Low-hepatotoxicity actinomycin D solid dispersion and preparation method thereof

Abstract

The invention relates to a low-hepatotoxicity actinomycin D solid dispersion and a preparation method thereof. Actinomycin D, a plant source functional natural auxiliary material, a flow aid and ball-milling beads are placed in a ball-milling tank for co-grinding reaction to obtain the low-hepatotoxicity actinomycin D solid dispersion product. The solid dispersion is prepared through mechanochemical grinding, no organic solvent is needed, degradation of drugs in the high-temperature melting process is avoided, the drugs and auxiliary materials are easy to amorphize, the solubility and bioavailability of the drugs are improved, and the solid dispersion has the advantages of being high in preparation efficiency, low in production cost, easy to operate, environmentally friendly and the like; and plant-derived macromolecules are adopted as carriers, so that the hepatotoxicity of actinomycin D is reduced while solubilization and synergism are realized, and the composition is crucial to safe medication of antibiotic drugs and also has positive significance in exploring natural, non-toxic, efficient and multifunctional drug-loading system auxiliary materials.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical preparations, and particularly relates to a low hepatotoxicity actinomycin D solid dispersion and a preparation method thereof. The solid dispersion prepared by the defined method can improve the bioavailability and reduce hepatotoxicity. BACKGROUND

[0002] Actinomycin D (ACD) is a potent polypeptide antitumor antibiotic and a cell cycle non-specific drug, which is one of the most classic anticancer drugs in clinical practice. ACD can insert into the DNA double helix and form a stable complex with guanine residues, block the function of RNA polymerase, and ultimately interfere with protein synthesis, thereby exerting strong antitumor activity. Compared with other alkylating agents or topoisomerase inhibitors, actinomycin D has the characteristics of fast onset, long-lasting effect, and significant effect on various solid tumors (such as nephroblastoma, rhabdomyosarcoma, Ewing's sarcoma, and trophoblastic tumor). However, actinomycin D can also cause severe dose-dependent hepatotoxicity: since the 1960s, cases of fatal liver damage, hepatic veno-occlusive disease (VOD), and acute liver failure have been reported. Therefore, the US FDA has set strict restrictions on the cumulative dose, infusion rate, and combination of actinomycin D, and the European Union also requires the addition of a black box warning in the instructions. Actinomycin D itself has very low water solubility (<0.1 mg / mL), and high-concentration infusion is required to maintain an effective blood drug concentration, which further increases the liver first-pass metabolism burden, leading to hepatocyte necrosis, fatty degeneration, cholestasis, and even progression to liver failure and life-threatening. Therefore, as a classic anticancer drug, actinomycin D has a narrow therapeutic window and is strictly limited in clinical application due to its severe dose-dependent hepatotoxicity and extremely low water solubility. However, as an irreplaceable core antitumor drug in clinical practice, how to maintain the high antitumor activity of actinomycin D while significantly reducing its fatal hepatotoxicity is of great significance to improve the safety of the drug and further improve the safety of actinomycin D.

[0003] In recent years, plant extracts, especially natural high polymers, have been increasingly used as drug carriers. Natural high polymers from plants have the advantages of low cost, high availability, high safety, good biocompatibility and biodegradability, and play an important role in improving drug solubility and stability, achieving drug sustained release, targeted delivery and controlled release, improving drug efficacy and reducing side effects. For example, cellulose, gelatin, xanthan gum, pectin, alginate and other high polymers from plants and animals have been used for drug stabilization, solubilization, microencapsulation and functional formulation development. Some natural high polymers from plants have certain biological activities, for example, astragalus polysaccharide and glycyrrhizin polysaccharide have immunomodulatory, anti-inflammatory, anti-tumor and liver protection effects, and poria cocos polysaccharide is reported to have liver protection activity by reducing oxidative stress and inflammation, but the mechanism has not been elucidated. At the same time, plant polysaccharides are rich in polar groups such as hydroxyl and carboxyl groups, and have regular spatial structures, and a large number of hydroxyl groups between molecules can encapsulate non-water-soluble molecules into small particles, which have the potential to be used as drug carriers. Therefore, using plant-derived polysaccharides with special physiological activity as the main carrier and using green and environmentally friendly mechanical-chemical solvent-free preparation technology to prepare amorphous actinomycin D solid dispersion can achieve the dual effects of reducing liver toxicity and increasing drug efficacy of actinomycin D, which has a positive significance for exploring natural, non-toxic and high-efficiency multifunctional drug delivery system adjuvants, and provides a new idea for fully utilizing Chinese herbal medicine resources and developing integrated traditional Chinese and Western medicine research. SUMMARY

