Composition containing fucoidin and used for chronic nephritis and pharmaceutical composition

Abstract

The invention relates to the technical field of polysaccharide medicinal preparations, and particularly discloses a fucoidin-containing composition for chronic nephritis, which comprises the following raw materials: fucoidin, Chinese herbal medicine extract and auxiliary materials. The fucoidin is prepared by the following specific operations: (1) airing and pulping fresh kelp, adding into a tartaric acid solution, grinding for 1-2 hours, adding sodium bicarbonate, stirring, performing electric field pulse treatment for 2-4 minutes in the stirring process, adding a calcium chloride solution, stirring, centrifuging, performing solid-liquid separation, adding trichloroacetic acid into liquid, stirring, centrifuging, and collecting supernate to obtain first supernate; (2) adding a first part of absolute ethyl alcohol into the first supernatant, stirring, standing, centrifuging, and collecting the supernatant to obtain a second supernatant; and adding a second part of absolute ethyl alcohol into the second supernate, stirring, standing, centrifuging, collecting lower-layer precipitate, drying, grinding, adding into deionized water, centrifuging for 1-2 hours, collecting precipitate, washing, and freeze-drying.

Description

Technical Field

[0001] The present invention relates to the technical field of polysaccharide pharmaceutical preparations, and in particular to a fucoidan-containing composition for chronic nephritis, a pharmaceutical composition for chronic nephritis, and the use of the two in preparing drugs for treating chronic glomerulonephritis. Background Art

[0002] Chronic glomerulonephritis is referred to as chronic nephritis. Currently, only a few cases of chronic nephritis are caused by the development of acute nephritis, while the pathogenesis of most chronic nephritis is immune-mediated inflammation. Up to now, the clinical treatment of chronic nephritis is mainly to suppress inflammation through glucocorticoids, or to reduce kidney filtration pressure through antihypertensive drugs to relieve symptoms, which not only has limited effect, but also has many side effects.

[0003] my country is rich in kelp resources, and its output ranks first in the world. The results of modern molecular biology and medical research show that there is a polysaccharide between kelp cells, fucoidan, which has a wide range of biological activities, such as antiviral, anti-tumor, immune regulation, and anticoagulation.

[0004] Kelp fucoidan, also known as fucoidan sulfate and fucoidan, is a water-soluble sulfated heteropolysaccharide. With the in-depth study of the various activities of fucoidan, its application value continues to rise, resulting in growing demand. Currently, more and more research focuses on the extraction, purification and biological activity of fucoidan. Generally speaking, on the one hand, incomplete removal of impurities such as pigments and proteins during the pretreatment of raw materials may lead to fucoidan contamination; on the other hand, fucoidan undergoes physical, chemical and functional changes during the preparation process. Different methods often lead to differences in monosaccharide composition and weight-average molecular weight, resulting in different biological activities of polysaccharides.

[0005] At present, the extraction of fucoidan generally adopts hot water extraction method, but hot water extraction of polysaccharides is prone to cause polysaccharide degradation, thus affecting the biological activity and yield of polysaccharides. On the basis of hot water extraction, ultrasound-assisted extraction of fucoidan can improve the yield of polysaccharides, but the activity of fucoidan with different molecular weights is different. How to use the obtained fucoidan for chronic nephritis has extremely strong practical significance. Summary of the invention

[0006] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a fucoidan-containing composition for chronic nephritis, a pharmaceutical composition for chronic nephritis, and the use of the two in the preparation of drugs for treating chronic glomerulonephritis.

[0007] A fucoidan-containing composition for chronic nephritis, wherein the raw materials thereof include, by weight: 1-2 parts of fucoidan, 1-3 parts of Chinese herbal medicine extract, and 4-12 parts of auxiliary materials.

