New use of Xintong preparation in preparation of medicine for treating anemia

Abstract

The present application relates to the technical field of traditional Chinese medicine preparation, in particular to a new application of Xintong preparation in preparing anemic drugs. The present application discloses the clinical application of Xintong preparation composed of 13 kinds of traditional Chinese medicines, such as Huangqi, Dangshen, Maidong, Heshouwu, Yinyanghe, Gegen, Dangui, Danshen, Zaokesheng, Haizao, Kunkunbu, Muli and Zhishi, in treating anemia caused by qi stagnation and blood stasis and phlegm-dampness blockage. Experiments show that the Xintong preparation can significantly improve the hemoglobin level of anemic model rats, and has better effects than the positive control in blood sugar control and kidney tissue repair. The present application expands the clinical application range of Xintong preparation, provides a multi-target treatment scheme for refractory anemia combined with microcirculation disorder and metabolic disorder, and has significant clinical value and innovation.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine preparation, in particular to a new use of Xintong preparation in the preparation of an anemia treating medicine. BACKGROUND

[0002] Anemia is one of the common blood system diseases in the world, which is characterized by a decrease in the number of red blood cells or a decrease in the concentration of hemoglobin, leading to a decrease in tissue oxygen carrying capacity, causing symptoms such as fatigue, palpitation, and multiple organ dysfunction.

[0003] According to the etiology, anemia is divided into iron deficiency anemia, megaloblastic anemia, chronic anemia, renal anemia and other types by western medicine. Among them, chronic anemia is often secondary to chronic inflammatory diseases such as rheumatoid arthritis, metabolic diseases such as diabetes, cardiovascular diseases such as coronary heart disease, or chronic kidney disease, and its core pathological mechanism involves the interaction of microcirculation disorder, vascular endothelial damage, metabolic disorder and nutritional absorption disorder. The treatment of western medicine mainly supplements iron and erythropoietin, but the effect is limited, especially for chronic anemia with microcirculation disorder.

[0004] According to the theory of traditional Chinese medicine, anemia is classified into "blood deficiency" and "deficiency". The classic prescription such as angelica sinensis blood tonic soup and spleen soup takes blood production as the core, and has definite effect on simple anemia with deficiency of both qi and blood. However, about 40% of anemia patients in clinical practice have "deficiency of root and excess of branch" pathogenesis, that is, deficiency of both qi and blood combined with stagnation of qi and blood stasis, phlegm and dampness obstruction. This type of anemia often shows symptoms such as dark complexion, purple moss on the tongue, and pulse contraction, which is highly related to microcirculation disorder, coronary heart disease combined with anemia, and other chronic inflammatory states such as rheumatoid arthritis. The traditional Chinese medicine treatment mainly focuses on tonifying qi and blood or invigorating the spleen to remove dampness, but there is a lack of compound preparation with the functions of tonifying qi and activating blood and removing phlegm and dredging collaterals, especially the targeted regulation drugs for microcirculation and vascular endothelial function, which are prone to "tonifying stagnation" and lack of correlation with western medicine pathological indicators such as vascular endothelial factor, inflammatory markers and metabolic parameters, making it difficult to meet the evidence-based requirements of modern medical research and development.

[0005] Therefore, it is urgent to develop effective drugs for anemia caused by such specific causes. SUMMARY

[0006] In view of the treatment defect of limited clinical effect in the prior art for anemia combined with stagnation of qi and blood stasis, phlegm and dampness obstruction, the present application provides the use of Xintong preparation in the preparation of an anemia treating medicine, which is especially suitable for refractory anemia secondary to chronic diseases or metabolic disorders.

[0007] Xintong preparation, approval number: GMP, is a protected Chinese medicine variety produced by Lunan Houpu Pharmaceutical Co., Ltd., which is composed of 13 kinds of Chinese medicines such as Huangqi, Dangshen, Maidong, Heshouwu, Yinyanghe, Gegen, Dangui, Danshen, Zaokesheng, Haizao, Kelp, Muli and Zhishi, has the effects of benefiting qi and nourishing blood, removing blood stasis and dredging collaterals, and is used for treating chest stuffiness, heartache, palpitation, chest distress, shortness of breath, restlessness and fatigue, and pulse is sinking, fine, stringy, slippery or bound.

