Rutin composition for treating arthritis and preparation method thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a rutin composition for treating arthritis and a preparation method thereof. The rutin composition for treating arthritis is prepared from the following active ingredients in parts by weight: 20 to 25 parts of rutin, 10 to 18 parts of typha orientalis glucoside and 8 to 15 parts of a resveratrol derivative. The rutin in the composition has remarkable anti-inflammatory and anti-oxidation effects and can relieve joint inflammation reaction, the tympanoside has analgesic and anti-inflammatory characteristics, the rutagenol derivative has anti-inflammatory and cartilage protection effects, the tympanoside and the rutagenol derivative have a synergistic effect with the rutin, the treatment effect of the composition is enhanced, and the three active ingredients are matched for use, so that the curative effect of the composition is improved. And a new scheme is provided for arthritis treatment.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a rutin composition for treating arthritis and a preparation method thereof. BACKGROUND

[0002] Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease characterized by synovial inflammation of the joints, which is characterized by chronic inflammatory joint lesions and accompanied by multiple systemic involvement. Without systematic treatment, it can be repeated for many years, eventually leading to joint deformity, loss of function, and causing great pain to patients.

[0003] At present, the commonly used arthritis treatment drugs in clinical practice mainly include non-steroidal anti-inflammatory drugs (NSAIDs), glucocorticoids and disease-modifying antirheumatic drugs (DMARDs), etc. NSAIDs are the first choice for the treatment of RA, including salicylic acid, traditional non-steroidal anti-inflammatory drugs and selective cyclooxygenase-2 (COX-2) inhibitors. However, long-term use of these drugs is often accompanied by serious side effects, such as gastrointestinal damage, liver and kidney function damage, and increased cardiovascular risk. Therefore, researchers are looking for treatment opportunities from various angles, and Chinese herbal medicines are increasingly valued for their low toxicity, multi-target treatment and low cost, becoming a new hotspot for the development of RA treatment drugs.

[0004] Rutin is a natural flavonoid compound, widely exists in sophora, buckwheat, citrus and other plants, and has significant anti-inflammatory, antioxidant and immunomodulatory effects. Studies have shown that rutin can inhibit the production of inflammatory factors such as TNF-α, IL-1β and IL-6, reduce synovial inflammation, and protect chondrocytes. Patent CN118217221A discloses a preparation method of rutin-loaded black phosphorus nanosheet composite hydrogel and its anti-arthritic application. However, single rutin preparation has limitations such as low bioavailability and single target, which affects its clinical efficacy. Typhaneoside is an effective component extracted from traditional Chinese medicine Typha angustata. Modern pharmacological studies have confirmed that it has analgesic, anti-inflammatory and immunomodulatory effects. Resveratrol is a natural polyphenolic compound, widely exists in grapes, giant knotweed and other plants, and has been confirmed to have anti-inflammatory, antioxidant, immunomodulatory and cartilage protection effects. It can inhibit the release of TNF-α, IL-1β and IL-6 and other pro-inflammatory factors through the inhibition of NF-κB and MAPK inflammatory signaling pathways, but has defects such as low bioavailability and poor stability.

[0005] Although there are reports on the use of rutin for the treatment of arthritis in the prior art, the combination of rutin, typhaneoside and resveratrol derivatives has not been disclosed. SUMMARY

[0006] In order to overcome the deficiencies of the prior art, the primary object of the present application is to provide a rutin composition for treating arthritis, wherein the rutin has significant anti-inflammatory and antioxidant effects, can reduce the inflammatory response of arthritis, the typhaneoside has analgesic and anti-inflammatory properties, the orobanchyl alcohol derivative has anti-inflammatory and cartilage protection effects, and the two have a synergistic effect with the rutin, thereby enhancing the therapeutic effect of the composition, and the combination of the three active ingredients provides a new scheme for treating arthritis.

[0007] Another object of the present application is to provide a preparation method of the rutin composition for treating arthritis.

[0008] The object of the present application is achieved by the following technical solutions. A rutin composition for treating arthritis, comprising the following active ingredients in parts by weight: 20-25 parts of rutin, 10-18 parts of typhaneoside, and 8-15 parts of orobanchyl alcohol derivative; the orobanchyl alcohol derivative has the following structural formula: .

