CN101352438A - Use of deferiprone and its preparations in the preparation of drugs for preventing and treating cardiotoxicity of anthracyclines - Google Patents

Abstract

The invention belongs to the field of medicine and relates to a new medicinal application of deferiprone and its preparation. It specifically relates to the application of deferiprone (1,2-dimethyl-3-hydroxy-4-pyridone, Deferiprone) and its preparations in the prevention and treatment of anthracycline cardiotoxicity. The present invention studies the prevention and treatment effect of deferiprone on the cardiotoxicity of rats caused by doxorubicin from the cell level, tissue level and overall level; Ketone tablets, capsules, oral solutions, and injections. The experimental results confirm that deferiprone and its preparations can not only improve myocardial contractile function and protect the structure of myocardial organelles, but also directly complex free iron ions, significantly reduce the cardiotoxicity and other toxic side effects caused by anthracyclines, and reduce the anthracycline drug-induced mortality.

Description

Use of deferiprone and its preparations in the preparation of drugs for preventing and treating cardiotoxicity of anthracyclines

technical field

The invention belongs to the field of medicine and relates to a new medicinal application of deferiprone and its preparation. It specifically relates to the application of deferiprone (Deferiprone) and its preparations in preventing and treating cardiotoxicity of anthracyclines.

Background technique

Anthracyclines play an important role in the chemotherapy of tumors and have the advantages of broad spectrum and high efficiency. Their main mechanism of action is to directly intercalate between DNA base pairs, interfere with the transcription process, prevent the formation of mRNA, and inhibit the proliferation of tumor cells. . It inhibits both DNA synthesis and RNA synthesis, so it has effects on all stages of the cell cycle, and is a non-specific drug for the cell cycle. However, the dose-cumulative cardiotoxicity of anthracyclines limits their clinical application and increases the pain and burden of patients. Taking Doxorubicin (DOX) as an example, the mortality rate of doxorubicin cardiomyopathy is 40%, and the mortality rate of doxorubicin-induced congestive heart failure is as high as 33%-70%.

One of the ways to prevent and treat cardiotoxicity of anthracyclines is to find cardioprotective agents based on the cardiotoxicity mechanism of anthracyclines. Existing studies have used several drugs to prevent the cardiotoxicity of anthracyclines, and few of them have confirmed efficacy. Currently, only Dexrazoxane has been approved by the US Food and Drug Administration (FDA) for the clinical prevention and treatment of doxorubicin cardiotoxicity. Dexrazoxane has a definite curative effect, but it is expensive, and can only be slowly pushed statically after being dissolved in lactic acid solution. In addition, dexrazoxane can aggravate the bone marrow suppression caused by doxorubicin, and some patients cannot tolerate it. Therefore, it is still necessary to develop a cardioprotective agent with definite curative effect, convenient administration and moderate price.

Deferiprone (1,2-Dimethyl-3-hydroxypyrid-4-one, Deferiprone, DEF) is the first oral iron complexing agent, which was approved by the European Union in 1999 for the treatment of deferoxamine. Contraindicated treatment of iron overload in patients with severe thalassemia due to frequent blood transfusion ^[ Brittenham GM. Iron chelators and irontoxicity [J]. Alcohol, 2003, 30(2): 151-158; and T2 ^* validation and utility[J]. The Lancet, 2003, 361(9352): 182-182; Kontoghiorghes GJ, Agarwal MB, Tondury P, et al.Deferiprone or fatal iron toxic effects? [J]. The Lancet, 2001, 357(9259): 882-883 ^] . Deferiprone belongs to 3-hydroxy-4-pyridone compounds with a molecular weight of 139.2, pKa3.5, and a solubility in water of 16-18 mg/ml at 24°C. At pH 7.4, deferiprone and Fe ³⁺ combine to form a complex (DEF-Fe ³⁺ ) at a ratio of 3: ¹ . ]. Toxicology Letters, 1995, 80(1-3): 1-18. ^] . After oral administration, deferiprone is rapidly absorbed from the stomach, with a plasma half-life ranging from 0.8 hours to 2.3 hours. It is mainly metabolized by the liver and excreted in the urine in the form of gluconic acid or iron complexes. Some studies have used magnetic resonance T2 imaging technology to monitor the myocardial iron content of thalassemia patients who have been taking deferiprone or deferoxamine for a long time. It was found that deferiprone is more effective in reducing myocardial iron content than deferoxamine. The heart function of the treated patients was better, and their cardiac ejection fraction (LVEF) was higher than that of deferoxamine treated patients ^[ Anderson LJ, Wonke B, Prescott E, et al. Comparison of effects of oral deferiprone and subcutaneous desferrioxamine on myocardial iron concentrations and ventricular function in beta-thalassaemia [J]. Lancet, 2002, 360 (9332): 516-520 ^] . In vitro cell test results show that both deferiprone and its Fe ³⁺ complex can effectively prevent ischemia-reperfusion damage to liver cells ^[ Moridani MY, O'Brien PJ.Iron complexes of deferiprone and dietaryplant catechols as cytoprotective superoxide radical scavengers[J].Biochemical Pharmacology, 2001,62(12):1579-1585 ^] , and can reduce the damage of doxorubicin to isolated cardiomyocytes ^[ Link G, Tirosh R, Pinson A, et al.Roleof iron in the Potentiation of anthracycline cardiotoxicity: Identification of heart cell mitochondria as a majorsite of iron-anthracycline interaction [J]. Journal of Laboratory and Clinical Medicine, 1996, 127(3): 272-278;