[0004] In view of the problems of low bioavailability and liver toxicity of existing antibiotic drug actinomycin D preparations, the purpose of the present application is to provide a low liver toxicity actinomycin D solid dispersion and a preparation method thereof. The present application uses plant-derived functional natural product adjuvants with clear liver protection activity as drug carriers, and uses solvent-free mechanical ball milling technology to construct a new drug delivery system with "treatment-liver protection" integration. This method can increase solubility and efficacy, solve the problem of fatal liver toxicity in the clinical application of actinomycin D, significantly improve its safety, and also improve the bioavailability, and the production process is green and environmentally friendly.

[0005] The present application defines a low liver toxicity actinomycin D solid dispersion, which uses plant-derived functional natural products as drug carriers and actinomycin D as drugs. The plant-derived functional natural products are physically dispersed with the drug actinomycin D under solvent-free conditions by mechanical grinding method to form a uniform solid dispersion, and biologically resist liver damage caused by the drug.

[0006] Further, the plant-derived functional natural product adjuvant is selected from one of poria cocos polysaccharide, polygonatum polysaccharide, mannitol, glycyrrhizic acid or glycyrrhizic acid disodium, and preferably poria cocos polysaccharide.

[0007] Further, the application also limits the preparation method of the low hepatotoxicity actinomycin D solid dispersion, specifically: the drug actinomycin D, the drug carrier plant-derived functional natural product, the flow aid and the ball milling beads are placed in a ball milling jar for co-grinding reaction, and the grinding is carried out at a rotating speed of 100-400 rpm for 0.5-4 h, so that the drug actinomycin D is uniformly dispersed in the drug carrier plant-derived functional natural product, and the low hepatotoxicity actinomycin D solid dispersion is obtained.

[0008] Further, the application also limits the flow aid to be one of micro-powder silica gel, magnesium stearate or talc powder.

[0009] Further, the application also limits the mass ratio of the actinomycin D and the plant-derived functional natural product material to be 1:1-20, preferably 1:7.

[0010] Further, the application also limits the mass ratio of the actinomycin D and the flow aid to be 20-100:1, preferably 50:1.

[0011] Further, the application also limits the diameter of the ball milling beads to be 5-10 mm, preferably 7.5 mm, and the number of the ball milling beads to be 5-10, i.e. the mass ratio of the ball and the material is 45:1-90:1, preferably 72:1. (In the embodiment of the application, the total amount of the feeding material is limited to 802 mg, the volume of the ball milling jar is 50 mL, the diameter of the ball milling bead is 7.5 mm, and the weight of each steel bead is about 7.2 g) Further, the application also limits the ball milling instrument to be a planetary ball mill, the rotating speed is 100-400 rpm, and the grinding time is 2 h.

