A low hepatotoxicity dactinomycin solid dispersion and a preparation method thereof

Abstract

The present application relates to a low hepatotoxicity actinomycin D solid dispersion and a preparation method thereof. The low hepatotoxicity actinomycin D solid dispersion product is obtained by co-grinding reaction of actinomycin D, plant source functional natural excipient, flow aid and ball milling beads in a ball milling tank. The solid dispersion is prepared by mechanical chemical grinding, without using organic solvent, avoiding drug degradation in high temperature melting process, easily making the drug and excipient amorphous, improving the drug solubility and bioavailability, having the advantages of high preparation efficiency, low production cost, simple operation, green environmental protection and the like; and the plant source polymer is used as a carrier, the solubilization and synergistic effect are enhanced, the hepatotoxicity of actinomycin D is reduced, which is crucial for safe use of antibiotic drugs, and has positive significance for exploring natural, non-toxic, efficient and multifunctional drug loading system excipient.

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, that is, 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)

[0012] Further, the application also limits the ball milling instrument to be a planetary ball mill, the rotating speed of which is 100-400 rpm, and the grinding time is 2 h.

[0013] By using the above-mentioned technology, the application has the following beneficial effects compared with the prior art:

[0014] 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 achieved by “drug-assisted drug”, that is, 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 it physically, 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 achieved, the bioavailability of the antitumor drug is improved, and the liver toxicity caused by the drug is reduced by the carrier itself.

[0015] 2) The preparation process of the present application discards a large amount of organic solvents, and solves the problems of solvent residue, environmental pollution, complex process and the risk of drug degradation. The mechanical 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 meets the development trend of modern pharmaceutical industry;

[0016] 3) The solid dispersion obtained by the present application significantly improves the solubility and bioavailability of actinomycin D, effectively reduces its fatal hepatotoxicity by using the liver-protecting effect of the active carrier, widens the therapeutic window, and significantly improves the safety. The improvement of bioavailability means that the required dose for the same efficacy may be reduced, further reducing the risk of toxic side effects, achieving the clinical goal of "attenuation and efficiency", thus increasing the efficacy. The operation is simple, avoids the use of organic solvents, prevents solvent residue and problems such as raw material loss and thermal decomposition caused by solvent thermal removal process, has the advantages of high preparation efficiency, safe and reliable production, low production cost, less pollution, etc., is easier to industrialize, opens up a new way for the application of active ingredients (polysaccharides) in modern high-value drug preparations, and promotes the in-depth development of the combination of traditional Chinese medicine and Western medicine. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 is the actinomycin D dissolution curve in a medium with pH = 1.2;

[0018] Figure 2 is the actinomycin D dissolution curve in a medium with pH = 6.8;

[0019] Figure 3 is the in vitro membrane permeation curve of actinomycin D;

[0020] Figure 4 is the X-ray diffraction spectrum of actinomycin D, example 1 and example 2;

[0021] Figure 5 is the differential scanning calorimetry spectrum of actinomycin D, example 1 and example 2;

[0022] Figure 6 is the drug content change curve in the plasma of rats after gavage with different actinomycin D;

[0023] Figure 7 is the serum ALT content graph of rats after gavage with different actinomycin D;

[0024] Figure 8 is the serum AST content graph of rats after gavage with different actinomycin D;

[0025] Figure 9 is the serum ALP content graph of rats after gavage with different actinomycin D. DETAILED DESCRIPTION

[0026] The application will be further described in conjunction with specific examples, but the scope of protection of the application is not limited to this.

[0027] Example 1 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, krestin 700 mg, micro-silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0028] Example 2 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, rhizoma polygonati polysaccharide 700 mg, micro-silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0029] Example 3 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, mannitol 700 mg, micro-silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0030] Example 4 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, glycyrrhizic acid 700 mg, micro-silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0031] Example 5 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, disodium glycyrrhizinate 700 mg, micro-silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0032] Example 6 Into a 50 mL polytetrafluoroethylene ball mill jar, add actinomycin D 100 mg, krestin 700 mg, magnesium stearate 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an actinomycin D solid dispersion.