Barnabe N, Zastre JA, Venkataram S, et al.Deferiprone protects against doxorubicin-induced myocytetotoxicity[J].Free Radical Biology and Medicine,2002,33(2):266-275 ^] . However, there has been no report about the effect of deferiprone on myocardial contractility damage induced by doxorubicin in vitro and the effect on cardiotoxicity of doxorubicin in vivo; Report on formulation development.

Contents of the invention

The purpose of the present invention is to provide new medicinal uses of deferiprone and its preparations. It specifically relates to the application of deferiprone (Deferiprone) and its preparations in preventing and treating cardiotoxicity of anthracyclines.

The present invention first studies the prevention and treatment effect of deferiprone on the cardiotoxicity of rats induced by doxorubicin from the cell level, tissue level and overall level; secondly, a biodegradable deferiprone that can release the drug continuously for 1 month is prepared. Implants, to evaluate the preventive effect of deferiprone on adriamycin cardiotoxicity in rats again; finally, prepare deferiprone tablets, capsules, oral solutions and injections.

The English name of deferiprone described in the present invention is Deferiprone, and its chemical name is 1,2-dimethyl-3-hydroxyl-4-pyridone, which has a chemical structure of formula (I):

Anthracyclines described in the present invention include adriamycin, epirubicin, demethoxydaunorubicin (idarubicin), mitoxantrone (metoxantrone), daunorubicin benzo (rubida-zone) , demethyl daunorubicin (carminomycin), fluoxetine doxorubicin (AD-32), aclacinomycin A (aclacinomycin A) or triiron doxorubicin (quclamycin). Especially doxorubicin.

The cardiotoxicity caused by anthracyclines includes anthracycline-induced cardiomyopathy and chronic cardiac insufficiency.

The dosage range of the deferiprone is 1-20 mg/kg body weight/day, preferably 2.5 mg/kg body weight/day.

The preparation is tablet, capsule, oral solution, injection or implant, preferably tablet, capsule or injection, most preferably tablet or capsule.

Experimental results show that the present invention uses deferiprone (Deferiprone) and its preparations to prevent and treat cardiotoxicity caused by anthracyclines, and the prepared preparations can not only improve myocardial contractility, protect the structure of myocardial organelles, but also directly complex free iron Ions, scavenging oxygen free radicals, reducing myocardial peroxidative damage caused by anthracyclines, reducing the cardiotoxicity of anthracyclines to the human body during chemotherapy, thereby improving the therapeutic index of anthracyclines, reducing the pain and reducing the pain of chemotherapy patients Mortality due to anthracyclines.

Description of drawings

Figure 1 Effect of deferiprone on body weight of doxorubicin rats with heart failure (n=12 ^* )

Among them, ¹ p<0.01, ³ p<0.05 compared with blank control group; ² p<0.01, ⁴ p<0.05 compared with adriamycin group,

Body weight: body weight; Week of experiment: experiment time (week);

CONT: blank control group, DOX: doxorubicin group, DEF-1: deferiprone 1 group,

DEF-2: deferiprone 2 group, DEF-3: deferiprone 3 group, DEF-CONT: deferiprone control group.