[0012] By using the above-mentioned technology, the application has the following beneficial effects compared with the prior art: 1) The core of the application is that, by innovative preparation technology and ingenious carrier selection, a key clinical problem of a classic anticancer drug (actinomycin D), i.e. severe hepatotoxicity, is solved, and the dual effects of treatment and liver protection are realized by "drug assisted by drug", i.e. a natural polysaccharide (such as krestin) with definite liver protection activity is selected as the main carrier, which surpasses the role of traditional auxiliary materials as inert carriers to improve the solubility, the carrier not only disperses the drug and solubilizes in a physical way, but also actively neutralizes the liver damage caused by the drug by using the pharmacological action of the carrier itself, so that the synergy of "1 (drug) + 1 (auxiliary material) > 2" is realized, the bioavailability of the antitumor drug is improved, and the liver toxicity caused by the drug is reduced by the carrier itself; 2) The preparation process of the present application discards a large amount of organic solvents, which has problems such as solvent residue, environmental pollution, complex process and risk of drug degradation, etc. The mechanical force chemical ball milling method is used to form a uniform solid dispersion of the drug and the excipient under solvent-free conditions through physical grinding. This method is green, simple, safe and reliable, and conforms to the development trend of modern pharmaceutical industry; 3) The solid dispersion obtained by the present application can greatly improve the solubility and bioavailability of actinomycin D, effectively reduce its fatal hepatotoxicity by using the liver-protecting effect of the active carrier, and widen the therapeutic window, thereby significantly improving the safety. The improvement of bioavailability of the present application means that the required dose to achieve the same efficacy can be reduced, further reducing the risk of toxic side effects, achieving the clinical goal of "attenuation and efficiency", thereby increasing the efficacy, and the operation is simple, avoiding the use of organic solvents, preventing problems such as raw material loss and thermal decomposition caused by solvent residue and solvent thermal removal process, and having the advantages of high preparation efficiency, safe and reliable production, low production cost, less pollution, etc. It is easier to industrialize, opens up a new way for the application of active ingredients (polysaccharides) in traditional Chinese medicine in modern high-value drug preparations, and promotes the in-depth development of the combination of traditional Chinese and Western medicine. BRIEF DESCRIPTION OF DRAWINGS

[0013] Figure 1 is a dissolution curve of actinomycin D in a medium with pH = 1.2; Figure 2 is a dissolution curve of actinomycin D in a medium with pH = 6.8; Figure 3 is an in-vitro membrane permeation curve of actinomycin D; Figure 4 is an X-ray diffraction spectrum of actinomycin D, Example 1 and Example 2; Figure 5 is a differential scanning calorimetry spectrum of actinomycin D, Example 1 and Example 2; Figure 6 is a curve of drug content change in the plasma of rats after gavage with different actinomycin D; Figure 7 is a graph of serum ALT content of rats after gavage with different actinomycin D; Figure 8 is a graph of serum AST content of rats after gavage with different actinomycin D; Figure 9 is a graph of serum ALP content of rats after gavage with different actinomycin D. DETAILED DESCRIPTION

[0014] The present application will be further described below in conjunction with specific embodiments, but the protection scope of the present application is not limited thereto.

[0015] Example 1 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0016] Example 2 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, rhizoma polygonati polysaccharide 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0017] Example 3 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, mannitol 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0018] Example 4 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, glycyrrhizic acid 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0019] Example 5 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, disodium glycyrrhizinate 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0020] Example 6 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, krestin 700 mg, magnesium stearate 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0021] Example 7 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, krestin 700 mg, talc 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0022] Example 8 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 300 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0023] Example 9 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 1000 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0024] Example 10 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 1500 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0025] Example 11 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 1 mg, and stainless steel ball milling beads 8 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0026] Example 12 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 5 mg, and stainless steel ball milling beads 8 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0027] Example 13 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 5 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0028] Example 14 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 10 pieces of 7.5 mm in diameter, sequentially, and mill for 2 h at 200 rpm. After the ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0029] Example 15 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 0.5 h. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0030] Example 16 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 1 h. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0031] Example 17 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 200 rpm for 4 h. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0032] Example 18 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 10 pieces with a diameter of 7.5 mm in sequence, and mill at 100 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0033] Example 19 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 300 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0034] Example 20 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel ball milling beads 8 pieces with a diameter of 7.5 mm in sequence, and mill at 400 rpm for 2 h. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0035] Example 21 Actinomycin D standard curve drawing: Precisely take 20 mg of actinomycin D standard (source leaf, 98%) and place it in a 50 mL volumetric flask. Add the mobile phase to dissolve, dilute to the mark, sonicate to dissolve, and shake to mix. The resulting solution is used as the stock solution. Precisely take 0.1, 0.5, 1, 2.5, 3, 3.5, 4, 5, 6 mL of the stock solution and dilute to the mark with the mobile phase in 9 10 mL volumetric flasks. Shake to mix. Dilute to 4, 20, 40, 100, 120, 140, 160, 200, 240 ug / mL, respectively. Filter through a 0.22 μm filter membrane. Determine the content by HPLC. The HPLC conditions are as follows: detector: UV / VIS detector; column: C18 column (4.6 x 150 mm, 5 μm); column temperature: 25 °C; flow rate: 1.0 mL / min; injection volume: 10 μL; mobile phase: methanol: acetonitrile: water (65:10:25); detection wavelength: 254 nm.

