Patent Application
20220143125
The present invention relates to the technical field of traditional Chinese medicine, and in particular, to Fuke Qianjin Capsules and a quality control method therefor.
Fuke Qianjin Capsule is a medicine made from 8 medicinal materials of Radix Et Caulis Flemingiae, Radix Rosa Laevigata, Herba Andrographis, Caulis Mahoniae, Zanthoxylum dissitum Hemsl., Radix Angelicae Sinensis, Caulis Spatholobi, and Radix Codonopsis. Its effect is to clear heat and eliminate dampness, and replenish and benefit qi and blood. It is used for leukorrheal diseases and abdominal pain caused by damp-heat stasis obstruction, symptoms of which include large quantity of leucorrhea, yellow in colour qualitative thick, and stinky, lower abdomen pain, lumbosacral soreness, and fatigued spirit and lack of strength; and chronic pelvic inflammatory disease, endometritis, and chronic cervicitis with the symptoms described above.
In the existing Pharmacopoeia of the People's Republic of China, for the content identification of the Fuke Qianjin Capsules, only total content of andrographolide and dehydroandrographolide is limited. Specifically, in each capsule, based on the total amount of andrographolide (C20H30O5) and dehydroandrographolide (C20H28O4), the content of Herba Andrographis shall not be less than 1.6 mg. Among compatible ingredients of the Fuke Qianjin Capsule, Herba Andrographis is only used as the minister drug, and obviously, the existing standards do not regulate other important ingredients in the Fuke Qianjin Capsule, so that it is difficult to control the consistency of effects between different batches of products of the Fuke Qianjin Capsules. Although relatively consistent products have been obtained through formulas and preparation methods, a more effective method is still needed to improve the therapeutic effect of the Fuke Qianjin Capsules.
The technical problem to be solved by the present invention is to overcome the above-mentioned shortcomings and deficiencies of the prior art, to provide Fuke Qianjin Capsules and a quality control method therefor. Compared with the existing Fuke Qianjin Capsules, the Fuke Qianjin Capsules prepared by the quality control method of the present invention are more stable in terms of effect consistency, and have a better clinical treatment effect than the existing Fuke Qianjin Capsules.
An objective of the present invention is to provide a quality control method for Fuke Qianjin Capsules.
Another objective of the present invention is to provide Fuke Qianjin Capsules.
The above-described objectives of the present invention are realized by the following technical solutions.
A quality control method for Fuke Qianjin Capsules, including the following steps:
using Radix Et Caulis Flemingiae, Radix Rosa Laevigata, Herba Andrographis, Radix Angelicae Sinensis, Caulis Mahoniae, Zanthoxylum dissitum Hemsl., Caulis Spatholobi and Radix Codonopsis as raw materials;
S1, first extracting the Radix Angelicae Sinensis by ethanol percolating to prepare a cream;
S2, extracting the Herba Andrographis by ethanol refluxing to prepare a cream;
S3, boiling and extracting the Caulis Mahoniae and the Zanthoxylum dissitum Hemsl. with water twice, after filtering, combining two filtrates to prepare a cream;
S4, boiling and extracting the Radix Et Caulis Flemingiae, the Radix Rosa Laevigata, the Caulis Spatholobi, and the Radix Codonopsis with water once and then filtering, boiling with water a filtered residue with a Radix Angelicae Sinensis residue produced in the step S1, an Herba Andrographis residue produced in the step S2 and a Caulis Mahoniae and Zanthoxylum dissitum Hemsl. residue produced in the step S3 together, filtering, combining filtrates to prepare a cream; and
S5, combining the four creams of the steps S1, S2, S3, S4, mixing, controlling a content of at least one of Z-ligustilide and Z-3-butylidenephthalide, and a total amount of andrographolide and dehydroandrographolide in the cream to reach a standard content, spray drying, and then pelletizing, canning to obtain the Fuke Qianjin Capsules. According to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the mass of each Fuke Qianjin Capsule is 0.4 mg.
In the Fuke Qianjin Capsules, the Radix Et Caulis Flemingiae and the Caulis Mahoniae clear heat and remove toxicity, eliminate dampness and arrest leucorrhoea, and together serve as a sovereign drug. The Herba Andrographis and the Zanthoxylum dissitum Hemsl. clear heat and remove toxicity, cool the blood and relieve swelling, eliminate dampness and arrest leucorrhoea, and serve as a minister drug. The Caulis Spatholobi and the Radix Angelicae Sinensis nourish the blood and promote blood circulation. The Radix Codonopsis benefits qi and strengthens spleen, and promotes transportation and digestion of water-dampness to arrest leucorrhoea. The Radix Rosa Laevigata arrests spontaneous emission and leucorrhoea, and serves as an assistant drug.
