Application of fried licorice soup in preparation of drugs for preventing and / or treating cardiotoxicity caused by oxaliplatin

Abstract

The application belongs to the field of medicine and relates to application of a Radix Glycyrrhizae Decoction in preparation of a drug for preventing and / or treating cardiotoxicity caused by oxaliplatin. The application provides a pharmaceutical composition comprising the Radix Glycyrrhizae Decoction, which can be used in preparation of a drug for preventing and / or treating cardiotoxicity caused by oxaliplatin. Experimental results of the application show that the Radix Glycyrrhizae Decoction can improve inflammatory infiltration of myocarditis caused by oxaliplatin, improve myocardial fibrosis caused by oxaliplatin, and improve disorder of myocardial cell arrangement and fibrosis; the Radix Glycyrrhizae Decoction can relieve inflammatory injury of the heart caused by oxaliplatin; and the Radix Glycyrrhizae Decoction can relieve myocardial injury caused by oxaliplatin.

Description

Technical Field

[0001] This invention belongs to the field of medicine and relates to the application of prepared licorice decoction in the preparation of drugs for the prevention and / or treatment of oxaliplatin-induced cardiotoxicity. Background Technology

[0002] Oxaliplatin is a third-generation platinum-based chemotherapy drug with significant advantages in efficacy, pharmacokinetics, antitumor spectrum, and cytotoxicity, and is widely used to treat gastrointestinal malignancies such as colorectal cancer. Currently reported adverse reactions caused by oxaliplatin mainly include peripheral neuropathy, allergic reactions, and gastrointestinal reactions. Furthermore, recent clinical cases have reported ST-segment elevation and QT interval prolongation with torsades de pointes in patients treated with oxaliplatin, indicating that its cardiotoxicity should not be ignored. Therefore, there is an urgent need to develop a drug that can prevent and / or treat oxaliplatin-induced cardiotoxicity.

[0003] Traditional Chinese medicine categorizes the cardiotoxicity caused by chemotherapy drugs as "drug toxicity." The *Zhu Bing Yuan Hou Lun* (Treatise on the Causes and Symptoms of Various Diseases) states, "All drugs that are said to be toxic or highly toxic can cause disturbances, harm people, and even kill them." The Zhigancao Tang (Prepared Licorice Decoction) originates from Zhang Zhongjing's *Shang Han Lun* (Treatise on Cold Damage) of the Eastern Han Dynasty. In this formula, prepared licorice root has the effects of sweet and warming, invigorating qi, unblocking meridians, and promoting blood circulation; ginseng and jujube have the effects of tonifying qi and benefiting the stomach, nourishing the source of blood vessels; donkey-hide gelatin, rehmannia root, ophiopogon root, and hemp seed are used to nourish heart blood and replenish heart yin; cinnamon twig and ginger are added for their pungent and warming properties to promote heart yang. The combined effects of these herbs are to nourish yin and qi, promote yang, and restore blood vessels. There are no reports of its protective effect against oxaliplatin-induced cardiotoxicity. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to address the shortcomings of the prior art by providing the application of prepared licorice decoction in the preparation of drugs for the prevention and / or treatment of cardiotoxicity caused by oxaliplatin.

[0005] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows:

[0006] This invention discloses the application of prepared licorice decoction in the preparation of drugs for the prevention and / or treatment of oxaliplatin-induced cardiotoxicity;

[0007] The prepared licorice decoction is made from the following components in parts by weight:

[0008] Prepared licorice root 10-14 parts,

[0009] 7-11 parts ginger

[0010] 5-7 parts ginseng

[0011] 45-50 parts of raw Rehmannia glutinosa

[0012] 8-10 parts of cinnamon twigs

[0013] 5-7 parts of donkey-hide gelatin

[0014] 8-12 portions of Ophiopogon japonicus.

[0015] 8-12 parts hemp seeds

[0016] 22-27 portions of jujubes.

[0017] In some embodiments, preferably, the prepared licorice decoction comprises the following components in parts by weight:

[0018] 12 parts of roasted licorice root,

[0019] 9 portions of ginger

[0020] 6 portions of ginseng

[0021] 48 portions of raw Rehmannia glutinosa

[0022] Nine portions of cinnamon twigs.

[0023] 6 parts of donkey-hide gelatin,

[0024] 10 portions of Ophiopogon japonicus

[0025] 10 portions of hemp seeds

[0026] 25 portions of jujubes.