The influence of deferiprone on the echocardiographic parameters of rats administered with doxorubicin in the 4th week of Fig. 2 test (n=6),

Among them, Chang: percentage change; EF: ejection fraction, LVFS: left ventricular ejection fraction,

LVEDD: left ventricular internal diameter at end diastole, LVESD: left ventricular internal diameter at end systole,

EDAWT: end-diastolic left ventricular anterior wall thickness, ESAWT: end-systolic left ventricular anterior wall thickness,

EDPWT: end-diastolic left ventricular posterior wall thickness, ESPWT: end-systolic left ventricular posterior wall thickness;

CONT: blank control group, DOX: doxorubicin group, DEF-1: deferiprone 1 group,

DEF-2: deferiprone 2 group, DEF-3: deferiprone 3 group, DEF-CONT: deferiprone control group.

The influence of deferiprone on the echocardiographic parameters of rats with adriamycin heart failure in the 9th week of Fig. 3 test (n=6 ^* ),

Among them, ¹ p<0.01, ³ p<0.05 compared with blank control group; ² p<0.01, ⁴ p<0.05 compared with adriamycin group.

Chang: percent change; EF: ejection fraction, LVFS: left ventricular ejection fraction,

LVEDD: left ventricular internal diameter at end diastole, LVESD: left ventricular internal diameter at end systole,

EDAWT: end-diastolic left ventricular anterior wall thickness, ESAWT: end-systolic left ventricular anterior wall thickness,

EDPWT: end-diastolic left ventricular posterior wall thickness, ESPWT: end-systolic left ventricular posterior wall thickness;

CONT: blank control group, DOX: doxorubicin group, DEF-1: deferiprone 1 group,

DEF-2: deferiprone 2 group, DEF-3: deferiprone 3 group, DEF-CONT: deferiprone control group.

The 8th, 10th week deferiprone of Fig. 4 test is to the influence (n=6) of heart failure rat serum cardiac troponin T of adriamycin,

Among them, ⁴ p<0.05 compared with the Adriamycin group,

cTnT level: cardiac troponin level; Week 8: eighth week, Week 10: tenth week,

DOX: doxorubicin group, DEF-1: deferiprone group 1, DEF-2: deferiprone group 2, DEF-3: deferiprone group 3.

Fig. 5 test the 8th, the 10th week deferiprone is to the influence (n=6) of adriamycin heart failure rat serum urea nitrogen, creatinine,

Among them, ³ p<0.05 compared with the blank control group; ⁴ p<0.05 compared with the doxorubicin group,

BUN: blood urea nitrogen, CRE: creatinine; Week 8: eighth week, Week 10: tenth week,

CONT: blank control group, DOX: doxorubicin group, DEF-1: deferiprone 1 group,

DEF-2: deferiprone 2 group, DEF-3: deferiprone 3 group, DEF-CONT: deferiprone control group.

Figure 6 The effect of deferiprone on the ultrastructure of left ventricular cardiomyocytes in rats with adriamycin heart failure,

Among them, (A): blank control group; (B): doxorubicin group; (C): deferiprone control group; (D): deferiprone 2 groups (A, B, C, D×6000).

Figure 7 The effect of deferiprone implant on the body weight of doxorubicin-administered rats (n=6 ^* ),

Among them, Body weight: body weight,

Time after the first DOX injection (Day): the time (day) after the first administration of doxorubicin,

CONT: blank control group, DEF-I: deferiprone group,

DOX-I: doxorubicin group, DOX+DEF-I: doxorubicin+deferiprone group.

The effect of Fig. 8 deferiprone implant on myocardial tissue structure of Adriamycin-administered rats,

Among them, (A): blank control group; (B): doxorubicin group; (C): doxorubicin+deferiprone group (A, B, C×100).

Fig. 9 In vitro dissolution of common deferiprone tablets (n=6).

Figure 10 In vitro release of deferiprone sustained-release tablets (n=6).

Fig. 11 The average blood drug concentration-time curves (n=6) after a single oral dose of deferiprone common tablet and sustained-release tablet.

Detailed ways

Example 1 Deferiprone (Deferiprone) prevents and treats cardiotoxicity in rats caused by doxorubicin (DOX)

1.1 Establishment of doxorubicin heart failure model and dose setting of deferiprone The experiment lasted for 10 weeks, the drug was stopped at the 8th week, and the observation continued for 2 weeks. Get 72 rats, divide into 6 groups at random according to body weight, administer according to the following method:

Adriamycin group: intraperitoneal injection of 2.5 mg/kg (4.31 μmol/kg) of adriamycin, once a week, for 8 consecutive weeks.

