 Suo Shu De azolactone chemical compound in the application of preparation in the antiradiation drug, it is characterized in that people's lung embryo fibroblast MRC-5 damage that ultraviolet or microwave unionized irradiation are caused has protective action.
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