[0008] Preferably, fucoidan is prepared by the following specific operations:

[0009] (1) drying fresh kelp, beating the kelp, adding the tartaric acid solution to grind for 1-2 hours, adding sodium bicarbonate to stir, subjecting the mixture to electric field pulse treatment for 2-4 minutes during stirring, adding calcium chloride solution to stir, centrifuging, separating the solid from the liquid, adding trichloroacetic acid to the liquid to stir, centrifuging, and collecting the supernatant to obtain a first supernatant;

[0010] (2) adding the first portion of anhydrous ethanol to the first supernatant, stirring, letting stand, centrifuging, collecting the supernatant to obtain a second supernatant; then adding the second portion of anhydrous ethanol to the second supernatant, stirring, letting stand, centrifuging, collecting the lower layer of precipitate, drying, grinding, adding to deionized water, centrifuging for 1-2 hours, collecting the precipitate, washing, and freeze-drying.

[0011] Preferably, during the electric field pulse treatment in step (1), the electric field intensity is 60-80 MPa and the pulse frequency is 0.5-1 Hz.

[0012] Preferably, in step (1), the mass fraction of the tartaric acid solution is 40-60%, the mass fraction of the calcium chloride solution is 10-20%, and the mass ratio of the fresh kelp, the tartaric acid solution, the sodium bicarbonate, the calcium chloride solution, and the trichloroacetic acid is 10-20:30-60:5-15:1-5:1-2.

[0013] Preferably, in step (2), the mass ratio of the first supernatant, the first portion of anhydrous ethanol, and the second portion of anhydrous ethanol is 40-80:5-15:20-40.

[0014] Preferably, in step (2), the freeze-drying temperature is -20 to -30°C.

[0015] Preferably, the grinding pressure is 0.5-2.5 MPa, and the grinding speed is 1000-2000 r / min.

[0016] Preferably, the relative density of the Chinese herbal medicine extract is 1.1-1.2.

[0017] Preferably, the raw materials used in the Chinese herbal medicine extract include: astragalus, herbaceous euphorbia, and codonopsis pilosula; the mass ratio of astragalus, herbaceous euphorbia, and codonopsis pilosula is 10-20:5-15:1-10.

[0018] Preferably, the Chinese herbal medicine extract is prepared by the following specific steps: soaking Astragalus, Euphorbia pulex and Codonopsis pilosula in water for 10-20 hours, boiling over high heat and then decocting over low heat for 1-2 hours, decocting 1-3 times, combining the filtrate and concentrating.

[0019] A pharmaceutical composition for chronic nephritis comprises Shenkang injection and the above-mentioned fucoidan-containing composition for chronic nephritis.

[0020] Use of the above-mentioned fucoidan-containing composition for chronic nephritis or the above-mentioned pharmaceutical composition for chronic nephritis in the preparation of drugs for treating chronic glomerulonephritis.

[0021] The present invention adopts fucoidan and active ingredients of Chinese herbal medicine to obtain a composition for chronic nephritis, and adopts Shenkang injection to compound with the composition for chronic nephritis. It can not only fundamentally treat the problem of renal essence loss in chronic glomerulonephritis, but also has a stable and lasting kidney-tonifying effect, and can also effectively alleviate symptoms such as proteinuria and hematuria, and is suitable for the treatment of patients with chronic glomerulonephritis.

[0022] At the same time, the present invention adds fresh kelp to the tartaric acid solution after drying and pulping. Since tartaric acid has strong acidity, tartaric acid can fully enter the kelp structure during the grinding process, which can promote the better dissolution of fucoidan in the kelp on the one hand, and produce a large amount of gas under high-speed stirring in combination with subsequent sodium bicarbonate, which further enhances the dissolution degree of fucoidan. Under the comprehensive effect, not only the polysaccharide extraction rate is greatly improved, but also the polysaccharide molecular weight is narrow and the uniformity is good.

[0023] After the kelp is ground with tartaric acid, the tartaric acid enters the kelp structure, and then sodium bicarbonate is added to react. The research results show that the present invention can not only improve the polysaccharide extraction rate, but also change the structure of the polysaccharide and its physical and chemical properties, and optimize the biological activity.

[0024] The above extraction process does not require heating, so that the fucoidan in the solution does not react with proteins, amino acids and other components in the solution, so that the obtained fucoidan has good immune activity and less component loss.