Blank control group: intraperitoneal injection of equal volume of normal saline, 5 days a week, for 8 consecutive weeks.

Deferiprone treatment group: Different doses of deferiprone were administered 5 days a week for 8 consecutive weeks. in:

Deferiprone group 1: the treatment was the same as that of the doxorubicin group, and a low dose of deferiprone 1.71 mg/kg was given intraperitoneally.

Deferiprone group 2: the treatment was the same as that of the doxorubicin group, and a middle dose of deferiprone 6.85 mg/kg was given intraperitoneally.

Group 3 of deferiprone: the treatment was the same as that of the doxorubicin group, and a high dose of deferiprone (13.71 mg/kg) was given intraperitoneally.

Deferiprone control group: deferiprone injections were given, and the dosage and administration interval were the same as those of deferiprone 3 groups.

1.2 General toxicity observation

The diet and activity status of the rats were observed daily from week 1 to week 6 of the test, and observed twice a day from week 7 until the end of the test. The body weight of rats in each group was recorded once a week. Dead rats were weighed and dissected to measure ascites volume. In the 10th week of the experiment, the rats in each group were anesthetized with pentobarbital sodium (36 mg/kg), and the heart was quickly removed by thoracotomy, rinsed with cold saline, fixed for electron microscope observation, and the volume of ascites was measured at the same time.

1.2.1 Weight change

The body weights of the animals in each group before the test are shown in Table 1, and the body weight changes of the animals in each group during the test are shown in Figure 1.

After injecting doxorubicin, the weight gain of the doxorubicin group and the deferiprone treatment group slowed down to varying degrees. At the 4th week of the test (accumulative dose of doxorubicin 7.5mg/kg), the weight gain of the doxorubicin group and the deferiprone treatment group began to be significantly lower than that of the blank control group and the deferiprone control group (p<0.05); At 9 weeks, that is, 1 week after doxorubicin was discontinued (the cumulative dose of doxorubicin reached 20 mg/kg), the body weight of the deferiprone 2 and 3 groups began to be greater than that of the doxorubicin group (p<0.05); at the 10th week, That is, 2 weeks after doxorubicin withdrawal, the deferiprone 1 group weighed more than the doxorubicin group (p<0.05).

Rats in the deferiprone control group had no obvious adverse reactions, and their weight gain was similar to that of the blank control group.

Table 1 shows the body weights of the animals in each group (n=12) before the test, and the body weight changes of the animals in each group during the test are shown in FIG. 1 .

Table 1

1.2.2 Symptoms and signs and ascites volume

The doxorubicin group appeared listless, curled up and shrugged, and short of breath at the 7th week of the test; at the 9th week of the test (the cumulative dose of doxorubicin reached 20mg/kg), they were obviously thin; at the end of the test, obvious edema of the liver and kidney was seen That is, a large amount of ascites. The deferiprone treatment group had milder symptoms, and the amount of ascites in the deferiprone 1 and 2 groups was significantly lower than that in the doxorubicin group (p<0.01 or 0.05). Rats in the blank control group and the deferiprone control group had no obvious adverse reactions, and there were no liver and kidney edema and ascites at the end of the test. Table 2 shows the amount of ascites and the survival rate of rats in each group in the 10th week of the experiment.

Table 2

² p<0.01, ⁴ p<0.05 compared with the doxorubicin group.

1.2.3 Animal survival rate

No animal died in the blank control group, deferiprone control group, and deferiprone 2 groups, and the survival rate was 100%. 3 rats in the doxorubicin group died at the 8th, 9th and 10th week of the experiment, and the survival rate was 75%; 1 rat in the deferiprone 1 and 3 groups died at the 10th week, and the survival rate was 75%. 91.7% (Table 2).

1.3 Echocardiographic monitoring

The cardiac function of the rats in each group was measured 3 days before the test and at the 4th week (the cumulative dose of adriamycin reached 7.5 mg/kg) and the 9th week (the cumulative dose of adriamycin reached 20 mg/kg) of the test.