[0036] Obtain the peak areas corresponding to different concentrations of actinomycin D. Perform linear regression of the peak areas versus the concentrations. The regression equation is y = 19891x - 12793 (R 2 = 0.9999) with a concentration range of 4 μg / mL to 250 μg / mL.

[0037] Dilute the solution to be determined to a peak area within the range of the standard curve. Record the dilution factor as V. Determine the peak area PA of the solution by HPLC. Calculate the concentration c (ug / mL) of actinomycin D in the solution to be determined according to the following formula.

[0038] Formula 1: c (ug x mL -1 ) = V x (PA + 2793) ÷ 19891.

[0039] Example 22: Determination of the solubility and preparation yield of actinomycin D prepared by the examples The method is as follows: take an excess amount of the prepared actinomycin D solid dispersion, place it in a conical flask, add 20 mL of distilled water, and place it in a shaking bed to reach the saturated concentration. The shaking bed parameters are a temperature of 37 °C, a rotation speed of 200 rpm, and a time of 24 h. Filter the saturated solution using a 0.22 μm filter membrane and determine the peak area PA by HPLC. Calculate the concentration of actinomycin D in the saturated solution of the complex in water according to the standard curve described in Example 21. The solubility of the actinomycin D solid dispersions described in Examples 1-20 is shown in Table 1. At the same time, the selection and effect of the flow aid are investigated by the preparation yield of actinomycin D. The preparation yield = actual product amount / theoretical product amount x 100%. The equilibrium solubility and yield of each example are shown in Table 1. The results show that the solubilizing effect of the actinomycin D solid dispersions in Examples 1 and 2 is better, the powder flowability is good, and the preparation yield is high. Examples 1 and 2 are selected for further physicochemical property and pharmacodynamic characterization.

[0040] Table 1 Formulation solubility and preparation yield parameters .

[0041] Example 23 Dissolution of prepared solid dispersion of dactinomycin The in vitro dissolution rates of the samples were tested according to the second method of dissolution test in Chinese Pharmacopoeia 2020 edition (paddle method). The dactinomycin raw material and the drug delivery system of examples 1 and 2 with a theoretical drug content of 50.0 mg were weighed, and potassium dihydrogen phosphate and sodium hydroxide were used to prepare phosphate buffer solutions with pH = 1.2 and pH = 6.8 as dissolution liquids. The temperature of the dissolution tester was set to 37.0 ± 0.5 °C, and the paddle speed was 75 rpm. 5 mL of sample liquid was taken at 5, 10, 15, 20, 30, 40, 60, 90, 120, 150, 180 min, and immediately supplemented with an equal volume of fresh dissolution medium. The sample liquid was filtered through a 0.22 µm water membrane, and the concentration was determined by high performance liquid chromatography using the method described in example 21. The peak area was converted to concentration, and the dissolution curve was plotted using the concentration points. The dissolution curves are shown in Figure 1 and Figure 2 . As shown in Figure 1 and Figure 2 , the release rate and dissolution effect of dactinomycin were significantly improved in both pH = 1.2 and pH = 6.8 dissolution media. The final dissolution of example 1 in pH = 1.2 and pH = 6.8 dissolution media increased by about 38% and 60%, respectively.

[0042] Example 24 Permeability of prepared solid dispersion of dactinomycin This experiment determined the passive intestinal absorption characteristics of the drug. A 12-well plate (polycarbonate membrane, diameter 12 mm, pore size 0.4 µm, area 1.12 cm 2 ) was used to evaluate the ability of the drug to diffuse from the donor to the receptor chamber. First, 1.5 mL of distilled water was added to the receptor plate. The donor plate was placed above the receptor plate, and 0.5 mL of dactinomycin raw material and drug delivery system sample solutions (equivalent to 1.5 mg / mL of dactinomycin solution) were added to the donor plate, and shaken at 120 rpm in a 37 °C shaker for 210 min. At predetermined time points (30, 60, 90, 120, 150, 180, 210 min), 1 mL of liquid was taken from the receptor chamber and supplemented with the same volume of fresh distilled water in the receptor plate. The sample liquid was filtered through a 0.22 µm water membrane, and the concentration was determined by high performance liquid chromatography using the method described in example 21. The in vivo and in vitro permeation curves were plotted using the concentration points to produce the in vivo and in vitro permeation curves, as shown in Figure 3, the film permeability of the prepared solid dispersion of actinomycin D in Example 1 and Example 2 is enhanced, and the film permeability of the prepared solid dispersion of actinomycin D in Example 1 is better than that in Example 2.