[0033] Example 7 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, talc 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0034] Example 8 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 300 mg, microfine silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0035] Example 9 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 1000 mg, microfine silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0036] Example 10 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 1500 mg, microfine silica gel 2 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0037] Example 11 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, microfine silica gel 1 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0038] Example 12 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, microfine silica gel 5 mg, and 8 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0039] Example 13 Into a 50 mL polytetrafluoroethylene ball mill jar, add Actinomycin D 100 mg, Krestin 700 mg, microfine silica gel 2 mg, and 5 stainless steel ball milling beads with a diameter of 7.5 mm in sequence, and mill for 2 h at 200 rpm. After ball milling, the composite powder is taken out to obtain an Actinomycin D solid dispersion.

[0040] Example 14 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 10 pieces, 7.5 mm in diameter, sequentially, and ground at 200 rpm for 2 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0041] Example 15 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 8 pieces, 7.5 mm in diameter, sequentially, and ground at 200 rpm for 0.5 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0042] Example 16 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 8 pieces, 7.5 mm in diameter, sequentially, and ground at 200 rpm for 1 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0043] Example 17 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 8 pieces, 7.5 mm in diameter, sequentially, and ground at 200 rpm for 4 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0044] Example 18 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 10 pieces, 7.5 mm in diameter, sequentially, and ground at 100 rpm for 2 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0045] Example 19 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 8 pieces, 7.5 mm in diameter, sequentially, and ground at 300 rpm for 2 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0046] Example 20 To a 50 mL polytetrafluoroethylene grinding jar was added Actinomycin D 100 mg, Krestin 700 mg, finely powdered silica gel 2 mg, and stainless steel grinding beads 8 pieces, 7.5 mm in diameter, sequentially, and ground at 400 rpm for 2 h. After the grinding, the complex powder was taken out to obtain an Actinomycin D solid dispersion.

[0047] Example 21 Actinomycin D standard curve preparation:

[0048] Precisely take 20 mg of actinomycin D standard (source leaf, 98%) and place it in a 50 mL volumetric flask. Add mobile phase to dissolve, dilute to the mark, sonicate to dissolve, and shake well. 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 into 9 10 mL volumetric flasks, dilute with mobile phase to the mark, shake well, and dilute to 4, 20, 40, 100, 120, 140, 160, 200, 240 ug / mL, respectively. Filter through a 0.22 μm filter and 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.

[0049] Obtain the peak areas corresponding to different concentrations of actinomycin D, and perform linear regression of the peak areas versus the concentrations. The regression equation is y = 19891x - 12793 (R 2 = 0.9999) with an applicable concentration range of (4 μg / mL - 250 μg / mL).

[0050] Dilute the solution to be determined to within the peak area range of the standard curve, and record the dilution factor as V. Determine the peak area PA of the solution by HPLC, and calculate the concentration c (ug / mL) of actinomycin D in the solution to be determined according to the following formula.

[0051] Formula 1: c (ug x mL -1 ) = V x (PA + 2793) ÷ 19891.

[0052] Example 22 Determination of the solubility and preparation yield of actinomycin D prepared by the examples

[0053] The method is as follows: an excess of the prepared solid dispersion of actinomycin D is weighed, placed in a conical flask, and added to 20 ml of distilled water. The solution is shaken to reach saturation concentration. The shaking parameters are a temperature of 37°C, a rotation speed of 200 rpm, and a time of 24 h. The saturated solution is filtered using a 0.22 μm filter membrane, and the peak area PA is determined using HPLC. The concentration of actinomycin D in the saturated solution in water is calculated 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, which is the actual product amount / theoretical product amount*100%. The equilibrium solubility and yield of each example are shown in Table 1. The results show that the solubilization effect of the actinomycin D solid dispersions in Example 1 and Example 2 is better, and the powder flowability is good and the preparation yield is high. Examples 1 and 2 are selected for further physicochemical property and pharmacodynamic characterization.

[0054] Table 1: Solubility and preparation yield parameters of each formulation

[0055] .