Image acquisition: Rats were anesthetized ip with pentobarbital sodium (36mg/kg), fixed in the supine position, and had their chest hair removed. M-mode ultrasonography (Philips Sonos 5500 multifunctional ultrasonic diagnostic instrument, S12 probe, speed 100mm/s) was performed at the largest lumen diameter of the papillary muscle in the short axis view of the parasternal left ventricle, and the left ventricle diameter at the end of diastole and end of systole was measured ( LVEDD and LVESD), end-diastolic and end-systolic left ventricular anterior wall thickness (EDAWT and ESAWT), end-diastolic and end-systolic left ventricular posterior wall thickness (EDPWT and ESPWT). The instrument automatically calculates left ventricular shortening fraction LVFS(%)=(LVEDD-LVESD)×100/LVEDD, and left ventricular ejection fraction (LVEF, Teichholz method) ^[ Bertinchant JP, Polge A, Juan JM, et al.[J] . Clinica Chimica Acta, 2003, 329: 39-51; Huang Guoqian, Liu Hong, Shu Xianhong, et al. [J]. Chinese Journal of Ultrasound Imaging, 2004, 13: 848-852 ^] . All data were videotaped and analyzed off-line, and counting data were continuously measured for 3 cardiac cycles to obtain the average value.

Based on the echocardiographic parameters (Index ₀ ) before the test, the percentage change (%) of the data (Index') in the 4th and 9th weeks was calculated according to the following formula:

$\mathrm{<span\; class="notranslate">\; change</span>}<span\; class="notranslate">\; (</span><span\; class="notranslate">\; \%</span><span\; class="notranslate">\; )</span><span\; class="notranslate">\; =</span><span\; class="notranslate">\; (</span>\underset{<span\; class="notranslate">\; \&OverBar;</span>}{\frac{\mathrm{<span\; class="notranslate">\; Inde</span>}{\mathrm{<span\; class="notranslate">\; x</span>}}^{<span\; class="notranslate">\; ,</span>}}{{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="0"\; label="Index"\; filenames="A20071004420700162.png,A20071004420700171.png"\; state="\{\{state\}\}">Index</figure-callout></span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}}}<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; \&times;</span>\mathrm{<span\; class="notranslate">\; 100</span>}<span\; class="notranslate">\; \%</span>$

The results showed that there was no significant difference in the echocardiographic indexes of the rats in each group before the test and at the 4th week of the test (the cumulative dose of adriamycin reached 7.5 mg/kg) (Fig. 2).

At the 9th week of the test, compared with the blank control group, the left ventricular shortening fraction (LVEF) and left ventricular ejection fraction (LVFS) in the adriamycin group were significantly reduced (p<0.01), while the left ventricular diameter at the end of diastole and end of systole ( LVEDD and LVESD) were significantly increased (p<0.01), and pericardial effusion was present in 2 animals (Fig. 3).

Cardiac function was maintained to varying degrees in each deferiprone treatment group, and no pericardial effusion was detected in the 9th week of the trial. Compared with the doxorubicin group, the left ventricular shortening fraction (LVEF) and left ventricular ejection fraction (LVFS) in the deferiprone 2 group were significantly increased, and the LVESD was significantly decreased (p<0.01); The inner diameter (LVESD) decreased significantly (p<0.01); the left ventricular ejection fraction (LVFS) in the deferiprone 3 group increased significantly (p<0.05). The deferiprone control group had no significant difference with the blank control group in all indexes, and there were significant differences compared with the doxorubicin group (p<0.01).

1.4 Monitoring of serum biochemical indicators

Blood was collected 2 hours after intraperitoneal injection of doxorubicin in the 8th week of the test and at the 10th week of the test, and the serum was separated to detect cardiac troponin T (cTnT), blood urea nitrogen (BUN), creatinine (CRE) and triglyceride (TG) and other indicators.

Cardiac troponin T detection: the electrochemiluminescence method was used, and the detection equipment was ELECSYS-2010 automatic electrochemiluminescence immunoassay analyzer (Roche, Japan), and the reagents were all products of Roche Company. Reference value <0.1ng/ml.

Detection of blood urea nitrogen, creatinine, and triglycerides: detected by VITROS-950 automatic biochemical analyzer (Johnson & Johnson, USA), and the reagents are all products of Johnson & Johnson.

1.4.1 Monitoring results of serum cardiac troponin T

No cardiac troponin T was detected in the serum of the blank control group and the deferiprone control group at the 8th week and the 10th week (detection limit: 10 ng/L). Serum cardiac troponin T in the doxorubicin group increased at week 8, and increased to 11.31 times that at week 8 at week 10, p<0.05 (Figure 4).