[0008] The inventor found that Xintong preparation has a significant effect on treating a specific type of anemia in clinical practice.

[0009] Specifically, the technical scheme of the present application is as follows:

[0010] The use of Xintong preparation in preparing an anemia treatment medicine.

[0011] Specifically, the anemia is caused by blood stasis and phlegm dampness.

[0012] The anemia is caused by at least one factor of microcirculation disorder, vascular endothelial dysfunction or metabolic disorder, and the anemia can be combined with chronic inflammation, tissue ischemia or nutritional absorption disorder, and the present application also covers the case that the anemia is secondary to rheumatoid arthritis, chronic kidney disease, coronary heart disease, diabetes, hyperlipidemia or non-alcoholic fatty liver disease.

[0013] Xintong preparation can be used as the only medicine in preparing an anemia treatment medicine, or can be used in combination with qi and blood tonics in preparing an anemia treatment medicine, thereby widening its application range and treatment effect.

[0014] Specifically, the use of Xintong preparation in combination with Danggui Buxue Decoction, Guipi Pill and Colla Corii Asini preparation in preparing an anemia treatment medicine is suitable for the case of severe blood stasis and deficiency of both qi and blood.

[0015] The Xintong preparation of the present application is prepared from 13 kinds of raw medicinal materials, i.e., Huangqi, Dangshen, Maidong, Heshouwu, Yinyanghe, Gegen, Dangui, Danshen, Zaokesheng, Haizao, Kelp, Muli and Zhishi, in a certain ratio.

[0016] One preferred ratio is as follows:

[0017]

[0018] The dosage form of the Xintong preparation of the present application can be Xintong granules or Xintong oral liquid, which is convenient for patients to take and absorb, and is beneficial to improving the compliance and treatment effect of the medicine.

[0019] Compared with the prior art, the present application has the following advantages:

[0020] 1, The heart pass preparation is first found to have a significant curative effect on refractory anemia of "qi stagnation and blood stasis, phlegm dampness obstruction", and the anemia is often secondary to chronic diseases such as rheumatoid arthritis, chronic kidney disease, diabetes and other chronic diseases with microcirculation disorder and metabolic disorder, which provides a new treatment choice for clinic.

[0021] 2, The heart pass preparation is a clinical application of traditional Chinese medicine compound (Z10920014), which has convenient dosage form, higher safety and significant clinical effect, compared with western medicine EPO for injection or long-term iron agent, the inconvenience of injection and the risk of iron overload are avoided. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 Effect of the heart pass preparation on serum iron (SI) value in a rat rheumatoid arthritis related anemia model (compared with the model control group, *P<0.05, **P<0.01);

[0023] Figure 2 Effect of the heart pass preparation on transferrin saturation (TS) value in a rat rheumatoid arthritis related anemia model (compared with the model control group, *P<0.05, **P<0.01);

[0024] Figure 3 Effect of the heart pass preparation on serum hepcidin value in a rat rheumatoid arthritis related anemia model (compared with the model control group, *P<0.05, **P<0.01);

[0025] Figure 4 Effect of the heart pass preparation on hemoglobin (Hb) value of rats in a rat chronic kidney disease anemia model related anemia model. DETAILED DESCRIPTION

[0026] The above content of the application will be further described in detail through the specific embodiments in the form of examples. However, this should not be understood as the scope of the above subject matter of the application being limited to the following examples.

[0027] Experimental example 1: Effect of the heart pass preparation on a rat rheumatoid arthritis related anemia model

[0028] 1 Materials and methods

[0029] 1.1 Experimental animals and feeding

[0030] Healthy SPF level SD rats, half male and half female, experimental animal license number: SYXK (Lu) 20230023, body weight 200±20g. After 7 days of laboratory acclimation, the experiment was started.