[0025] The experimental results show that the present invention can not only reduce or lower urine protein, urine red blood cells, blood creatinine, blood urea nitrogen, improve renal function, but also has an excellent control effect on the development of lesions, and its total effective rate can reach more than 96%, and there is no obvious toxic and side effect. The present invention is simple and easy to obtain, safe, and has no toxic and side effects, and can avoid further damage to the kidneys caused by taking drugs. At the same time, the method is simple and convenient for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a comparison chart of 24-hour urine protein quantification before and after treatment of Example 5 group, comparison group 1 group, comparison group 2 group, comparison group 3 group and control group.

[0027] Figure 2 The following is a comparison chart of urine red blood cells before and after treatment of Example 5 group, comparison group 1 group, comparison group 2 group, comparison group 3 group and control group.

[0028] Figure 3It is a comparison chart of the decrease rates of serum creatinine and blood urea nitrogen before and after treatment in Example 5 group, comparison group 1 group, comparison group 2 group, comparison group 3 group and control group. DETAILED DESCRIPTION

[0029] The present invention will be further explained below in conjunction with specific embodiments.

[0030] Example 1

[0031] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 1kg of fucoidan, 1kg of Chinese herbal medicine extract, and 4kg of auxiliary materials.

[0032] Fucoidan is prepared by the following specific operations:

[0033] (1) Dry 10 kg of fresh kelp, beat it, add it to 30 kg of 40% tartaric acid solution, grind it for 1 hour, the grinding pressure is 0.5 MPa, the grinding speed is 1000 r / min, add 5 kg of sodium bicarbonate, stir it at 1000 r / min for 10 minutes, and treat it with electric field pulse for 2 minutes during the stirring process, the electric field strength is 60 MPa, and the pulse frequency is 0.5 Hz; add 1 kg of 10% calcium chloride solution, stir it at 500 r / min for 1 hour, centrifuge it, separate the solid and liquid, add 1 kg of trichloroacetic acid to the liquid, stir it at 1000 r / min for 10 minutes, centrifuge it, and collect the supernatant to obtain a first supernatant;

[0034] (2) Add 5 kg of anhydrous ethanol to 40 kg of the first supernatant, stir at a speed of 100 r / min for 1 hour, let stand for 1 day, centrifuge, collect the supernatant to obtain a second supernatant; add 20 kg of anhydrous ethanol to the second supernatant, stir at a speed of 1000 r / min for 1 hour, let stand for 1 day, centrifuge, collect the lower precipitate, dry at a temperature of 30°C, grind and add to 20 kg of deionized water, centrifuge at a speed of 5000 r / min for 1 hour, collect the precipitate, wash it twice with a 90% ethanol solution and ether respectively, freeze-dry it at a temperature of -20°C, and crush it to obtain fucoidan.

[0035] The Chinese herbal medicine extract is prepared by the following specific steps: 10 kg of astragalus, 5 kg of euphorbia pulex and 1 kg of codonopsis pilosula are added into 4.5 times the weight of water, soaked for 10 hours, boiled over high heat and then simmered over low heat for 60 minutes, decocted three times and the filtrate was combined and concentrated at a temperature of 50°C to a relative density of 1.1.

[0036] Example 2

[0037] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 2kg of fucoidan, 3kg of Chinese herbal medicine extract, and 12kg of auxiliary materials.

[0038] Fucoidan is prepared by the following specific operations:

[0039] (1) Dry 20 kg of fresh kelp, beat it, add it to 60 kg of 60% tartaric acid solution, grind it for 2 h, the grinding pressure is 2.5 MPa, the grinding speed is 2000 r / min, add 15 kg of sodium bicarbonate, stir it at 2000 r / min for 30 min, and treat it with electric field pulse for 4 min during stirring, the electric field strength is 80 MPa, and the pulse frequency is 1 Hz; add 5 kg of 20% calcium chloride solution, stir it at 1000 r / min for 3 h, centrifuge it, separate the solid and liquid, add 2 kg of trichloroacetic acid to the liquid, stir it at 4000 r / min for 20 min, centrifuge it, and collect the supernatant to obtain a first supernatant;

[0040] (2) Add 15 kg of anhydrous ethanol to 80 kg of the first supernatant, stir at a speed of 500 r / min for 2 h, let stand for 2 days, centrifuge, collect the supernatant to obtain a second supernatant; add 40 kg of anhydrous ethanol to the second supernatant, stir at a speed of 5000 r / min for 2 h, let stand for 2 days, centrifuge, collect the lower precipitate, dry at a temperature of 40°C, grind and add to 40 kg of deionized water, centrifuge at a speed of 15000 r / min for 2 h, collect the precipitate, wash it with a 95% ethanol solution and ether for 4 times respectively, freeze-dry it at a temperature of -30°C, and crush it to obtain fucoidan.