[0009] Further, the orobanchyl alcohol derivative is prepared by the following preparation process: (1) 2-indolone and furfural are added to a solvent, and a catalyst is added, and reflux reaction is carried out; after the reaction is completed, filtration, washing, purification are carried out to obtain intermediate 1; (2) Resveratrol is added to water, and sodium hydroxide, 1,2-dichloroethane and tetrabutylammonium bromide are added, and stirring reaction is carried out; after the reaction solution is extracted, the organic phases are combined, and the organic phase is washed with water, dried, filtered, concentrated, and purified to obtain intermediate 2; (3) Intermediate 2 is added to a solvent, and intermediate 1 and K2CO3 are added, and stirring reaction is carried out; after the reaction solution is extracted, the organic phases are combined, and the organic phase is washed with water, dried, filtered, concentrated, and purified to obtain the orobanchyl alcohol derivative.

[0010] Further, in step (1), the amount ratio of 2-indolone, furfural, catalyst, and solvent is 10 mmol: 11-12 mmol: 0.1-0.5 mmol: 15-20 mL, the catalyst is piperidine, and the solvent is ethanol.

[0011] Further, in step (1), the reflux reaction time is 4-5 h.

[0012] Further, in step (2), the amount ratio of resveratrol, sodium hydroxide, 1,2-dichloroethane, tetrabutylammonium bromide, and water is 20 mmol: 75-80 mmol: 180-200 mmol: 2-2.2 mmol: 15-20 mL.

[0013] Further, in step (2), the reaction temperature is 75-85℃, and the reaction time is 12-14 h.

[0014] Further, in step (3), the ratio of the intermediate 2, the intermediate 1, K2CO3 and the solvent is 10 mmol: 50-60 mmol: 20-25 mmol: 80-100 mL; and the solvent is DMF.

[0015] Further, in step (3), the reaction temperature is 50-60℃, and the reaction time is 24-30 h.

[0016] The present application also provides a preparation method of the rutin composition for treating arthritis, which comprises the following steps: mixing the raw materials according to the weight parts to obtain the rutin composition.

[0017] The present application has the following effects compared with the prior art: 1. The rutin in the composition has significant anti-inflammatory and antioxidant effects, which can reduce the inflammatory response of arthritis. The typhaneoside has analgesic and anti-inflammatory properties, and the alisol B 11 has anti-inflammatory and cartilage protection effects. The two components have a synergistic effect with the rutin, which enhances the therapeutic effect of the composition, improves the swelling of the foot and the inflammation of arthritis caused by arthritis, and also reduces the serum collagen-specific antibody level of the arthritis mouse. The three active ingredients are used in combination, which has a good therapeutic effect on collagen-induced rheumatoid arthritis in mice. The composition provides a new scheme for treating arthritis.

[0018] 2. The rigid indole ketone-furan heterocyclic structure is introduced at the phenolic hydroxyl group of alisol B 11, which enhances the stability of the whole molecule, and the expansion of the conjugated system can effectively disperse the unpaired electrons of the free radicals, thereby reducing the damage of oxidative stress to the molecule. Compared with alisol B 11, the derivative after structural modification has stronger anti-inflammatory effect and performs better in anti-arthritis. DETAILED DESCRIPTION

[0019] The technical solutions of the present application are further described below in combination with the specific embodiments. However, those skilled in the art should understand that the following examples are only used to illustrate the present application, and should not be regarded as limiting the present application. The specific conditions not mentioned in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, such as those not specifically mentioned, are conventional products obtained through commercial channels.

[0020] Example 1 A rutin composition for treating arthritis, which comprises the following active ingredients in weight parts: rutin (CAS No.: 153-18-4) 22 parts, typhaneoside (CAS No.: 104472-68-6) 15 parts, and alisol B 11 derivative 12 parts.