[0043] Example 25 DSC characterization of the prepared solid dispersion of actinomycin D The thermodynamic properties of the raw material, Example 1 and Example 2 were analyzed using SERIES2000 differential scanning calorimeter. The specific method is as follows: 5 mg of the sample to be tested was accurately weighed and placed in an aluminum sample tray, which was tightly sealed. Then the aluminum tray was placed in the detector, and the detection temperature was set to 40-300℃, the temperature rising rate was 10℃ / min, and the detection process was protected by nitrogen. The obtained DSC spectrum is shown in Figure 2. Figure 5 As can be seen from the figure, the melting temperature of actinomycin D is 145℃, and there is an obvious absorption peak at 150℃, which is a sharp peak, indicating that it is the absorption peak of actinomycin. By comparing the DSC curves of Example 1 and Example 2, it can be seen that the characteristic peak of actinomycin in Example 1 almost disappears, and the characteristic peak of actinomycin in Example 2 is significantly reduced, which indicates that after mechanical chemical treatment, the drug has been uniformly dispersed in the carrier, the melting heat of actinomycin D is significantly reduced, and the prepared actinomycin D solid dispersion delivery system has been converted into amorphous state.

[0044] Example 26 XRD characterization of the prepared solid dispersion of actinomycin D The X-ray diffraction patterns of the raw material, Example 1 and Example 2 were recorded using the Empyrean powder diffractometer of the Netherlands PANalytical Company. The specific method is as follows: the sample to be tested was pressed into a flat plane, using a copper radiation source, the voltage was 40KV, the current was 30 mA, the 2θ range was 5-40°, and the scanning angular velocity was 2 deg / min. The obtained XRD spectrum is shown in Figure 3. Figure 4 As can be seen from the figure, actinomycin D has multiple characteristic peaks in the range of 5-40°, indicating its crystalline form. Compared with actinomycin D, the characteristic peaks of Example 2 are significantly reduced, indicating that it has been transformed from crystalline to amorphous. The characteristic peaks of Example 1 almost disappear, which indicates that the degree of amorphization of Example 1 is the highest, which is consistent with the results of DSC. After mechanical ball milling, actinomycin D has been uniformly dispersed in PSP to form a solid dispersion.

[0045] Example 27 Evaluation of the bioavailability of Example The bioavailability of the dactinomycin of Example 1, Example 2 and commercially available dactinomycin (Sigma-Aldrich) in rats was determined according to the following experimental scheme. The rats were subjected to a 12-hour light-dark cycle under controlled temperature and relative humidity, and were allowed to adapt for one week before administration, with free access to food and water. The rats were fasted for 12 hours before administration, with free access to water. The drugs of different formulations and the same dose (20 mg / kg) were prepared into a 1 mg / mL administration concentration with 0.5% sodium carboxymethyl cellulose solution, and were administered to the rats by gavage. Parallel control and self-control were combined to eliminate individual differences of the animals. The rats were subjected to orbital blood collection of 0.7 mL at 0.083, 0.5, 1, 2, 3, 5, 8, 11 and 24 hours after administration, and were carefully mixed in 1.5 mL heparinized centrifuge tubes. Centrifugation was performed at 7000 r / min for 3 min, and the upper plasma was stored in a -20°C refrigerator for 12 hours.

[0046] After the plasma samples were thawed at room temperature in the dark, 200 uL of the plasma sample was accurately transferred into a 1.5 mL centrifuge tube, and 3 mL of 9:1 (v / v) ethyl acetate and hexane was added. After vortexing for 3 min, centrifugation was performed at 7000 rpm / min for 7 min. After centrifugation, the upper organic phase was transferred into another centrifuge tube for vacuum drying. The dried residue was mixed with 100 uL of acetonitrile, vortexed for 1 min, and 10 uL of the mixture was injected into the liquid chromatograph. The method described in Example 21 was used to obtain the content change in the plasma after gavage of dactinomycin of different formulations (as shown in Table 3). Figure 6 The plasma dactinomycin concentration values after gavage of the dactinomycin preparations of the three formulations were input into the DAS 2.0 pharmacokinetic analysis software to obtain the simulated curve of the blood drug concentration and the metabolic kinetic parameters, as shown in Table 2.