Traditional Chinese medicine compound is a hierarchical and structured organic whole. Its effect is not a simple addition of individual medicines, but a result of the mutual cooperation of multiple active ingredients. In the existing Pharmacopoeia, the standard regulations for the Fuke Qianjin Capsules only limit that andrographolide and dehydroandrographolide are from the minister drug (from Herba Andrographis), but do not limit types and contents of other active ingredients. In the actual production process, in quality detection and control, only andrographolide and dehydroandrographolide were detected, but types and contents of other active ingredients were not detected. As we all know, a content of an active ingredient in an extract of a traditional Chinese medicinal material is affected by the planting area of the medicinal material and the extraction method, so even if the extract is prepared according to the same raw material formula, a content of a specific active ingredient is not the same. Therefore, the content of other active ingredients that is not detected between different batches of the Fuke Qianjin Capsules are not uniform, resulting in large differences in their efficacy.
In order to ensure that the prepared Fuke Qianjin Capsules have the same efficacy in the actual production process, the inventors have found after many experiments that in addition to andrographolide and dehydroandrographolide, by controlling the contents of the six active ingredients of genistin, jatrorrhizine, palmatine, berberine, Z-ligustilide and Z-3-butylidenephthalide in the product within a certain range, the effect of the produced product is better, the efficacy of different batches is consistent, and the clinical treatment effect is improved. Due to the various influencing factors in a preparation process of a traditional Chinese medicine, the content control of core ingredients in the traditional Chinese medicine can overcome the problem of fluctuations in ingredient content between different batches. As a result, a new quality control method for the Fuke Qianjin Capsules is proposed. Using the quality control method, it can ensure that the contents of the five active ingredients in different batches of products are relatively consistent, and the consistency of the product's efficacy is relatively stable.
Preferably, in the step S5, per milligram of the cream, the content of the Z-ligustilide is not less than 0.01 mg and/or the content of the Z-3-butylidenephthalide is not less than 0.00017 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 0.01 mg.
Preferably, in the step S5, per milligram of the cream, the content of the Z-ligustilide is not less than 0.015 mg and/or the content of the Z-3-butylidenephthalide is not less than 0.00025 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 0.015 mg.
Preferably, in the step S5, per milligram of the cream, the content of the Z-ligustilide is 0.0165 mg to 0.022 mg and/or the content of the Z-3-butylidenephthalide is 0.00036 mg to 0.0006 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 0.02 mg.
A quality control method for Fuke Qianjin Capsules, including the following steps:
using Radix Et Caulis Flemingiae, Radix Rosa Laevigata, Herba Andrographis, Radix Angelicae Sinensis, Caulis Mahoniae, Zanthoxylum dissitum Hemsl., Caulis Spatholobi and Radix Codonopsis as raw materials;
S1, first extracting the Radix Angelicae Sinensis by ethanol percolating to prepare a cream;
S2, extracting the Herba Andrographis by ethanol refluxing to prepare a cream;
S3, boiling and extracting the Caulis Mahoniae and the Zanthoxylum dissitum Hemsl. with water twice, after filtering, combining two filtrates to prepare a cream;
S4, boiling and extracting the Radix Et Caulis Flemingiae, the Radix Rosa Laevigata, Caulis Spatholobi, and the Radix Codonopsis with water once and then filtering, boiling with water a filtered residue with a Radix Angelicae Sinensis residue produced in the step S1, an Herba Andrographis residue produced in the step S2 and a Caulis Mahoniae and Zanthoxylum dissitum Hemsl. residue produced in the step S3 together, filtering, combining filtrates to prepare a cream; and
S5, combining the four creams of the steps S1, S2, S3, S4, mixing, controlling a content of Z-ligustilide, and/or a content of Z-3-butylidenephthalide, and/or a content of genistin, and/or a content of jatrorrhizine, and/or a content of palmatine, and/or a content of berberine, and a total amount of andrographolide and dehydroandrographolide in the cream to reach a standard content, spray drying, and then pelletizing, canning to obtain the Fuke Qianjin Capsules.