[0027] In some embodiments, more preferably, the prepared licorice decoction is made from the following components in parts by weight: 12g prepared licorice root, 9g ginger, 6g ginseng, 48g rehmannia root, 9g cinnamon twig, 6g donkey-hide gelatin, 10g ophiopogon root, 10g hemp seed, and 25g jujube.

[0028] In some embodiments, the prepared licorice decoction is prepared by conventionally decocting a mixture of prepared licorice, ginger, ginseng, rehmannia root, cinnamon twig, ophiopogon root, hemp seed, jujube, wine, and water in the indicated weight proportions, and then adding the indicated weight proportions of donkey-hide gelatin to dissolve it.

[0029] The wine is preferably glutinous rice wine; the water is preferably double-distilled water; the volume ratio of wine to water is 6-8:7-9, preferably 7:8; the total weight of the prepared licorice root, ginger, ginseng, rehmannia root, cinnamon twig, ophiopogon root, hemp seed, jujube and donkey-hide gelatin to water is 125-145:1500-1700.

[0030] In a further preferred embodiment, 12g of roasted licorice root, 9g of ginger, 6g of ginseng, 48g of rehmannia root, 9g of cinnamon twig, 10g of ophiopogon root, 10g of hemp seed, and 25g of jujube are mixed and soaked in 1400mL of glutinous rice wine and 1600mL of double-distilled water. The mixture is first heated to a boil over high heat, then simmered over low heat until the decoction reaches about 1000mL. Then, 6g of donkey-hide gelatin is added and melted, and the mixture is concentrated to obtain roasted licorice root decoction.

[0031] The concentration of the raw medicinal material in the concentrated licorice decoction is not particularly limited, but it is preferably concentrated to a concentration of 2.0-4.0 g / mL, and more preferably 3.1 g / mL.

[0032] In some embodiments, the prevention and / or treatment of oxaliplatin-induced cardiotoxicity is to improve oxaliplatin-induced myocardial inflammatory infiltration.

[0033] In some embodiments, the prevention and / or treatment of oxaliplatin-induced cardiotoxicity is to improve oxaliplatin-induced myocardial fibrosis.

[0034] In some embodiments, the prevention and / or treatment of oxaliplatin-induced cardiotoxicity is to improve oxaliplatin-induced cardiomyocyte disorder.

[0035] In some embodiments, the prevention and / or treatment of oxaliplatin-induced cardiotoxicity is to improve oxaliplatin-induced cardiac inflammatory damage.

[0036] In some embodiments, the prevention and / or treatment of oxaliplatin-induced cardiotoxicity is to improve oxaliplatin-induced myocardial damage.

[0037] Beneficial effects:

[0038] (1) The present invention provides a pharmaceutical composition comprising roasted licorice root decoction, which can be used to prepare a drug for preventing and / or treating cardiotoxicity caused by oxaliplatin.

[0039] (2) The experimental results of this invention show that the prepared licorice decoction can improve the myocardial inflammatory infiltration caused by oxaliplatin in rats, improve the myocardial fibrosis caused by oxaliplatin in rats, and improve the disordered arrangement and fibrosis of myocardial cells; the prepared licorice decoction can alleviate the cardiac inflammatory damage caused by oxaliplatin; the prepared licorice decoction can alleviate the myocardial damage caused by oxaliplatin. Attached Figure Description

[0040] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, and the advantages of the present invention in the above and / or other aspects will become clearer.

[0041] Figure 1 Representative HE staining images of heart tissue from each group of rats.

[0042] Figure 2 Representative images of Masson staining and Sirius red staining of rat heart tissue from each group.

[0043] Figure 3 The expression levels of serum inflammatory factors in rats of each group; among them, ** P < 0.01, **** P < 0.0001.

[0044] Figure 4 The expression levels of serum myocardial injury markers in each group of rats; among them, * P < 0.05 *** P < 0.001. Detailed Implementation

[0045] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0046] Example 1

[0047] 1. Experimental materials

[0048] (1) Animals: Female SPF-grade healthy Wistar rats, weighing 200±20g.

[0049] (2) Main reagents and equipment: Oxaliplatin (OXA), ZGCT.