[0041] The Chinese herbal medicine extract is prepared by the following specific steps: 20 kg of astragalus, 15 kg of euphorbia pulex and 10 kg of codonopsis pilosula are added into 4.5 times of their weight of water, soaked for 20 hours, boiled over high heat and then simmered over low heat for 120 minutes, decocted three times and the filtrate was combined and concentrated at a temperature of 60°C to a relative density of 1.2.

[0042] Example 3

[0043] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 1.3 kg of fucoidan, 2.5 kg of Chinese herbal medicine extract, and 6 kg of auxiliary materials.

[0044] Fucoidan is prepared by the following specific operations:

[0045] (1) Dry 12 kg of fresh kelp, beat it, add it to 50 kg of 45% tartaric acid solution, grind it for 100 min, the grinding pressure is 1 MPa, the grinding speed is 1800 r / min, add 8 kg of sodium bicarbonate, stir it at 1700 r / min for 15 min, and treat it with electric field pulse for 2.5 min during stirring, the electric field strength is 75 MPa, and the pulse frequency is 0.6 Hz; add 2 kg of 17% calcium chloride solution, stir it at 600 r / min for 2.5 h, centrifuge it, separate the solid and liquid, add 1.8 kg of trichloroacetic acid to the liquid, stir it at 2000 r / min for 18 min, centrifuge it, and collect the supernatant to obtain a first supernatant;

[0046] (2) Add 8 kg of anhydrous ethanol to 50 kg of the first supernatant, stir at 400 r / min for 80 min, let stand for 1.5 days, centrifuge, collect the supernatant to obtain a second supernatant; add 35 kg of anhydrous ethanol to the second supernatant, stir at 2000 r / min for 100 min, let stand for 1.5 days, centrifuge, collect the lower precipitate, dry at 33°C, grind and add to 35 kg of deionized water, centrifuge at 8000 r / min for 100 min, collect the precipitate, wash it three times with 92% ethanol solution and ether respectively, freeze-dry it at -28°C, and crush it to obtain fucoidan.

[0047] The Chinese herbal medicine extract is prepared by the following specific steps: 12 kg of astragalus, 12 kg of euphorbia pulex and 3 kg of codonopsis pilosula are added into 4.5 times of their weight of water, soaked for 18 hours, boiled over high heat and then simmered over low heat for 80 minutes, decocted three times and the filtrate was combined and concentrated at a temperature of 58°C to a relative density of 1.15.

[0048] Example 4

[0049] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 1.7 kg of fucoidan, 1.5 kg of Chinese herbal medicine extract, and 10 kg of auxiliary materials.

[0050] Fucoidan is prepared by the following specific operations:

[0051] (1) drying 18 kg of fresh kelp, beating the kelp, adding 40 kg of 55% tartaric acid solution and grinding for 80 min, grinding pressure of 2 MPa, grinding speed of 1200 r / min, adding 12 kg of sodium bicarbonate, stirring at 1300 r / min for 25 min, and subjecting the mixture to electric field pulse treatment for 3.5 min during stirring, with the electric field strength of 65 MPa and the pulse frequency of 0.9 Hz; adding 4 kg of 13% calcium chloride solution, stirring at 800 r / min for 1.5 h, centrifuging, solid-liquid separation, adding 1.2 kg of trichloroacetic acid to the liquid, stirring at 3000 r / min for 12 min, centrifuging, and collecting the supernatant to obtain a first supernatant;