Preferably, in the step S5, per milligram of the cream, the content of the genistin is not less than 0.00015 mg, the content of the jatrorrhizine is not less than 0.0004 mg, the content of the palmatine is not less than 0.00038 mg, the content of the berberine is not less than 0.0004 mg, the content of the Z-ligustilide is not less than 0.01 mg, the content of the Z-3-butylidenephthalide is not less than 0.00017 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 0.01 mg.
Preferably, in the step S5, per milligram of the cream, the content of the genistin is not less than 0.0002 mg, the content of the jatrorrhizine is not less than 0.0006 mg, the content of the palmatine is not less than 0.0006 mg, the content of the berberine is not less than 0.0006 mg, the content of the Z-ligustilide is not less than 0.015 mg, the content of the Z-3-butylidenephthalide is not less than 0.00025 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 0.015 mg.
Preferably, in the step S5, it is controlled that per milligram of the cream, the content of the genistin is 0.0025 mg to 0.0035 mg, the content of the jatrorrhizine is 0.009 mg to 0.015 mg, the content of the palmatine is 0.007 mg to 0.01 mg, the content of the berberine is 0.0085 mg to 0.01 mg the content of the Z-ligustilide is 0.0165 mg to 0.022 mg, the content of the Z-3-butylidenephthalide is 0.00036 mg 0.0006 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 0.02 mg.
Preferably, in each of the Fuke Qianjin Capsules prepared by said method (according to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the mass of each Fuke Qianjin Capsule is 0.4 mg), the genistin is not less than 0.025 mg, the jatrorrhizine is not less than 0.08 mg, the palmatine is not less than 0.06 mg, the berberine is not less than 0.08 mg, the Z-ligustilide is not less than 2.0 mg, the Z-3-butylidenephthalide is not less than 0.03 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 1.6 mg.
Preferably, in each of the Fuke Qianjin Capsules prepared by said method (according to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the mass of each Fuke Qianjin Capsule is 0.4 mg), the genistin is not less than 0.025 mg, the jatrorrhizine is not less than 0.08 mg, the palmatine is not less than 0.060 mg, the berberine is not less than 0.08 mg, the Z-ligustilide is not less than 2.0 mg, the Z-3-butylidenephthalide is not less than 0.03 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 1.9 mg.
Preferably, in each of the Fuke Qianjin Capsules prepared by said method (according to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the mass of each Fuke Qianjin Capsule is 0.4 mg), the genistin is not less than 0.032 mg, the jatrorrhizine is not less than 0.12 mg, the palmatine is not less than 0.075 mg, the berberine is not less than 0.12 mg, the Z-ligustilide is not less than 2.6 mg, the Z-3-butylidenephthalide is not less than 0.04 mg, and the total amount of the andrographolide and the dehydroandrographolide is not less than 2.5 mg.
Preferably, the total amount of the andrographolide and the dehydroandrographolide in each of the Fuke Qianjin Capsules is not less than 3.5 mg.
Preferably, each of the Fuke Qianjin Capsules prepared by said method contains 0.04 mg to 0.06 mg of the genistin, 0.165 mg to 0.20 mg of the jatrorrhizine, 0.090 mg to 0.130 mg of the palmatine, 0.13 mg to 0.18 mg of the berberine, 3.0 mg to 3.65 mg of the Z-ligustilide, 0.065 mg to 0.105 mg of the Z-3-butylidenephthalide, and the total amount of the andrographolide and the dehydroandrographolide is not less than 3.5 mg.
Preferably, in the step S5, said controlling makes the contents of the genistin, the jatrorrhizine, the palmatine, the berberine, the Z-ligustilide and the Z-3-butylidenephthalide after the 4 creams in the steps S1 to S4 are mixed reach a required range by adjusting an extraction process of the steps S1, S2, S3, S4 or a source of the raw materials.
A detection method adopted in the step S5 is HPLC detection.
Preferably, a preparation process of described in the step S5 is: taking 1 g cream, adding 200 mL of 75% formalin to dissolve, and then taking 2 mL of a dissolving solution and diluting to 10 mL with 75% formalin, passing through a 0.45 μm microporous membrane, and taking the filtrate as a solution to be tested; using Kromasil 100-5-C18 chromatographic column (250 mm×4.6 mm, 5 μm), with mobile phases using acetonitrile as an A phase and 0.1% phosphoric acid aqueous solution as a B phase, gradient eluting, with a flow rate being 1.0 mL·min-1, a detection wavelength being 254 nm, a column temperature being (30±0.5°) C., and an injection volume being 10 μL.