[0050] 2. Experimental Procedure

[0051] 2.1 Reagent Preparation

[0052] Preparation of ZGCT (Prepared Licorice Root Decoction): Prepared licorice root 12g, fresh ginger 9g, ginseng 6g, rehmannia root 48g, cinnamon twig 9g, donkey-hide gelatin 6g, ophiopogon root 10g, hemp seed 10g, jujube 25g, totaling 135g. The herbs were purchased from the Traditional Chinese Medicine Pharmacy of Jiangsu Provincial Hospital of Integrated Traditional and Western Medicine. According to the method recorded in *Shang Han Lun* (Treatise on Cold Damage): "Use seven liters of clear wine and eight liters of water to first boil the eight herbs, then reduce to three liters. Remove the dregs, add the donkey-hide gelatin and dissolve completely. Take one liter warm, three times a day." Mix the eight herbs except for the donkey-hide gelatin. For each dose, add 1400mL of glutinous rice wine and 1600mL of double-distilled water to soak. First, heat over high heat to a boil, then reduce to a simmer until the decoction reaches about 1000mL. Add the donkey-hide gelatin and dissolve. Then, concentrate each dose using a rotary evaporator to a raw herb content of 3.1g / mL. After cooling, store in a 4℃ refrigerator for later use.

[0053] Preparation of oxaliplatin (OXA) solution: Dissolve 50 mg of oxaliplatin in 25 mL of 5% glucose solution to a final concentration of 2 mg / mL, and store at 4°C for later use. Dilute with 5% glucose solution to a working solution of 0.4 mg / mL immediately before use.

[0054] 2.2 Animal grouping and administration

[0055] After one week of acclimatization, 24 Wistar rats were randomly divided into 3 groups of 8 rats each: solvent control group (Vehicle group), model group (OXA group), and licorice decoction group (ZGCT group).

[0056] Except for the Vehicle group, the other two groups were used to establish a rat model of cardiac injury by intraperitoneal injection of 0.4 mg / mL OXA working solution (i.e., the OXA dosage was 4 mg / kg) at a dose of 10 mL / kg on days 1, 2, 8, 9, 15, 16, 22, and 23. The Vehicle group was injected intraperitoneally with an equal volume of 5% glucose solution. Starting from day 1, the ZGCT group was administered 12 g / kg of Zhigancao decoction (clinically equivalent dose) by gavage, while the Vehicle and OXA groups were administered an equal volume of double-distilled water by gavage, once daily for 32 consecutive days. When Zhigancao decoction was administered in combination with OXA, each group was administered 0.5 hours before the administration of OXA (i.e., 0.4 mg / mL OXA working solution) / 5% glucose solution by gavage.

[0057] 3. Specimen Collection and Processing

[0058] On day 33, rats were anesthetized, and blood samples were collected via the abdominal aorta. The rats were then euthanized by cervical dislocation, and their hearts were dissected and fixed in 4% paraformaldehyde fixative. After the blood was allowed to stand for 2 hours, it was centrifuged at 3500 rpm for 15 minutes, and the supernatant was collected, aliquoted, and stored at -80℃ for later use.

[0059] The expression levels of interleukin-6 (IL-6), creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), and N-terminal pro-Brain natriuretic peptide (NT-proBNP) in serum were detected by ELISA. Histopathological changes in rat myocardial tissue were observed by hematoxylin-eosin (HE) staining, Masson staining, and Sirius red staining.

[0060] 4. Statistical processing

[0061] Statistical analysis and graphing were performed using GraphPad Prism 9.0 software. Data are expressed as mean ± standard error (SEM). One-way ANOVA was used for comparisons among multiple groups. A p-value < 0.05 was considered statistically significant.

[0062] 5. Results

[0063] 5.1. The use of prepared licorice root decoction alleviated oxaliplatin-induced myocardial pathological damage in rats.

[0064] HE staining was performed on sections of rat heart tissue to observe the morphology and structure of cardiomyocytes. The results are as follows: Figure 1As shown, in the Vehicle group, the cardiomyocyte nuclei of rats were normal in size, regular in shape, and arranged in an orderly manner. Compared with the Vehicle group, the cardiomyocyte nuclei of rats in the OXA group were aggregated, and the cardiomyocytes were irregular in shape and disordered in arrangement, showing inflammatory infiltration. Compared with the OXA group, the cardiomyocytes of rats in the ZGCT group were relatively regular in shape and clearer in structure, suggesting that the licorice decoction can improve the inflammatory infiltration of the rat myocardium caused by OXA.

[0065] 5.2. The use of prepared licorice root decoction to alleviate oxaliplatin-induced myocardial fibrosis in rats.