[0052] (2) Add 12 kg of anhydrous ethanol to 70 kg of the first supernatant, stir at a speed of 200 r / min for 100 min, let stand for 1.5 days, centrifuge, collect the supernatant to obtain a second supernatant; add 25 kg of anhydrous ethanol to the second supernatant, stir at a speed of 4000 r / min for 80 min, let stand for 1.5 days, centrifuge, collect the lower layer of precipitate, dry at a temperature of 37°C, grind and add to 25 kg of deionized water, centrifuge at a speed of 12000 r / min for 80 min, collect the precipitate, wash it three times with a mass fraction of 94% ethanol solution and ether respectively, freeze-dry it at a temperature of -22°C, and crush it to obtain fucoidan.

[0053] The Chinese herbal medicine extract is prepared by the following specific steps: 18 kg of astragalus, 8 kg of euphorbia pulex and 7 kg of codonopsis pilosula are added into 4.5 times of their weight of water, soaked for 12 hours, boiled over high heat and then simmered over low heat for 100 minutes, decocted three times and the filtrate was combined and concentrated at a temperature of 52°C to a relative density of 1.15.

[0054] Example 5

[0055] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 1.5 kg of fucoidan, 2 kg of Chinese herbal medicine extract, and 8 kg of auxiliary materials.

[0056] Fucoidan is prepared by the following specific operations:

[0057] (1) drying 15 kg of fresh kelp, beating the kelp, adding 45 kg of 50% tartaric acid solution and grinding for 90 min, grinding pressure of 1.5 MPa, grinding speed of 1500 r / min, adding 10 kg of sodium bicarbonate, stirring at 1500 r / min for 20 min, and subjecting the mixture to electric field pulse treatment for 3 min during stirring, with the electric field strength of 70 MPa and the pulse frequency of 0.75 Hz; adding 3 kg of 15% calcium chloride solution, stirring at 700 r / min for 2 h, centrifuging, solid-liquid separation, adding 1.5 kg of trichloroacetic acid to the liquid, stirring at 2500 r / min for 15 min, centrifuging, and collecting the supernatant to obtain a first supernatant;

[0058] (2) Add 10 kg of anhydrous ethanol to 60 kg of the first supernatant, stir at a speed of 300 r / min for 90 min, let stand for 1.5 days, centrifuge, collect the supernatant to obtain a second supernatant; add 30 kg of anhydrous ethanol to the second supernatant, stir at a speed of 3000 r / min for 90 min, let stand for 1.5 days, centrifuge, collect the lower precipitate, dry at a temperature of 35°C, grind and add to 30 kg of deionized water, centrifuge at a speed of 10000 r / min for 90 min, collect the precipitate, wash it three times with a mass fraction of 93% ethanol solution and ether respectively, freeze-dry it at a temperature of -25°C, and crush it to obtain fucoidan.

[0059] The Chinese herbal medicine extract is prepared by the following specific steps: 15 kg of astragalus, 10 kg of euphorbia pulex and 5 kg of codonopsis pilosula are added into 4.5 times the weight of water, soaked for 15 hours, boiled over high heat and then simmered over low heat for 90 minutes, decocted three times and the filtrate was combined, and concentrated at a temperature of 52-58°C to a relative density of 1.15.

[0060] Comparative Example 1

[0061] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 1.5 kg of fucoidan, 2 kg of the Chinese herbal medicine extract obtained in Example 5, and 8 kg of auxiliary materials.

[0062] Fucoidan is prepared by the following specific operations:

[0063] (1) Dry 15 kg of fresh kelp, beat it, add it to 45 kg of 50% tartaric acid solution, grind it for 90 min, the grinding pressure is 1.5 MPa, the grinding speed is 1500 r / min, continue stirring for 20 min, and treat it with electric field pulse for 3 min during stirring, the electric field strength is 70 MPa, and the pulse frequency is 0.75 Hz; add 3 kg of 15% calcium chloride solution, stir it at a speed of 700 r / min for 2 h, centrifuge it, separate the solid and liquid, add 1.5 kg of trichloroacetic acid to the liquid, stir it at a speed of 2500 r / min for 15 min, centrifuge it, and collect the supernatant to obtain a first supernatant;