Preferably, the method of the step S1 is that crushing the Radix Angelicae Sinensis into coarse powder, passing through an 8-mesh sieve, infiltrating with ethanol with a mass concentration of 70%, placing in a percolation cylinder, immersing for 48 hours, percolating at a rate of 0.2 mL/min, and collecting 10 times the amount of percolation liquid, concentrating to obtain the Radix Angelicae Sinensis cream with a relative density of 1.1 (80° C.).
Preferably, the method of the step S2 is that: crushing the Herba Andrographis through a 100-mesh sieve, adding ethanol with a mass concentration of 90% according to a material-to-liquid ratio of 1:9 to extract by heating and reflux, refluxing for 4 hours, separating an extracting solution, and then refluxing the residue according to a material-liquid ratio of 1:8 for 3.5 hours, separating an extract solution, combining the two extract solutions, filtering, and concentrating to obtain the Herba Andrographis cream with a relative density of 1.1 (80° C.).
Preferably, the method of the step S3 is that boiling and extracting the Caulis Mahoniae and the Zanthoxylum dissitum Hemsl. with water twice, adding water 12 times the total weight of the two-flavored Chinese medicine for the first time and decocting for 4 hours, releasing an decoction, and adding water 10 times the amount of water for the second time and decocting for 3 hours, releasing an decoction, combining the decoctions, filtering, and concentrating an filtrate to obtain a cream with a relative density of 1.1 (80° C.).
Preferably, the method of the step S4 is that boiling the Radix Et Caulis Flemingiae, the Radix Rosa Laevigata, the Caulis Spatholobi, and the Radix Codonopsis according to a material-to-liquid ratio of 1:10 for 3.5 hours, extracting once and filtering, and concentrating an filtrate to obtain a cream with a relative density of 1.1 (80° C.), combining the filtered residue with the Radix Angelicae Sinensis residue generated in step S2, the Herba Andrographis residue generated in step S2, and the residue generated in step S3, and adding 4 times the volume of water to decoct for 3 hours, filtering, and concentrating to obtain a cream with a relative density of 1.1 (80° C.).
Preferably, the genistin is from the raw medicinal material Radix Et Caulis Flemingiae of the Fuke Qianjin Capsules, the jatrorrhizine, the palmatine and the berberine are from the raw medicinal material Caulis Mahoniae of the Fuke Qianjin Capsules, the Z-ligustilide and the Z-3-butylidenephthalide are from the raw medicinal material Radix Angelicae Sinensis of the Fuke Qianjin Capsules, and the andrographolide and the dehydroandrographolide are from the raw medicinal material Herba Andrographis of the Fuke Qianjin Capsules.
Preferably, dosage of the Radix Angelicae Sinensis, the Radix Codonopsis, the Herba Andrographis and the Zanthoxylum dissitum Hemsl. is each 9% of a total amount of the medicinal materials of the Fuke Qianjin Capsules; and dosage of the Radix Rosa Laevigata, the Caulis Spatholobi, the Caulis Mahoniae and the Radix Et Caulis Flemingiae is each 16% of the total amount of the medicinal materials of the Fuke Qianjin Capsules.
The present invention also claims for protection for Fuke Qianjin Capsules prepared by the above quality control method and use thereof in preparation of drugs for treating a gynecological disease.
Preferably, the gynecological disease is chronic pelvic inflammatory disease, chronic adnexitis or endometritis.
The present invention also claims for protection for Fuke Qianjin Capsules, which are made of Radix Et Caulis Flemingiae, Caulis Mahoniae, Herba Andrographis, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Radix Angelicae Sinensis, Radix Codonopsis and Radix Rosa Laevigata as raw materials, each of the Fuke Qianjin Capsules contains not less than 2.0 mg of Z-ligustilide; preferably contains 2.6 mg of the Z-ligustilide; and more preferably contains 3.0 mg to 3.65 mg of the Z-ligustilide;
and a total amount of andrographolide and dehydroandrographolide is not less than 1.9 mg; preferably, the total amount of the andrographolide and the dehydroandrographolide is not less than 2.5 mg; and preferably, the total amount of the andrographolide and the dehydroandrographolide is not less than 3.5 mg.