[0066] Masson staining and Sirius red staining were performed on rat heart tissue sections to observe myocardial fibrosis. The results are as follows: Figure 2 As shown.

[0067] In Masson staining, blue represents fibrotic connective tissue. The image shows that the cardiomyocytes in the Vehicle group are in normal condition, with orderly cell arrangement and only a small amount of collagen evenly distributed between the myocardial fibers. In contrast, the OXA group showed a significant increase in myocardial collagen fiber deposition, with abundant collagen distributed between myocardial fibers and interconnected in a network, exhibiting disordered and uneven distribution, suggesting that OXA induced myocardial fibrosis and remodeling. Compared to the OXA group, the ZGCT group improved oxaliplatin-induced myocardial fibrosis in rats.

[0068] In Sirius red staining, normal cardiomyocytes appear orange, while fibrous tissue appears red. As shown in the figure, the cardiomyocytes in the Vehicle group were neatly arranged, while the red fibrous tissue in the OXA group was significantly increased; compared to the OXA group, the ZGCT group improved cardiomyocyte disorder and fibrosis.

[0069] 5.3. The decoction of prepared licorice root reduced serum inflammatory factor levels in rats.

[0070] The expression level of IL-6 in the serum of rats in each group was detected by ELISA, and the results are as follows: Figure 3 As shown. Compared to the Vehicle group, the serum IL-6 expression level in the OXA group rats was significantly increased ( **** P < 0.0001, suggesting that oxaliplatin can induce inflammatory damage; while compared with the OXA group, IL-6 expression was significantly decreased in the ZGCT group (P < 0.0001). ** P < 0.01, suggesting that Zhigancao Decoction can alleviate inflammatory damage caused by oxaliplatin.

[0071] 5.4. Prepared licorice root decoction reduced the levels of serum myocardial injury markers in rats.

[0072] ELISA was used to further detect the expression levels of myocardial injury markers CK-MB, LDH, and NT-proBNP in the serum of rats in each group. The results are as follows: Figure 4 As shown in the figure, compared to the Vehicle group, the serum expression levels of CK-MB, LDH, and NT-proBNP in the OXA group were significantly increased, suggesting that oxaliplatin has cardiotoxicity; while compared to the OXA group, the expression levels of CK-MB, LDH, and NT-proBNP in the ZGCT group were significantly decreased, suggesting that the herbal decoction can alleviate oxaliplatin-induced myocardial damage. * P < 0.05 *** P < 0.001 indicates a significant difference.

[0073] This invention provides a concept and method for the application of prepared licorice root decoction in the preparation of drugs for the prevention and / or treatment of oxaliplatin-induced cardiotoxicity. Many methods and approaches exist for implementing this technical solution; the above description is merely a preferred embodiment of the invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications should also be considered within the scope of protection of this invention. All components not explicitly stated in this embodiment can be implemented using existing technologies.

Claims

1. The application of Zhigancao Decoction in the preparation of drugs for the prevention and / or treatment of oxaliplatin-induced cardiotoxicity; The prepared licorice decoction is made from the following components in parts by weight: Prepared licorice root 10-14 parts, 7-11 parts ginger 5-7 parts ginseng 45-50 parts of raw Rehmannia glutinosa 8-10 parts of cinnamon twigs 5-7 parts donkey-hide gelatin, 8-12 portions of Ophiopogon japonicus. 8-12 parts hemp seeds 22-27 servings of jujubes; The prevention and / or treatment of oxaliplatin-induced cardiotoxicity is to improve oxaliplatin-induced myocardial inflammatory infiltration, myocardial fibrosis, cardiomyocyte disorder, cardiac inflammatory damage, or myocardial injury.

2. The application according to claim 1, characterized in that, The prepared licorice decoction is made from the following components in parts by weight: 12 parts prepared licorice root, 9 parts fresh ginger, 6 parts ginseng, 48 parts raw rehmannia root, 9 parts cinnamon twig, 6 parts donkey-hide gelatin, 10 parts ophiopogon root, 10 parts hemp seed, and 25 parts jujube.

3. The application according to claim 1, characterized in that, The prepared licorice decoction is made by conventionally decocting a mixture of prepared licorice, ginger, ginseng, rehmannia root, cinnamon twig, ophiopogon root, hemp seed, jujube, wine, and water in the specified weight proportions, and then adding the specified weight proportions of donkey-hide gelatin to dissolve it.

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