[0021] The resveratrol derivative was prepared by the following process: (1) 2-Indolone and furfural were added to ethanol, and then piperidine was added as a catalyst. The ratio of 2-indolone, furfural, piperidine, and ethanol was 10 mmol: 11.5 mmol: 0.3 mmol: 18 mL. The mixture was refluxed for 4.5 h. After the reaction was completed, the mixture was filtered, the product was collected and washed with ethanol, and then purified by silica gel column chromatography (chloroform / methanol = 9:1) to obtain intermediate 1 (yield 86.4%). 1 H NMR (C 13 H9NO2, 400 MHz, d6-DMSO) δ 10.89 (s, 1H, -NH), 8.75-8.73(d, 1H, Ar-H), 8.15-8.13(d, 1H, Ar-H), 7.67-7.65 (d, 1H, Ar-H), 7.55 (s, HRMS (ESI) + ): [M+H] + The calculation yields 212.06, and the result is 216.07.

[0022] (2) Resveratrol was added to water, followed by sodium hydroxide, 1,2-dichloroethane and tetrabutylammonium bromide. The ratio of resveratrol, sodium hydroxide, 1,2-dichloroethane, tetrabutylammonium bromide and water was 20 mmol:78 mmol:190 mmol:2.1 mmol:18 mL. The mixture was stirred at 80 °C for 13 h. The reaction solution was cooled to room temperature, diluted with water and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined, washed with water, dried, filtered, concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 95 / 5) to obtain intermediate 2 (yield 80.5%). 1 H NMR (C 20 H 21Cl3O3, 400 MHz, d6-DMSO) δ7.82-7.79 (d, 2H, Ar-H), 7.08-7.05(d, 2H, Ar-H), 6.92-6.89 (d, 1H, CH), 6.84(s, 2H, Ar-H), 6.83-6.82 (d, 1H, CH), 6.25 (s, 1H, Ar-H), 4.25-4.22 (t, 6H, -OCH2), 4.10-4.08 (t, 6H, -CH2); HRMS (ESI + ): [M+H] + The calculation yields 415.06, and the result is 415.07.

[0023] (3) Intermediate 2 was added to N,N-dimethylformamide (DMF), followed by intermediate 1 and K2CO3. The ratio of intermediate 2, intermediate 1, K2CO3 and DMF was 10 mmol: 55 mmol: 23 mmol: 90 mL. The mixture was stirred at 55 °C for 28 h. The reaction solution was cooled to room temperature, diluted with water, and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined, washed with water, dried, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 50) to obtain the resveratrol derivative (yield 78.5%). 1 H NMR (C 59 H 45 N3O9, 400 MHz, d6-DMSO) δ 8.75-8.73 (d, 3H, Ar-H), 8.15-8.13(d, 3H, Ar-H), 7.82-7.79 (d, 2H, Ar-H), 7.67-7.65 (d, 6H, Ar-H), 7.52-7.50(t, 3H, Ar-H), 7.28 (s, 3H, -CH), 7.10-7.06 (m, 5H, Ar-H), 6.92-6.89 (d, 1H,CH), 6.86-6.83 (m, 5H, Ar-H), 6.83-6.82 (d, 1H, CH), 6.25 (s, 1H, Ar-H),4.45-4.42 (t, 6H, -OCH2), 3.90-3.87 (t, 6H, -CH2); HRMS (ESI + ): [M+H] + The calculation yields 940.32, and the result is 940.34.

[0024] The present application also provides a preparation method of the rutin composition for treating arthritis, and the specific steps are as follows: mixing the raw materials according to the weight parts to obtain the rutin composition.

[0025] Example 2 A rutin composition for treating arthritis, which comprises the following active ingredients in parts by weight: rutin 20 parts, typhaneoside 10 parts, and aescymenol derivative 8 parts.

[0026] The aescymenol derivative is prepared by the following process: (1) 2-indolone, furfural and a catalyst of piperidine are added into ethanol, and the amount ratio of the 2-indolone, furfural, catalyst of piperidine and ethanol is 10 mmol: 11 mmol: 0.1 mmol: 15 mL, and reflux reaction is carried out for 4 h; after the reaction is completed, the product is collected by filtration and washed with ethanol, and then purified by silica gel column chromatography (chloroform / methanol = 9:1) to obtain intermediate 1 (yield 85.0%); 1 H NMR and HRMS (ESI + ) results are the same as those of Example 1.