Serum cardiac troponin T in the deferiprone treatment group at week 10 was higher than that at week 8, but there was no significant difference. Compared with the doxorubicin group, the serum cardiac troponin T in the deferiprone 2 and deferiprone 3 groups decreased significantly at week 10 (p<0.05).

1.4.2 Monitoring results of serum urea nitrogen and creatinine

2 hours after doxorubicin injection in the eighth week, there was no significant difference in serum urea nitrogen and creatinine of rats in each group (Fig. 5).

At week 10, blood urea nitrogen and creatinine did not increase significantly in the blank control group and deferiprone control group. Compared with the blank control group, the blood urea nitrogen and creatinine in the adriamycin group were significantly increased (p<0.05). Compared with the doxorubicin group, the blood urea nitrogen and creatinine in the deferiprone 2 group and deferiprone 3 group decreased significantly at week 10 (p<0.05).

1.5 Myocardial Ultrastructural Observation

Myocardial ultrastructure was observed by transmission electron microscope (Fig. 6). Adriamycin group (B) had myocardial cell edema, mitochondrial swelling, sarcoplasmic reticulum expansion, and loose arrangement of myofibrils. In contrast, in the deferiprone 2 group (D), cardiomyocyte damage was generally mild, with little mitochondria swelling, myocardial fibers arranged neatly, and only a few sarcoplasmic reticulum slightly expanded. There was no obvious myocardial lesion in the blank control group (A) and the deferiprone control group (C).

The above experimental results of the present invention show that deferiprone can reduce the damage of doxorubicin to cardiac contractile function and damage to myocardial cell organelles in vivo, and can prevent persistent myocardial damage caused by doxorubicin.

Example 2

Accurately weigh deferiprone, lactic acid-glycolic acid copolymer (PLGA) and/or polylactic acid (PLA) according to a certain proportion, dissolve them in an appropriate amount of dichloromethane, put them in an oil bath at 37°C, and evaporate the solvent to make a self-made extruder Extrusion. After drying in a vacuum oven at room temperature to constant weight, it was cut into small rods with a diameter×length of 2mm×20mm, and the average weight of the preparation was 84.23±11.01mg to obtain the implant of the present invention.

Example 3

Prevention and treatment of deferiprone biodegradable implants on cardiotoxicity induced by doxorubicin in rats

3.1 Grouping and administration

The experiment was divided into 4 groups and lasted 30 days. Table 3 is the dosage regimen of each group.

table 3

After administration, the general situation was observed, and the changes in body weight and survival rate were recorded. Animals that died during the experiment were weighed and dissected in time. The experiment was ended on the 30th day after administration, and the ascites volume and liver and kidney edema were recorded after the animals were sacrificed by bloodletting; the left ventricular muscle at the apex of the heart was fixed with 10% formaldehyde for pathological examination.

3.2 Effect of deferiprone implant on the survival rate of doxorubicin-administered rats

After the implantation operation, there was no redness, swelling and exudation in the wound on the back of the rat, and it was basically healed 1 week after the operation, and the wound healed well. The results of the survival rate of rats are shown in Table 4, which shows the effect of deferiprone implants on the survival rate of rats administered with doxorubicin.

Table 4

Animals died in both the doxorubicin group and the doxorubicin+deferiprone group. Animals in the doxorubicin group died on the 1st to 18th day after the first dose of doxorubicin, and only 2 animals remained after 30 days of administration, with a survival rate of 33.33%. Animals in the doxorubicin+deferiprone group died on the 1st to 12th day after the first administration of doxorubicin, and the survival rate after 30 days of administration was 66.67%. The other two groups of animals all survived to the end of the experiment.

3.3 Effects of deferiprone implants on the body weight of doxorubicin-administered rats

After the preparation was implanted, the patients were weighed twice a week. The results showed that during the test period, the body weight of the blank control group and the deferiprone group increased with time, and there was no difference between the groups. The body weights of the animals in the other two groups all decreased significantly. On the 4th day of the test (the cumulative dose of adriamycin reached 2.5mg/kg), the body weight of the adriamycin group and the adriamycin+deferiprone group began to be lower than that of the blank control group (p<0.05); After 4 days of drug withdrawal, the cumulative dose reached 10 mg/kg), the body weight of the doxorubicin+deferiprone group was greater than that of the doxorubicin group (p<0.05). (Figure 7)

3.4 Effect of deferiprone implant on ascites in rats administered with doxorubicin

The results showed that after administration of doxorubicin, the rats developed liver and kidney edema combined with severe ascites. In terms of the incidence of ascites, ascites occurred in both the doxorubicin group and the doxorubicin+deferiprone group, accompanied by liver and kidney edema, but the incidence of ascites in the doxorubicin+deferiprone group was lower; In terms of degree, compared with the Adriamycin group, the amount of ascites in the Adriamycin+Deferiprone group was less (p<0.05).