[0031] 1.2 Model preparation

[0032] Rats were injected subcutaneously at the base of the tail with 0.1 ml of emulsified antigen of bovine collagen type II (2 mg / ml) mixed with complete Freund's adjuvant (1:1) to induce the model of arthritis anemia. On the 7th day of the experiment, the rats were boosted with the same dose of collagen mixed with incomplete Freund's adjuvant.

[0033] On the 21st day of the experiment, rats with an arthritis index (AI) of ≥8 points (on a scale of 0-16), Hb <100 g / L, and serum IL-6 >50 pg / mL were considered to have developed collagen-induced arthritis (CIA) anemia.

[0034] 1.3 Grouping and administration

[0035] The rats that met the criteria were randomly divided into the model control group, the positive control group, the low-dose Xintong preparation group, and the high-dose Xintong preparation group, with 10 rats in each group. The rats were administered according to the following table:

[0036]

[0037] The rats were continuously administered for 28 days according to the above method.

[0038] 2. Test indicators and methods

[0039] 2.1 Detection of key indicators of iron metabolism:

[0040] On the 21st day, the 35th day, and the 49th day of the experiment, blood was collected from the orbital venous plexus, and the hemoglobin (Hb) value was detected. Serum was obtained by centrifugation, and the serum iron (SI) value was determined using the ferrozine colorimetric method (kit purchased from Nanjing Jiancheng Biological Engineering Institute). The amount of unbound iron was determined after the serum combined with excess iron ions, and the TS was calculated:

[0041] Transferrin saturation (TS) = (SI / TIBC) x 100%.

[0042] Serum was also obtained, and the serum hepcidin was detected using the ELISA method (kit purchased from the United States R&D Systems, detection wavelength 450 nm, minimum detection limit 0.5 ng / mL).

[0043] 2.2 Detection of inflammatory factors and serum erythropoietin (EPO):

[0044] After the experiment, blood was collected from the orbital venous plexus, and serum was obtained by centrifugation. The TNF-α and IL-6 values were detected using the ELISA method. The TNF-α kit was purchased from BD Biosciences, and the IL-6 kit was purchased from Abeam. Serum was also obtained and detected using the ELISA method (kit purchased from the United States R&D Systems), with a detection range of 0.1-100 mIU / mL.

[0045] 2.3 Detection of indicators of arthritis severity

[0046] After the experiment, the four limbs of each rat were scored, with the score being:

[0047] Single limb 0-4 points: 0 = normal, 1 = local erythema / slight swelling, 2 = toe joint swelling, 3 = swelling below the ankle joint, 4 = whole limb swelling + deformity.

[0048] The circumference of the right hind limb 5 mm above the ankle joint was measured with a vernier caliper, and the average of three measurements was taken.

[0049] 2.4 Statistical processing: SPSS22.0 software was used for statistical analysis, and the experimental data were expressed in the form of "mean ± standard deviation

[0050] ". Comparison between multiple groups used one-way ANOVA, and P < 0.05 was considered statistically significant.

[0051] 3 Experimental results

[0052] 3.1 Hemoglobin (Hb) value results

[0053] Table 1 Hemoglobin (Hb) value

[0054]

[0055] Compared with the model control group, *P < 0.05, **P < 0.01.

[0056] It can be seen that the Hb of the high-dose Xintong group increased by 3.6 g / L compared with the positive control group at 49 days, although the difference was not significant (*P > 0.05), but the increase was 3.1%, indicating that the effect of Xintong preparation in improving anemia was comparable to that of the positive control.

[0057] 3.2 Serum iron (SI), transferrin saturation (TS), serum hepcidin value results

[0058] The results are shown in Figure 1 , Figure 2 , Figure 3 . It can be seen that the detection results at 49 days (28 days of administration) show that SI and TS are significantly improved compared with the model group, and Hepcidin value is significantly reduced, indicating that it can improve iron utilization by inhibiting hepcidin, and the results are better than those of the positive control group.

[0059] 3.3 Serum TNF-α, IL-6 value

[0060] The swelling degree of the high-dose Xintong group was reduced by 25.7% compared with the model group (*P<0.05), but was 20.0% higher than the positive control group (*P<0.05), indicating that the anti-inflammatory and detumescence effect of tofacitinib + EPO was stronger than that of Xintong preparation, and the high-dose Xintong could still significantly improve joint inflammation.