[0027] (2) aescymenol is added into water, and then sodium hydroxide, 1,2-dichloroethane and tetrabutylammonium bromide are added, and the amount ratio of the aescymenol, sodium hydroxide, 1,2-dichloroethane, tetrabutylammonium bromide and water is 20 mmol: 75 mmol: 180 mmol: 2 mmol: 15 mL; stirring is carried out at 75°C for 14 h; after the reaction solution is cooled to room temperature, water is added for dilution, and then extraction is carried out with ethyl acetate (3×30 mL), the organic phases are combined, and then washed with water, dried, filtered, concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 95 / 5) to obtain intermediate 2 (yield 79.8%); 1 H NMR and HRMS (ESI + ) results are the same as those of Example 1.

[0028] (3) intermediate 2 is added into DMF, and then intermediate 1 and K2CO3 are added, and the amount ratio of the intermediate 2, intermediate 1, K2CO3 and DMF is 10 mmol: 50 mmol: 20 mmol: 80 mL; stirring is carried out at 50°C for 30 h; after the reaction solution is cooled to room temperature, water is added for dilution, and then extraction is carried out with ethyl acetate (3×30 mL), the organic phases are combined, and then washed with water, dried, filtered, concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 50) to obtain the aescymenol derivative (yield 77.5%); 1 H NMR and HRMS (ESI + ) results are the same as those of Example 1.

[0029] The present application also provides a preparation method of the rutin composition for treating arthritis, and the specific steps are as follows: mixing the raw materials according to the weight parts to obtain the rutin composition.

[0030] Example 3 A rutin composition for treating arthritis, which comprises the following active ingredients in parts by weight: rutin 25 parts, typhaneoside 18 parts, and aesculetin derivative 15 parts.

[0031] The aesculetin derivative is prepared by the following process: (1) 2-indolone, furfural and a catalyst of piperidine are added into ethanol, and the amount ratio of the 2-indolone, furfural, catalyst of piperidine and ethanol is 10 mmol: 12 mmol: 0.5 mmol: 20 mL, and reflux reaction is carried out for 5 hours; after the reaction is completed, the product is collected by filtration and washed with ethanol, and then purified by silica gel column chromatography (chloroform / methanol = 9:1) to obtain the intermediate 1 (the yield is 85.8%); 1 H NMR and HRMS (ESI + ) results are the same as those of Example 1.

[0032] (2) aesculetin is added into water, and then sodium hydroxide, 1,2-dichloroethane and tetrabutylammonium bromide are added, and the amount ratio of the aesculetin, sodium hydroxide, 1,2-dichloroethane, tetrabutylammonium bromide and water is 20 mmol: 80 mmol: 200 mmol: 2.2 mmol: 20 mL; stirring is carried out at 85°C for 12 hours; after the reaction liquid is cooled to room temperature, it is diluted with water and extracted with ethyl acetate (3×30 mL), and then the organic phase is combined, washed with water, dried, filtered, concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 95 / 5) to obtain the intermediate 2 (the yield is 80.2%); 1 H NMR and HRMS (ESI + ) results are the same as those of Example 1.

[0033] (3) the intermediate 2 is added into DMF, and then the intermediate 1 and K2CO3 are added, and the amount ratio of the intermediate 2, intermediate 1, K2CO3 and DMF is 10 mmol: 60 mmol: 25 mmol: 100 mL; stirring is carried out at 60°C for 24 hours; after the reaction liquid is cooled to room temperature, it is diluted with water and extracted with ethyl acetate (3×30 mL), and then the organic phase is combined, washed with water, dried, filtered, concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 50) to obtain the aesculetin derivative (the yield is 79.3%); 1 H NMR and HRMS (ESI + ) results are the same as those of Example 1.

[0034] The present application also provides a preparation method of the rutin composition for treating arthritis, and the specific steps are as follows: mixing raw materials according to weight parts to obtain the rutin composition.

[0035] Comparative Example 1 The present comparative example 1 is basically the same as example 1, except that the aphyllantherol derivative is replaced by aphyllantherol, and the rest is the same as example 1.