[0056] Dissolution of the prepared actinomycin D solid dispersion in Example 23

[0057] The in vitro dissolution rate of the sample is tested according to the second method (paddle method) of the Chinese Pharmacopoeia 2020 edition. The theoretical drug content of 50.0 mg of actinomycin D raw material and the drug delivery system of Examples 1 and 2 are weighed, and potassium dihydrogen phosphate and sodium hydroxide are used to prepare pH=1.2 and pH=6.8 phosphate buffer solutions as dissolution liquids. The temperature of the dissolution instrument is set to 37.0±0.5°C, and the paddle rotation speed is 75 rpm. The sample solution is taken at 5, 10, 15, 20, 30, 40, 60, 90, 120, 150, and 180 min, and 5 mL of fresh dissolution medium is immediately added. The sample solution is filtered through a 0.22 µm water membrane, and the concentration is determined by high performance liquid chromatography using the method described in Example 21. The dissolution curve is plotted using the concentration points, and the dissolution curves are shown in Figure 1 and Figure 2 . As shown in Figure 1 and Figure 2 , whether in pH=1.2 or pH=6.8 dissolution medium, the release rate and dissolution effect of actinomycin D are significantly improved. Among them, the final dissolution of Example 1 in pH=1.2 and pH=6.8 dissolution medium is increased by about 38% and 60%, respectively.

[0058] Permeability of the prepared actinomycin D solid dispersion in Example 24

[0059] The passive intestinal absorption characteristics of the drug were determined using a 12-well plate (polycarbonate membrane, diameter 12 mm, pore size 0.4 μm, area 1.12 cm 2 The ability of the drug to diffuse from the donor to the receiver chamber was evaluated. First, 1.5 mL of distilled water was added to the receiver plate. The donor plate was placed above the receiver plate, 0.5 mL of the sample solution of actinomycin D raw material and the drug delivery system examples 1 and 2 (equivalent to 1.5 mg / mL of actinomycin D solution) was added to the donor plate, and shaken at 120 rpm in a 37 °C shaker for 210 min. At the predetermined time points (30, 60, 90, 120, 150, 180, 210 min), 1 mL of liquid was taken from the receiver chamber, and the same volume of fresh distilled water was added to the receiver plate. The sample solution 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, and the peak area was converted to concentration. The in vitro permeability curve was plotted using the concentration points, and the in vitro permeability curve is shown in Figure 3 The membrane permeability of examples 1 and 2 was better than that of the commercially available actinomycin D.

[0060] Example 25 DSC characterization of the prepared actinomycin D solid dispersion

[0061] The thermodynamic properties of the raw material, example 1 and example 2 were analyzed using a SERIES 2000 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 °C, the heating rate was 10 °C / min, and the nitrogen was protected during the detection process. The obtained DSC spectrum is shown in Figure 5 As can be seen from the figure, the melting temperature of actinomycin D is 145 °C, and there is an obvious absorption peak at 150 °C, which is a sharp peak, indicating that it is the absorption peak of actinomycin. By comparing the DSC curves of examples 1 and 2, it can be seen that the characteristic peak of example 1 actinomycin almost disappears, and the characteristic peak of 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 to amorphous state.

[0062] Example 26 XRD characterization of the prepared actinomycin D solid dispersion

[0063] The X-ray diffraction patterns of the raw material, Example 1, and Example 2 were recorded using an Empyrean powder diffractometer from 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, a voltage of 40KV, a current of 30mA, a 2θ range of 5-40°, and an angular velocity of scanning of 2 deg / min. The obtained XRD spectrum is shown in Figure 1. Figure 4 As can be seen from the figure, there are multiple characteristic peaks of actinomycin D in the range of 5-40°, indicating that it is in a crystalline state. Compared with actinomycin D, the characteristic peaks of Example 2 are significantly reduced, indicating that it has been transformed from a crystalline state to an amorphous state. The characteristic peaks of Example 1 have almost disappeared, indicating that Example 1 has the highest degree of amorphization, 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.