[0061] Table 2 Serum TNF-α, IL-6 values

[0062] Group TNF-α (pg / mL) IL-6 (pg / mL) Model control group 130.4±9.5 95.2±5.2 Positive control group 60.1±4.5** 40.1±3.4** Xintong high-dose group 50.6±3.2** 32.8±2.5** Xintong low-dose group 70.2±5.1** 50.1±3.1*

[0063] Compared with the model control group, *P<0.05, **P<0.01.

[0064] 3.4 Arthritis index and paw swelling degree

[0065] It was found that the score of the high-dose Xintong group was 3.2 after 28 days of administration, which was significantly lower than 9.3 of the model group and lower than that of the low-dose Xintong group and the positive control group.

[0066] The paw swelling degree values are shown in Table 3.

[0067] Table 3 Paw swelling degree values

[0068] Group Paw swelling degree (mm) Model control group 20.2±0.6 Positive control group 12.5±0.3** Xintong high-dose group 15.0±0.8* Xintong low-dose group 16.8±0.7*

[0069] Compared with the model control group, *P<0.05, **P<0.01.

[0070] As can be seen from Table 3, the swelling degree of the high-dose Xintong group was reduced after 28 days of administration, which was better than that of the positive control group and the low-dose group, and significantly better than that of the model control group, indicating that the anti-inflammatory and detumescence effect of Xintong preparation was weaker than that of the positive control, but the high dose still had a significant improvement effect.

[0071] Experimental Example 2 Effect of Xintong preparation on rat chronic kidney disease anemia model

[0072] 1 Materials and methods

[0073] 1.1 Experimental animals and feeding

[0074] Healthy SPF level SD rats, half male and half female, experimental animal license number: SYXK(Lu)20230023, body weight 200±20g. After 7 days of laboratory acclimation, the experiment was started.

[0075] 1.2 Model preparation

[0076] Preoperative fasting 12h, rats intraperitoneal injection of 3% sodium pentobarbital (30mg / kg) anesthesia, resection of the left kidney upper and lower end of 1 / 3, electrocoagulation hemostasis after suture; 1 week after the right kidney, postoperative intramuscular penicillin (40,000 U / kg) to prevent infection; 4 weeks after the operation detection of serum creatinine (SCr) ≥ 150 μmol / L and hemoglobin (Hb) <100g / L, for the success of the modeling.

[0077] 1.3 Grouping and administration

[0078] The standard rats were randomly divided into model control group, positive control group, heart Tong preparation group, 8 in each group.

[0079] According to the following table for administration:

[0080]

[0081]

[0082] According to the above method for continuous administration of 28 days.

[0083] 2 Test index and method

[0084] 2.1 Hemoglobin (Hb) detection and transferrin saturation (TSAT) detection

[0085] After the end of the experiment, the rat retro-orbital plexus blood was added to the EDTA anticoagulant tube to detect the Hb concentration (g / L), and the Sysmex XN-1000 automatic blood analyzer was used to detect the Hb concentration (g / L); another centrifugal serum, using ferrozine colorimetric method (kit from Nanjing Jiancheng Biological Engineering Institute) to determine the serum iron (SI) value, using double pyridine colorimetric method, serum combined with excess iron ions, the amount of unbound iron was measured, and TS was calculated:

[0086] Transferrin saturation (TS) = (SI / TIBC) x 100%;

[0087] Another serum was detected by ELISA method (kit from R&D Systems, USA), the detection wavelength was 450nm, the minimum detection limit was 0.5ng / mL, and the serum iron regulator (Hepcidin) was detected according to the operation instruction.

[0088] 2.2 Kidney tissue observation

[0089] After the end of the experiment, the rats were killed and the left kidney tissue was taken, fixed with 4% paraformaldehyde, dehydrated with ethanol, embedded in paraffin, sectioned, and observed.

[0090] 2.4 Statistical processing: SPSS22.0 software was used for statistical analysis, and the experimental data were expressed as "mean ± standard deviation The results are expressed in the form of mean ± SD. The comparison between groups was performed by one-way ANOVA, and P < 0.05 was considered statistically significant.