[0064] S2. Add 10 kg of anhydrous ethanol to 60 kg of the first supernatant, stir at a speed of 300 r / min for 90 min, let stand for 1.5 days, centrifuge, collect the supernatant to obtain a second supernatant; add 30 kg of anhydrous ethanol to the second supernatant, stir at a speed of 3000 r / min for 90 min, let stand for 1.5 days, centrifuge, collect the lower precipitate, dry at a temperature of 35°C, grind and add to 30 kg of deionized water, centrifuge at a speed of 10000 r / min for 90 min, collect the precipitate, wash it three times with a mass fraction of 93% ethanol solution and ether respectively, freeze-dry it at a temperature of -25°C, and crush it to obtain fucoidan.

[0065] The fucoidans obtained in Example 5 and Comparative Example 1 were placed in sample bottles, 10 mL of 1 mol / L hydrochloric acid solution was added, and hydrolyzed at 30°C for 4 h. After the hydrolysis was completed, the filtrate was filtered and stored, and the sulfate content was determined according to the sulfate content determination method. The sulfate content of the fucoidan obtained in Example 5 was 20.74%, while the sulfate content of the fucoidan obtained in Comparative Example 1 was 18.26, confirming that the main components of both were indeed fucoidan.

[0066] The yields of fucoidan obtained in Example 5 and Comparative Example 1 were measured to be 13.75% and 9.21%, respectively, confirming that the combination of tartaric acid and sodium bicarbonate in the present application can enhance the dissolution degree of fucoidan and improve the extraction rate of fucoidan.

[0067] The fucoidan obtained in Example 5 was prepared into a 0.5 mg / mL solution with a mobile phase, and the molecular weight and molecular weight distribution of the polysaccharide were analyzed using a Wyatt GPC / SEC-MALS high-performance exclusion chromatography coupled with a multi-angle laser light scattering system (HPSEC-MALLS). The detectors connected in series with the HPSEC include a multi-angle laser light scattering detector, a viscosity detector, and a parallax detector. The Model 1500 HPLC pump is connected to two analytical columns: SB-806HQ and SB-804HQ (ShodexOHpak, 8mm×300mm, Showa Denko K.K., Japan). The temperature of the chromatographic column detector was maintained at 45°C, and the mobile phase was a 0.02% NaNO3 aqueous solution passing through a 0.45μm membrane, with a flow rate of 0.6mL / min. Finally, the data were collected and analyzed using ASTRA6.1software.

[0068] The data show that the peak shape of the fucoidan obtained in Example 5 is symmetrical and uniform, and its molecular weight dispersion (PDI value) is only 1.35, indicating that the combination of tartaric acid and sodium bicarbonate in this application can not only enhance the dissolution degree of fucoidan, but also make the obtained fucoidan have a narrower molecular weight distribution and good uniformity.

[0069] Comparative Example 2

[0070] A fucoidan-containing composition for chronic nephritis, the raw materials of which include: 2 kg of the Chinese herbal medicine extract obtained in Example 5 and 8 kg of auxiliary materials.

[0071] A total of 151 patients diagnosed with chronic glomerulonephritis in the outpatient department of nephrology of our hospital were selected (inclusion criteria: (1) meeting the diagnosis of chronic glomerulonephritis; (2) aged between 18 and 70 years old, regardless of gender; (3) 0.5 g ≤ 24 h urine protein ≤ 2.0 g; (4) excluding other chronic diseases, such as diabetes; (5) the patient or his / her guardian agreed and signed an informed consent form). They were aged 35-51 years old with an average age of 44.2 years old and were randomly divided into 4 groups, namely control group (30 people), example 5 group (30 people), comparison group 1 (31 people), comparison group 2 (30 people) and comparison group 3 (30 people).

[0072] Control group: On the basis of general treatment (i.e., restricting diet, preventing infection, and avoiding fatigue), patients were given losartan potassium tablets 50 mg (Hangzhou Merck Pharmaceutical Co., Ltd., National Medicine Approval No.: J20130148), once a day, 1 tablet each time, orally in the morning.