[0036] Comparative Example 2 The present comparative example 2 is basically the same as example 1, except that the typhaneoside is omitted, and the rest is the same as example 1.

[0037] Comparative Example 3 The present comparative example 3 is basically the same as example 1, except that the typhaneoside and the aphyllantherol derivative are omitted, and the rest is the same as example 1.

[0038] Test Example 1 Experimental Animals DBA / 1 mice were selected as experimental animals, male, body weight 18-22 g, a total of 90, adaptively fed in a SPF laboratory for 1 week, and the environmental temperature was controlled at 23-25 ℃ and the relative humidity was 55%.

[0039] 2. Animal grouping The above DBA / 1 mice were randomly divided into a blank control group, a model group, a positive control group, an example 1 group, an example 2 group, an example 3 group, a comparative example 1 group, a comparative example 2 group, and a comparative example 3 group, 10 in each group.

[0040] 3. Construction of collagen-induced rheumatoid arthritis (CIA) mouse model The bovine type II collagen (CII) was dissolved in 0.01M glacial acetic acid to prepare 4mg / mL bovine type II collagen, which was then fully emulsified with an equal volume of Freund's complete adjuvant as an antigen. After the mice were anesthetized by intraperitoneal injection of Cetirizine (50mg / kg), the fur on the tail root was removed, and 100µL of the antigen was injected into the tail root after disinfection for sensitization. On the 21st day, the same dose of antigen (4mg / mL bovine type II collagen fully emulsified with an equal volume of Freund's incomplete adjuvant) was injected for immune attack. The blank control group was injected with normal saline.

[0041] 4. Administration method From the 21st day, the example 1 group, the example 2 group, the example 3 group, the comparative example 1 group, the comparative example 2 group, and the comparative example 3 group were respectively given 30mg / kg of the corresponding rutin composition by gavage, and the blank control group and the model group were given an equal amount of normal saline; the positive control group was given 1mg / kg of methotrexate by gavage once a day for 28 consecutive days.

[0042] 5. Index detection (1) Mouse arthritis score: On the 14th, 21st and 28th day after administration, the severity of arthritis of mice in each group was observed and scored, and the scoring criteria are shown in Table 1; each mouse was scored with the total score of 4 feet, and each mouse had a maximum score of 16, and the higher the arthritis index score, the more severe the arthritis symptoms of the mouse. The scoring results are shown in Table 2.

[0043] Table 1 Mouse arthritis scoring criteria Table 2 Mouse arthritis index score after administration As can be seen from Table 2, compared with the blank control group, the arthritis index score of the model group mice increased significantly, joint lesions appeared, and the model was successfully constructed. After 28 days of treatment, the arthritis index scores of Examples 1-3 decreased significantly compared with the model group; and the treatment effect was similar to that of the positive control group. Compared with Example 1, the treatment effect of Comparative Example 1 was reduced to some extent by replacing the arodistomol derivative with arodistomol, and the treatment effect of Comparative Example 2 was reduced to some extent by omitting the typhaneoside; the treatment effect of Comparative Example 3 was reduced to some extent by omitting the typhaneoside and arodistomol derivative. Among them, the effect of Comparative Example 3 was the worst. It is proved that typhaneoside and arodistomol derivative can synergize with rutin to enhance the treatment effect of arthritis inflammation.

[0044] (2) Mouse hind foot thickness: At the same time as scoring the mouse arthritis, the thickness of the mouse bilateral hind feet was measured by vernier caliper, and the final results are recorded in Table 3.

[0045] Table 3 Mouse foot thickness (mm) As can be seen from Table 3, compared with the blank control group, the foot thickness of the model group mice increased significantly, indicating that the swelling degree of the mouse feet was significantly increased. After 28 days of treatment, the foot thickness of mice in Examples 1-3 decreased significantly and was lower than that of the model group; and the treatment effect was similar to that of the positive control group. Compared with Example 1, the effects of Comparative Examples 1-3 were poor. The above results show that typhaneoside and arodistomol derivative can synergize with rutin to improve the foot inflammation level of CIA model mice and enhance the treatment effect of arthritis inflammation.