[0091] 3. Experimental results

[0092] 3.1 Serum iron (SI) and transferrin saturation (TS) are shown in Table 1.

[0093] After the experiment, the hemoglobin (Hb) values of the rats in each group are shown in Table 4, and the serum iron (SI) and transferrin saturation (TS) and Hepcidin values are shown in Table 4: Figure 4

[0094] Table 4 Serum iron (SI) and transferrin saturation (TS) and Hepcidin values

[0095] Group Hb (g / L) SI (μg / dL) TS (%) Hepcidin (ng / mL) Model control group 90.5±5.6 40.2±4 11.3±2.1 28.6±3.9 Positive control group 114.7±8.6** 78.5±6.5** 24.5±3.3** 21.5±2.1* Xintong preparation group 123.9±9.2** 82.8±7.1** 28.1±3.0** 19.5±2.5**

[0096] Compared with the model control group, *P < 0.05, **P < 0.01.

[0097] After the experiment, the hemoglobin (Hb), serum iron (SI), transferrin saturation (TS), and Hepcidin of the rats in each group were detected, and the results are shown in Table 4. The Hb, SI, and TS values of the Xintong preparation group were significantly increased, and Hepcidin decreased from 28 ng / mL to 19.5 ng / mL. The improvement of each index was better than that of the positive control group.

[0098] 3.2 Kidney tissue observation results

[0099] The experimental results showed that the kidneys of the model control group presented typical pathological changes of chronic kidney disease, including mesangial cell proliferation, basement membrane thickening, renal tubular epithelial cell swelling, and reduced blood vessel density. The pathological damage of the positive control group was reduced, while the kidney tissue repair of the Xintong preparation group was more significant, with mesangial proliferation relief, improved renal tubular structure, and obvious neovascularization.

[0100] Example 3: Study on the effect of Xintong preparation on a rat model of diabetes-related anemia

[0101] 1.1 Experimental animals and feeding

[0102] Healthy SPF grade SD rats, half male and half female, experimental animal license number: SYXK(Lu)20230023, body weight 200 ± 20 g. After 7 days of laboratory acclimation, the experiment began.

[0103] 1.2 Model preparation

[0104] The rats were fed with high-fat feed for 4 weeks, and STZ was injected intraperitoneally at 35 mg / kg. After 72 h, the fasting blood glucose was screened. ​

[0105] ≥11.1mmol / L and weight loss are the diabetes model. Continue to feed for 4 weeks, screening Hb <100g / L and SI <50μg / dL rats as diabetes anemia model.

[0106] 1.3 Grouping and administration

[0107] The qualified rats were randomly grouped (n=10 / group), respectively as model control group, positive control group, Xintong preparation group.

[0108] According to the following table, the administration was carried out:

[0109]

[0110] According to the above method, the administration was continuously carried out for 28 days.

[0111] 2 Test index and method

[0112] 2.1 Blood routine test

[0113] On the 1st day of administration and the last day of administration, the blood was taken from the jugular vein of rats, and the hemoglobin (Hb), red blood cell count (RBC) and hematocrit (HCT) of rats in each group were detected by automatic blood analyzer.

[0114] At the same time, on the 1st day of administration and the last day of administration, the blood was taken to determine the fasting blood glucose (FBG) value.

[0115] 2.2 Transferrin saturation (TSAT) and serum TNF-α, IL-6 value detection

[0116] After the experiment, the blood was taken from the jugular vein of rats, and the serum was centrifuged. The serum iron (SI) value was determined by ferrozine colorimetry (kit purchased from Nanjing Jiancheng Biological Engineering Institute), and the amount of unbound iron was determined after the serum combined with excess iron ions by dipyrine colorimetry, and the TS was calculated:

[0117] Transferrin saturation (TS) = (SI / TIBC) x 100%;

[0118] After the experiment, the serum hepcidin value and the serum TNF-α, IL-6 value were determined by ELISA method.