[0073] Group 5 of Example: On the basis of general treatment, the patients were given 50 mL of Shenkang injection (Xi'an Century Shengkang Pharmaceutical Co., Ltd., National Medicine Standard No. Z20040110) per day (added to 150 mL of 10% glucose injection, intravenous drip), and at the same time, the fucoidan-containing composition for chronic nephritis obtained in Example 5 was given (0.5 g / pill, 2 pills / time, 3 times / day, orally after meals).

[0074] Comparative group 1: On the basis of general treatment, the fucoidan-containing composition for chronic nephritis obtained in Example 5 was administered (0.5 g / capsule, 2 capsules / time, 3 times / day, orally after meals).

[0075] Comparison Group 2: On the basis of general treatment, the subjects were given Shenkang Injection (Xi'an Century Shengkang Pharmaceutical Co., Ltd., National Medicine Standard No. Z20040110) 50 mL per day (added to 150 mL 10% glucose injection, intravenous drip), and at the same time, the fucoidan-containing composition for chronic nephritis obtained in Comparative Example 1 was given (2 capsules / time, 3 times / day, orally after meals).

[0076] Comparison Group 3: On the basis of general treatment, the subjects were given Shenkang Injection (Xi'an Century Shengkang Pharmaceutical Co., Ltd., National Medicine Standard No. Z20040110) 50 mL per day (added to 150 mL 10% glucose injection, intravenous drip), and at the same time, the fucoidan-containing composition for chronic nephritis obtained in Comparative Example 2 was given (2 capsules / time, 3 times / day, orally after meals).

[0077] After 4 weeks of treatment, the test indicators of the patients before and after treatment were compared.

[0078] Before treatment, there was no significant difference in 24h-Upro among the five groups of patients (P>0.05). After treatment, 24h-Upro of the five groups of patients decreased compared with that before treatment, and the difference was statistically significant (P<0.05).

[0079] Compared with the control group, the reduction degree of 24h-Upro in Example 5, Comparison Group 1, Comparison Group 2 and Comparison Group 3 was significantly greater than that in the control group (P < 0.05), and the difference was statistically significant. The reduction degree of 24h-Upro in Example 5 was the highest, reaching 89.76%, which was better than the other groups.

[0080] 2. Compare the urine red blood cells of each group of patients before and after treatment. Figure 2 shown.

[0081] Before treatment, there was no significant difference in urine red blood cells among the five groups of patients (P>0.05). After treatment, urine red blood cells in the five groups of patients decreased compared with those before treatment, and the difference was statistically significant (P<0.05).

[0082] Compared with the control group, the reduction degree of urine red blood cells in Example 5, Comparison Group 1, Comparison Group 2 and Comparison Group 3 was significantly greater than that in the control group (P < 0.05), and the difference was statistically significant. The reduction degree of urine red blood cells in Example 5 was the highest, reaching 66.59%, which was better than the other groups.

[0083] 3. Compare the serum creatinine (SCr) and blood urea nitrogen (BUN) of each group of patients before and after treatment. Figure 3 shown.

[0084] After treatment, the decrease rates of serum creatinine and blood urea nitrogen in the control group were extremely low, indicating that there was no significant change in this group before and after treatment (P>0.05). However, the change rates of serum creatinine and blood urea nitrogen in Example 5 group were greatly changed compared with the control group (P<0.05), and the difference was statistically significant. This indicates that the pharmaceutical composition obtained in this application can stably reduce serum creatinine and urea nitrogen in patients with chronic nephritis and effectively protect renal function.

[0085] Efficacy criteria:

[0086] (1) Complete remission: Clinical symptoms and positive signs completely disappear, laboratory tests show that urine protein and red blood cells continue to turn negative, 24-hour urine protein is less than 0.2 g, and renal function recovers or remains normal for more than 3 months.

[0087] (2) Basic remission: Clinical symptoms and positive signs have basically disappeared, urine protein and urine red blood cells have decreased by ≥50% compared with before treatment, renal function has recovered or remained normal, or serum creatinine (SCr) has not increased significantly compared with before treatment and is within the normal range, or SCr has increased by <50% and lasted for more than 3 months.