Fuke Qianjin Capsules, made of Radix Et Caulis Flemingiae, Caulis Mahoniae, Herba Andrographis, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Radix Angelicae Sinensis, Radix Codonopsis and Radix Rosa Laevigata as raw materials, each of the Fuke Qianjin Capsules contains not less than 0.03 mg of Z-3-butylidenephthalide; preferably contains not less than 0.04 mg of the Z-3-butylidenephthalide; and preferably contains 0.065 mg to 0.105 mg of the Z-3-butylidenephthalide;
and a total amount of andrographolide and dehydroandrographolide is not less than 1.9 mg; preferably, the total amount of the andrographolide and the dehydroandrographolide is not less than 2.5 mg; and preferably, the total amount of the andrographolide and the dehydroandrographolide is not less than 3.5 mg.
Fuke Qianjin Capsules, made of Radix Et Caulis Flemingiae, Caulis Mahoniae, Herba Andrographis, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Radix Angelicae Sinensis, Radix Codonopsis and Radix Rosa Laevigata as raw materials, each of the Fuke Qianjin Capsules contains not less than 2.0 mg of Z-ligustilide, and not less than 0.03 mg of Z-3-butylidenephthalide; and a total amount of andrographolide and dehydroandrographolide is not less than 1.9 mg;
preferably each of the Fuke Qianjin Capsules contains not less than 2.6 mg of the Z-ligustilide, not less than 0.04 mg of the Z-3-butylidenephthalide, and the total amount of the andrographolide and the dehydroandrographolide is not less than 2.5 mg;
preferably each of the Fuke Qianjin Capsules contains 3.0 mg to 3.65 mg of the Z-ligustilide, and 0.065 mg to 0.105 mg of the Z-3-butylidenephthalide, and the total amount of the andrographolide and the dehydroandrographolide is not less than 3.5 mg.
Fuke Qianjin Capsules, made of Radix Et Caulis Flemingiae, Caulis Mahoniae, Herba Andrographis, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Radix Angelicae Sinensis, Radix Codonopsis and Radix Rosa Laevigata as raw materials, each of the Fuke Qianjin Capsules (according to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the mass of each Fuke Qianjin Capsule is 0.4 mg) contains not less than 0.025 mg of genistin, not less than 0.08 mg of jatrorrhizine, not less than 0.060 mg of palmatine, not less than 0.08 mg of berberine, not less than 2.0 mg of Z-ligustilide, and not less than 0.03 mg of Z-3-butylidenephthalide, and a total amount of the andrographolide and the dehydroandrographolide is not less than 1.9 mg.
Preferably, each of the Fuke Qianjin Capsules contains not less than 0.032 mg of the genistin, not less than 0.12 mg of the jatrorrhizine, not less than 0.075 mg of the palmatine, not less than 0.12 mg of the berberine, not less than 2.6 mg of the Z-ligustilide, and not less than 0.04 mg of the Z-3-butylidenephthalide, and the total amount of the andrographolide and the dehydroandrographolide is not less than 2.5 mg.
Preferably, in each of the Fuke Qianjin Capsules, the total amount of the andrographolide and the dehydroandrographolide is not less than 3.5 mg.
Preferably, each of the Fuke Qianjin Capsules contains 0.04 mg to 0.06 mg of the genistin, 0.165 mg to 0.20 mg of the jatrorrhizine, 0.090 mg to 0.130 mg of the palmatine, 0.13 mg to 0.18 mg of the berberine, 3.0 mg to 3.65 mg of the Z-ligustilide, and 0.065 mg to 0.105 mg of the Z-3-butylidenephthalide, and the total amount of the andrographolide and the dehydroandrographolide is not less than 3.5 mg.
Preferably, each of the Fuke Qianjin Capsules contains 0.045 mg to 0.056 mg of the genistin, 0.175 mg to 0.195 mg of the jatrorrhizine, 0.10 mg to 0.130 mg of the palmatine, 0.16 mg to 0.178 mg of the berberine, 3.30 mg to 3.65 mg of the Z-ligustilide, 0.080 mg to 0.105 mg of the Z-3-butylidenephthalide, 2.5 mg to 4.15 mg of the andrographolide and 1.0 mg to 1.3 mg of the dehydroandrographolide.
Additionally preferably, the contents of the genistin, the jatrorrhizine, the palmatine, the berberine, the Z-ligustilide, the Z-3-butylidenephthalide, and the total amount of the andrographolide and the dehydroandrographolide are determined by HPLC detection.