[0046] (3) Enzyme-linked immunosorbent assay (ELISA) to detect the level of anti-CII antibody in mouse serum: 3 hours after the last administration, the mice were anesthetized by intraperitoneal injection of su-tai (50 mg / kg), the eyeball was removed to take blood, centrifuged at 10,000 rpm for 10 min, the supernatant was taken, and the total immunoglobulin G (IgG) ELISA kit was operated according to the instructions, and the absorbance value of the sample at 450 nm / 650 nm was measured. The experimental results are shown in Table 4.

[0047] Table 4 From Table 4, it can be seen that, compared with the blank control group, the absorbance value of total immunoglobulin G of the model group mice was significantly increased. Compared with the model group, after treatment, the total immunoglobulin G levels of the mice in Examples 1-3 groups were significantly reduced, and were lower than those of the model group, indicating that the rutin composition obtained by the application could reduce the collagen-specific antibody level in the serum of the arthritis model mice. Moreover, compared with Example 1, the effects of Comparative Examples 1-3 were relatively poor. The above results show that the rutin composition of Examples 1-3 of the application can antagonize the increase of the serum antibody level of the arthritis model mice.

[0048] Finally, it should be pointed out that the above examples are only used to illustrate the technical solutions of the application, and not to limit it. The basic principles and main features of the application have been described above with specific embodiments, and some modifications or replacements can be made on the basis of the application, but these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the application claimed.

Claims

1. A rutin composition for the treatment of arthritis, characterized in that, The rutin composition comprises the following active ingredients by weight: rutin 20-25 parts, typhaneoside 10-18 parts, and orobanchyl alcohol derivative 8-15 parts; the orobanchyl alcohol derivative has the following structural formula: 。 2. The rutin composition for treating arthritis according to claim 1, wherein The orobanchyl alcohol derivative is prepared by the following process: (1) 2-indolone and furfural are added into a solvent, and a catalyst is added, and reflux reaction is carried out; after the reaction is completed, filtration, washing, purification are carried out to obtain intermediate 1; (2) orobanchyl alcohol is added into water, and sodium hydroxide, 1,2-dichloroethane and tetrabutylammonium bromide are added, and stirring reaction is carried out; after the reaction solution is extracted, the organic phases are combined, and the organic phase is washed with water, dried, filtered, concentrated, and purified to obtain intermediate 2; (3) intermediate 2 is added into a solvent, and intermediate 1 and K2CO3 are added, and stirring reaction is carried out; after the reaction solution is extracted, the organic phases are combined, and the organic phase is washed with water, dried, filtered, concentrated, and purified to obtain the orobanchyl alcohol derivative.

3. The rutin composition for treating arthritis according to claim 2, characterized in that, In step (1), the amount ratio of 2-indolone, furfural, catalyst, and solvent is 10 mmol: 11-12 mmol: 0.1-0.5 mmol: 15-20 mL, the catalyst is piperidine, and the solvent is ethanol.

4. The rutin composition for treating arthritis according to claim 2, wherein In step (1), the reflux reaction time is 4-5 h.

5. The rutin composition for treating arthritis according to claim 2, wherein In step (2), the amount ratio of orobanchyl alcohol, sodium hydroxide, 1,2-dichloroethane, tetrabutylammonium bromide, and water is 20 mmol: 75-80 mmol: 180-200 mmol: 2-2.2 mmol: 15-20 mL.

6. The rutin composition for treating arthritis according to claim 2, wherein In step (2), the reaction temperature is 75-85°C, and the reaction time is 12-14 h.

7. The rutin composition for treating arthritis according to claim 2, wherein In step (3), the amount ratio of intermediate 2, intermediate 1, K2CO3, and solvent is 10 mmol: 50-60 mmol: 20-25 mmol: 80-100 mL; the solvent is DMF.

8. The rutin composition for treating arthritis according to claim 2, wherein In step (3), the stirring reaction temperature is 50-60°C, and the stirring reaction time is 24-30 h.

9. A process for the preparation of a rutin composition for the treatment of arthritis according to any one of claims 1-8, characterized in that, The following steps are included: the raw materials are mixed according to weight parts to obtain the product.

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