[0088] (3) Effective: Clinical symptoms and positive signs are significantly improved, urine protein and / or urine red blood cells decrease by ≥25% compared with before treatment, renal function improvement lasts for more than 3 months, and SCr increases by <100% compared with the baseline value.

[0089] (4) Ineffective: No changes in clinical manifestations and laboratory tests.

[0090] According to the above efficacy judgment criteria, the efficacy of each group was statistically analyzed as shown in the following table:

[0091]

[0092] The total effective rate of Example 5 group was higher than that of the other groups, and the difference was statistically significant (P < 0.05), indicating that the present application has excellent therapeutic effect in treating chronic nephritis using fucoidan, Chinese herbal medicine extract and Shenkang injection.

[0093] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A composition for chronic nephritis containing fucoidan, characterized in that: The raw materials include: fucoidan, Chinese herbal medicine extract and auxiliary materials, and the mass ratio of fucoidan, Chinese herbal medicine extract and auxiliary materials is 1-2:1-3:4-12.

2. The fucoidan-containing composition for chronic nephritis according to claim 1, characterized in that: Fucoidan is prepared by the following specific operations: (1) drying fresh kelp, beating the kelp, adding the tartaric acid solution to grind for 1-2 hours, adding sodium bicarbonate to stir, subjecting the mixture to electric field pulse treatment for 2-4 minutes during stirring, adding calcium chloride solution to stir, centrifuging, separating the solid from the liquid, adding trichloroacetic acid to the liquid to stir, centrifuging, and collecting the supernatant to obtain a first supernatant; (2) adding the first portion of anhydrous ethanol to the first supernatant, stirring, letting stand, centrifuging, collecting the supernatant to obtain a second supernatant; then adding the second portion of anhydrous ethanol to the second supernatant, stirring, letting stand, centrifuging, collecting the lower layer of precipitate, drying, grinding, adding to deionized water, centrifuging for 1-2 hours, collecting the precipitate, washing, and freeze-drying.

3. The fucoidan-containing composition for chronic nephritis according to claim 2, characterized in that: During the electric field pulse treatment in step (1), the electric field strength is 60-80 MPa and the pulse frequency is 0.5-1 Hz.

4. The fucoidan-containing composition for chronic nephritis according to claim 2, characterized in that: In step (1), the mass fraction of the tartaric acid solution is 40-60%, the mass fraction of the calcium chloride solution is 10-20%, and the mass ratio of the fresh kelp, the tartaric acid solution, the sodium bicarbonate and the calcium chloride solution is 10-20:30-60:5-15:1-5.

5. The fucoidan-containing composition for chronic nephritis according to claim 2, characterized in that: In step (2), the mass ratio of the preformed liquid, trichloroacetic acid, the first portion of anhydrous ethanol, and the second portion of anhydrous ethanol is 40-80:1-2:5-15:20-40.

6. The fucoidan-containing composition for chronic nephritis according to claim 1, characterized in that: The relative density of Chinese herbal medicine extract is 1.1-1.

2.

7. The fucoidan-containing composition for chronic nephritis according to claim 1, characterized in that: The raw materials used in the Chinese herbal medicine extract include: astragalus, herba euphorbiae and codonopsis pilosula; the mass ratio of astragalus, herba euphorbiae and codonopsis pilosula is 10-20:5-15:1-10.

8. The fucoidan-containing composition for chronic nephritis according to claim 1, characterized in that: The Chinese herbal medicine extract is prepared by the following specific steps: soaking astragalus, herba euphorbiae and codonopsis pilosula in water for 10-20 hours, boiling over high heat and then decocting over low heat for 1-2 hours, decocting 1-3 times, combining the filtrate and concentrating.

9. A pharmaceutical composition for chronic nephritis, characterized in that: The invention comprises Shenkang injection and a composition for chronic nephritis containing fucoidan as claimed in any one of claims 1 to 8.

10. Use of the fucoidan-containing composition for chronic nephritis according to any one of claims 1 to 8, or the pharmaceutical composition for chronic nephritis according to claim 9 in the preparation of a drug for treating chronic glomerulonephritis.

Images