Table 5 shows the effect of deferiprone implants on the ascites of rats administered with doxorubicin (n=6).

table 5

⁴ p<0.01 compared with Adriamycin I group

3.5 Effects of deferiprone implants on myocardial tissue structure in rats administered doxorubicin

The results showed that in the myocardium of rats in the blank control group (A), the morphology and structure of the heart tissue were intact, the layers of cardiomyocytes were clear, and no abnormal pathological changes were seen ( FIG. 8 ).

In the doxorubicin group (B), the cardiomyocytes in the heart tissue of the rats were hypertrophic, the muscle fibers were loose, and vacuolar degeneration was seen in some cardiomyocytes. The degree and extent of cardiac tissue lesions in the doxorubicin+deferiprone group (C) rats were lighter than those in the doxorubicin group, with a small amount of hypertrophy of cardiomyocytes, loose muscle fiber structure, and mild vacuolar degeneration.

It was found that compared with the doxorubicin group, the doxorubicin + deferiprone group had a lower incidence of ascites, a better general condition, and milder myocardial pathological damage, suggesting that deferiprone implants can be used to a certain extent. Relief of doxorubicin-induced cardiotoxicity in rats. The results of this experiment suggest that deferiprone can partially alleviate the cardiotoxicity of doxorubicin in rats at a sustained low blood concentration.

3.6 Effect of deferiprone implant on myocardial iron content of rats administered with doxorubicin

Take about 100 mg of left ventricular muscle at the apex of rats in the blank control group, doxorubicin group, doxorubicin I group, doxorubicin+deferiprone I group, and use the flame method to measure the iron content, which includes free iron content and Non-Free Iron Content. The technical specifications refer to GB/T15337-94 General Rules of Atomic Absorption Spectroscopic Analysis, and the measurement range and accuracy are (190~900)nm±0.5nm. Table 6 shows the effect of DEF implants on myocardial iron content of DOX-administered rats.

Table 6

^* At the end of the experiment, 4 rats in the Adriamycin I group died, and 2 rats in the Adriamycin+Deferiprone I group died

Only 2 rats in the Adriamycin group survived to the end of the experiment, and the myocardial iron content was higher than that of other groups. There was no difference in the myocardial iron content of rats in the blank control group and the doxorubicin+deferiprone group.

Example 4

Take 500mg of deferiprone, dissolve it in 1000ml of water for injection, add 2g of activated carbon, heat at 100°C for 5 minutes, decarburize by coarse filtration, adjust the pH to 7 with sodium hydroxide test solution, and filter until clarified. Potting, and sterilizing at 100°C for 30 minutes, the injection of the present invention is obtained.

Example 5

Take 500 mg of deferiprone, add 200 mg of compressible starch, 200 mg of lactose, and 400 mg of microcrystalline cellulose, mix well, granulate, dry, and granulate to obtain the granule of the present invention.

Example 6

Take 500 mg of deferiprone, add 200 mg of compressible starch, 200 mg of lactose, and 400 mg of microcrystalline cellulose, mix evenly, granulate, dry, granulate, and pack into capsules to obtain the capsule of the present invention.

Example 7

Take 500 mg of deferiprone, add 1300 mg of microcrystalline cellulose and 10 mg of magnesium stearate, mix well, add 60% ethanol containing 6% PVP to moisten, then granulate, dry, granulate, and compress to obtain the present invention. Ordinary tablets (each tablet contains deferiprone 25mg).

The dissolution rate was determined by the basket method specified in the Chinese Pharmacopoeia 2005 edition. Use 900mL of degassed purified water as the release medium, the temperature is (37±0.5)°C, and the rotation speed is 50r/min, sample 5mL at 3, 5, 10, 15, 20, and 25 minutes respectively (at the same time, immediately add water of the same temperature 5 mL), filtered through a 0.8 μm microporous membrane, discarded the initial filtrate, and kept the filtrate for testing. After sampling at each time point, the samples were measured by UV to obtain their in vitro dissolution curves ( FIG. 9 ).