[0047] Table 2 Metabolic parameters of the plasma of rats after gavage of dactinomycin of different formulations .

[0048] *p<0.05, **p<0.01 compared with commercially available dactinomycin Note: Cmax: maximum blood drug concentration; Tmax: peak time; T1 / 2(min): half-life; AUC: area under the curve; MRT: mean residence time.

[0049] The experimental conclusion shows that both Example 1 and Example 2 can increase the bioavailability of dactinomycin, which is superior to commercially available dactinomycin.

[0050] Example 28 Evaluation of liver toxicity of dactinomycin The liver toxicity of actinomycin D was evaluated by the method of Example 1 and Example 2, and the test scheme was as follows: the rats were placed in the SPF laboratory animal room (20-25℃, 50-60% relative humidity), and the rats were fed according to the method of Example 11. The rats were divided into four groups, including a normal group, a commercial group, an Example 1 group and an Example 2 group, and each group had five rats. The rats were given physiological saline by gavage, and the amount of actinomycin D in the different formulations was equivalent to 120 mg / kg. After 10 days of gavage, the rats were fasted for 12 h, and blood was taken from the eye sockets into sterile and enzyme-free centrifuge tubes. The supernatant was obtained by centrifugation, and the values of ALT, AST and ALP were detected by a full-automatic biochemical analyzer (Chemray 240) to represent the changes in liver function. The content curves of rat serum ALT, AST and ALP were drawn, as shown in Figure 7 , Figure 8 and Figure 9 From the figure, it can be seen that the values of ALT, AST and ALP of the rats in the Example 1 and commercial groups were significantly reduced, and there was a statistical difference, indicating that Example 1 improved the bioavailability while reducing the damage to the liver. The values of ALT, AST and ALP of the rats in the Example 2 and commercial groups were increased, and there was a statistical difference, indicating that the liver function was damaged, and the use of actinomycin D solid dispersion prepared from polygonatum sibiricum polysaccharide could improve the bioavailability but would aggravate the liver damage, indicating the effectiveness of the formulation of Example 1.

Claims

1. A low-hepatotoxic actinomycin D solid dispersion, characterized in that... The solid dispersion uses a plant-derived functional natural product as a drug carrier and actinomycin D as the drug. Under solvent-free conditions, the plant-derived functional natural product is physically dispersed to form a uniform solid dispersion, which biologically counteracts drug-induced liver damage.

2. The low hepatotoxicity actinomycin D solid dispersion according to claim 1, characterized in that... The plant-derived functional natural product excipients are selected from one of the following: Polyporus umbellatus polysaccharide, Polygonatum sibiricum polysaccharide, mannitol, glycyrrhizic acid, or disodium glycyrrhizate.

3. A method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 1 or 2, characterized in that... The drug actinomycin D, the drug carrier plant-derived functional natural product, the gliding agent, and the grinding beads were placed in a ball mill jar for co-grinding reaction. The grinding was carried out at a speed of 100-400 rpm for 0.5-4 hours to uniformly disperse the drug actinomycin D in the drug carrier plant-derived functional natural product, thereby obtaining a low-hepatotoxicity actinomycin D solid dispersion.

4. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 3, characterized in that... The flow aid is selected from one of micronized silica gel, magnesium stearate, or talc.

5. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 3, characterized in that... The mass ratio of actinomycin D to plant-derived functional natural product materials is 1:1-20.

6. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 3, characterized in that... The mass ratio of actinomycin D to glidin is 20-100:

1.

7. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 3, characterized in that... The diameter of the grinding balls is 5-10mm, and the quantity is 5-10 balls, that is, the ball-to-material mass ratio is 45:1-90:

1.

8. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 3, characterized in that... The ball milling instrument is a planetary ball mill with a rotation speed of 100-400 rpm and a grinding time of 2 hours.

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