[0064] Example 27 Evaluation of the bioavailability of Example

[0065] The bioavailability of Example 1, Example 2, and commercially available actinomycin D (Sigma-Aldrich Company) in rats was determined, and the experimental scheme was as follows: the rats were subjected to a 12-hour light-dark cycle under controlled temperature and relative humidity, and were adapted for one week before administration, and were allowed to eat and drink freely. The rats were fasted for 12h before administration, and were allowed to drink freely. Different formulations of the same dose of drug (20mg / kg) were prepared into a 1mg / mL administration concentration with 0.5% sodium carboxymethyl cellulose solution, and were administered to rats by gavage. Parallel control and self-control were combined to eliminate individual differences among animals. At 0.083, 0.5, 1, 2, 3, 5, 8, 11, and 24h after administration, 0.7mL of blood was taken from the orbital cavity of the rats, and was carefully mixed in a 1.5ml heparinized centrifuge tube. After centrifugation at 7000r / min for 3min, the upper layer of plasma was taken and stored in a-20℃ refrigerator for 12h.

[0066] After the plasma sample was thawed at room temperature in the dark, 200uL of the plasma sample was precisely transferred into a 1.5mL centrifuge tube, and 3mL of 9:1(v / v) ethyl acetate and hexane was added. After vortexing for 3min, the mixture was centrifuged at 7000rpm / min for 7min. After centrifugation, the upper organic phase was transferred to another centrifuge tube for vacuum drying. The dried residue was mixed with 100uL of acetonitrile, vortexed for 1min, and 10uL of the mixture was injected into the liquid chromatograph. Using the method described in Example 21, the content changes in the plasma after gavage of actinomycin D of different formulations were obtained (as shown in Table 1). Figure 6 The plasma actinomycin D concentration values after gavage of the three dosage forms of actinomycin D 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.

[0067] Table 2 Metabolic parameters of different prescriptions of dactinomycin in rats

[0068] .

[0069] *p<0.05, **p<0.01 compared with commercially available dactinomycin

[0070] Note: Cmax: maximum plasma concentration; Tmax: peak time; T1 / 2(min): half-life; AUC: area under the curve; MRT, mean residence time.

[0071] The experimental results show that both Example 1 and Example 2 can increase the bioavailability of dactinomycin, which is better than commercially available dactinomycin.

[0072] Example 28 Evaluation of dactinomycin hepatotoxicity

[0073] The hepatotoxicity of Example 1 and Example 2 was evaluated by reducing dactinomycin. The test scheme is as follows: the rats were placed in the SPF experimental animal room (20~25℃, 50~60% relative humidity), and the rats were fed according to the method of Example 11. Different prescriptions of dactinomycin corresponding to 120mg / kg dactinomycin were given by gavage, divided into four groups, including normal group, commercial group, Example 1 group and Example 2 group, five rats in each group, gavage for 10 days, after fasting for 12h, the blood was taken from the eye socket into a sterile and enzyme-free centrifuge tube, centrifuged, and the supernatant was taken to detect the values of ALT, AST and ALP by automatic biochemical analyzer (Chemray240) to represent the changes of liver function, and the content curve of rat serum ALT, AST and ALP was 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 rats in Example 1 and commercial group are significantly reduced, and there is a statistical difference, which shows that Example 1 can improve the bioavailability while reducing the damage to the liver; while the values of ALT, AST and ALP of rats in Example 2 and commercial group are increased, and there is a statistical difference, which shows that the liver function is damaged, and the use of dactinomycin solid dispersion prepared by Polygonatum sibiricum can improve the bioavailability, but will aggravate the liver damage, which shows that the formula of Example 1 is effective.

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 through mechanical grinding. Biologically, it counteracts drug-induced liver damage. The plant-derived functional natural product is Polyporus umbellatus polysaccharide.

2. A method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 1, 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 at a speed of 100-400 rpm for 0.5-4 hours, so that the drug actinomycin D was uniformly dispersed in the drug carrier plant-derived functional natural product, and a low hepatotoxic actinomycin D solid dispersion was obtained.

3. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 2, characterized in that... The flow aid is selected from one of micronized silica gel, magnesium stearate, or talc.

4. The method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 2, characterized in that... The mass ratio of actinomycin D to plant-derived functional natural product materials is 1:1-20.

5. A method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 2, characterized in that... The mass ratio of actinomycin D to the descent agent is 20-100:

1.

6. A method for preparing a low-hepatotoxicity actinomycin D solid dispersion according to claim 2, 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.

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