Example 8

Take 500 mg of deferiprone, add 300 mg of hydroxypropyl methylcellulose (HPMC) K100M, 888300 mg of wax, 700 mg of microcrystalline cellulose, and 10 mg of magnesium stearate, mix well, add 60% ethanol containing 6% PVP to moisten After granulation, drying, granulation, and tabletting, the sustained-release tablets of the present invention (each containing 25 mg of deferiprone) are obtained.

The release rate was determined by the basket method stipulated in the Chinese Pharmacopoeia 2005 edition. Use 900mL of degassed purified water as the release medium, the temperature is (37±0.5)°C, and the rotation speed is 50r/min, sample 5mL at 1, 2, 4, 6, 8, 10, and 12 hours respectively (while immediately replenishing the same temperature 5 mL of water), filtered through a 0.8 μm microporous membrane, discarded the initial filtrate, and kept the filtrate for testing. After sampling at each time point, the samples were measured by UV to obtain their in vitro release curve ( FIG. 10 ).

Example 9

Comparison of Pharmacokinetic Behavior of Deferiprone Ordinary Tablets and Sustained-release Tablets in Animals

A two-period two-drug crossover design was adopted. Six adult male Beagle dogs were weighed and randomly divided into two groups, one for the test drug group and the other for the reference drug group, with 3 dogs in each group. After fasting for 12 hours, the test animals were orally administered 25 mg of the test preparation deferiprone sustained-release tablet or 25 mg of the reference preparation deferiprone ordinary tablet. The blood sampling time of the test preparation is 20, 40, 60, 90, 120, 180, 240, 300, 360, 420, 480, 540, 600, 720 min before and after taking the medicine; the blood sampling time of the reference preparation is 10 minutes before and after taking the medicine. , 15, 20, 30, 45, 60, 90, 120, 180, 240, 300, 360, 480, 720min. 3mL of blood was collected from the forelimb vein, bathed in 37°C water for 30min, centrifuged at 3000r/min for 15min, and the supernatant serum was collected and stored at -20°C until assayed. One week after drug withdrawal, a crossover test was carried out in the same way as above.

The average plasma concentration-time curves of deferiprone ordinary tablets and sustained-release tablets after single-dose oral administration of 6 beagle dogs are shown in Figure 11, and the pharmacokinetic parameters were calculated by rectangle method.

Table 7 shows the pharmacokinetic parameters (x±s, n=6) after single-dose oral administration of deferiprone sustained-release tablets and common tablets to beagle dogs.

Table 7

Note: t _1/2 : biological half-life, t _max : peak time, C _max : peak concentration, AUC _0-720 : area under the plasma concentration-time curve from 0 to 720 minutes, AUC _0～∞ : from 0 plasma concentration-time curve from time to infinity

Area under the line, MRT _0-∞ : mean residence time, F%: relative bioavailability.

_The results showed that the absorption degree of deferiprone sustained-release tablets and deferiprone _ordinary tablets were bioequivalent; The peak concentration (C _max ) decreased, indicating that the sustained-release tablet had obvious sustained-release characteristics.

The medicines used in the experiment of the present invention are commercially available, and the experimental animals used are purchased from the former Department of Experimental Animals of Shanghai Medical University.

Claims (8)

1. The deferiprone with the structure of formula (I) and the application of the preparation thereof in preparing the medicine for preventing and treating anthracycline cardiotoxicity,

2. use according to claim 1, characterized in that the anthracycline is selected from the group consisting of doxorubicin, epirubicin, demethoxydaunorubicin, mitoxantrone, daunorubicin, nordaunorubicin, fluoroethylene doxorubicin, aclarubicin A, and triiron doxorubicin.

3. The use according to claim 1, characterized in that said anthracycline is doxorubicin.

4. The use according to claim 1, characterized in that the anthracycline cardiotoxicity is anthracycline cardiomyopathy or anthracycline chronic cardiac insufficiency.

5. The use according to claim 1, wherein the dosage of the ferrone is 1-20 mg/kg body weight/day.

6. Use according to claim 5, characterized in that the dosage of said ferrone is 2.5mg/kg body weight/day.

7. Use according to claim 1, characterized in that the preparation is selected from the group consisting of tablets, capsules, oral solutions, injections or implants.

8. Use according to claim 7, characterized in that the formulation is selected from tablets or capsules.

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