[0119] 3 Experimental results

[0120] 3.1, Blood routine test results:

[0121] On the 1st day of administration and the last day of administration, the blood was taken for detection, and the experimental results were as follows:

[0122] Table 5 Hemoglobin (Hb), red blood cell count (RBC), hematocrit (HCT) values of rats in each group

[0123]

[0124] Compared with the model control group, *P<0.05, **P<0.01.

[0125] After administration of Xintong preparation for 28 days, the Hb, RBC and HCT values were significantly increased, and the improvement of the three indicators was better than that of the positive control group, and was significantly better than that of the model control group (P<0.01), indicating that compared with the positive control group, Xintong preparation has a stronger promoting effect on erythropoiesis. The indicators of the model control group had no significant change before and after administration (P>0.05), indicating that the diabetic anemia model of the embodiment is stable.

[0126] Specifically, the fasting blood glucose results are as follows:

[0127] Table 6 FBG values of rats in each group

[0128]

[0129]

[0130] Compared with the model control group, *P<0.05, **P<0.01.

[0131] It can be seen that the fasting blood glucose reduction value of the Xintong preparation group is better than that of the positive control group, and the treatment effect on hyperglycemia in rats is better than that of the positive control drug.

[0132] The iron metabolism indicators are as follows:

[0133] Table 7 SI, TS% and hepcidin values of rats in each group

[0134] Group SI (μg / dL) TS (%) Hepcidin (ng / mL) Model control group 42.5±3.5 10.2±1.9 22.5±2.4 Positive control group 58.0±4.7** 16.2±2.4** 18.4±2.2* Xintong preparation group 70.7±5.2** 22.3±2.7** 12.4±1.4**

[0135] Compared with the model control group, *P<0.05, **P<0.01.

[0136] The serum iron and transferrin saturation values of the Xintong preparation group were significantly increased and were significantly higher than those of the model control group, and in particular, the serum hepcidin reduction value was much higher than that of the positive control group, indicating that the administration preparation Xintong of the application can promote iron utilization efficiency compared with the positive control group.

[0137] Table 8 TNF-α and IL-6 values of rats in each group

[0138] Group TNF-α (pg / mL) IL-6 (pg / mL) Model control group 90.2.±8.8 75.4±7.1 Positive control group 65.4±6.4** 55.3±5.8* Xintong preparation group 50.1±5.7** 40.1±4.9**

[0139] Compared with the model control group, *P<0.05, **P<0.01.

[0140] The above experimental results show that the serum inflammatory factors TNF-α and IL-6 of the Xintong preparation group are obviously decreased compared with the model control group, and the inflammation inhibition effect is significantly better than that of the positive control group.

[0141] In conclusion, the present application discloses a new use of the Xintong preparation in preparing a medicine for treating anemia, especially anemia caused by blood stasis and phlegm dampness. Experiments prove that the Xintong preparation can significantly improve the hemoglobin level of rheumatoid arthritis, chronic kidney disease and diabetic anemia model rats through multiple pathways such as inhibiting hepcidin, improving iron metabolism, reducing inflammatory response and promoting tissue repair, and the effect is better than that of traditional positive control drugs. At the same time, the Xintong preparation can improve blood glucose, kidney function and pathological damage, and provides a new treatment scheme for clinically refractory anemia.

Claims

1. Use of Xintong preparation in the preparation of a medicament for treating anemia, wherein the Xintong preparation is Xintong oral liquid, the anemia is secondary to rheumatoid arthritis, chronic kidney disease, diabetes; the Xintong preparation is prepared from 13 raw medicinal materials of Huangqi, Dangshen, Maidong, Heshouwu, Yinyanghe, Gegen, Dangui, Danshen, Zaokesheng, Haizao, Kelp, Muli, and Zhishi.

2. Use according to claim 1, characterized in that, The anemia is chronic anemia.

3. Use according to claim 1, characterized in that, Use of the Xintong preparation as the only drug in the preparation of a medicament for treating anemia.

4. Use according to claim 1, characterized in that, Use of the Xintong preparation in combination with a qi-tonifying and blood-producing drug in the preparation of a medicament for treating anemia.

5. Use according to claim 1, characterized in that, The Xintong preparation is prepared from the following raw medicinal materials in the weight parts:

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