Preferably, a test sample for the HPLC detection is prepared by the following method: taking out 1.00 g of contents of Fuke Qianjin Capsules and placing in a triangular flask, accurately adding 20 mL of 75% formalin, and ultrasonically extracting for (30±5) minutes, after cooling to room temperature, using 75% formalin to make up a mass loss, passing through a 0.45 μm microporous membrane, and taking the filtrate as a solution to be tested.
Preferably, the HPLC detection is performed according to the following conditions: using Kromasil 100-5-C18 chromatographic column (250 mm×4.6 mm, 5 μm), with mobile phases using acetonitrile as an A phase and 0.1% phosphoric acid aqueous solution as a B phase, gradient eluting, with a flow rate being 1.0 mL·min-1, a detection wavelength being 254 nm, a column temperature being (30±0.5°) C., and an injection volume being 10 μL.
Preferably, the genistin is from the raw medicinal material Radix Et Caulis Flemingiae of the Fuke Qianjin Capsules, the jatrorrhizine, the palmatine and the berberine are from the raw medicinal material Caulis Mahoniae of the Fuke Qianjin Capsules, the Z-ligustilide and the Z-3-butylidenephthalide are from the raw medicinal material Radix Angelicae Sinensis of the Fuke Qianjin Capsules, and the andrographolide an the dehydroandrographolide are from the raw medicinal material Herba Andrographis of the Fuke Qianjin Capsules.
Preferably, dosage of the Radix Angelicae Sinensis, the Radix Codonopsis, the Herba Andrographis and the Zanthoxylum dissitum Hemsl. is each 9% of a total amount of the medicinal materials of the Fuke Qianjin Capsules; and dosage of the Radix Rosa Laevigata, the Caulis Spatholobi, the Caulis Mahoniae and the Radix Et Caulis Flemingiae is each 16% of the total amount of the medicinal materials of the Fuke Qianjin Capsules.
The present invention also claims for protection for Fuke Qianjin Capsules prepared by the above quality control method and use thereof in preparation of drugs for treating a gynecological disease. Preferably, the gynecological disease is chronic pelvic inflammatory disease, chronic adnexitis or endometritis.
Compared with the prior art, the present invention has the following beneficial effects.
The present invention has established a new standard for controlling quality of the Fuke Qianjin Capsules through an analysis of chemical ingredients in the Fuke Qianjin Capsules. This standard adds a variety of core ingredient content to the existing pharmacopoeia standards. According to the Fuke Qianjin Capsules made in this range, the consistency of effects between different batches is more stable. Moreover, the more the types of core ingredients are limited, the more stable the consistency of the drug effect. Compared with the prior art, the Fuke Qianjin Capsules provided by the present invention has a better clinical treatment effect.
Further, in addition to controlling the contents of andrographolide and dehydroandrographolide in the product, a detection process is added to the preparation process of the product to control the contents of genistin, jatrorrhizine, palmatine, beberine, Z-ligustilide and Z-3-butylidenephthalide in the product to ensure that the contents of these 6 active ingredients in the product is within a certain range, thereby ensuring that the consistency of the efficacy of the product between different batches is more stable.
At the same time, the present invention adopts HPLC method for detection, which not only has high accuracy of detection results, but also rapid and simple detection process, which is convenient for the actual production process of the Fuke Qianjin Capsules of the present invention to simultaneously detect and monitor contents of multiple active ingredients in the original preparation process, which is conducive to the implementation of the new standard.
The present invention is further described in detailed below in combination with specific embodiments, which are only used to explain the present invention, and are not used to limit the scope of the present invention. Unless otherwise specified, test methods used in the following embodiments are conventional methods. The materials and reagents used, unless otherwise specified, are commercially available reagents and materials.
Formula of Fuke Qianjin Capsules: dosage of Radix Angelicae Sinensis, Radix Codonopsis, Herba Andrographis and Zanthoxylum dissitum Hemsl. was each 9% of a total amount of the medicinal materials; and dosage of Radix Rosa Laevigata, Caulis Spatholobi, Caulis Mahoniae and Radix Et Caulis Flemingiae was each 16% of the total amount of the medicinal materials. The total amount of the medicinal materials was 500 kg. The product was prepared by the following methods:
(1) the Radix Angelicae Sinensis was first extracted by ethanol percolating to prepare a cream: the Radix Angelicae Sinensis was crushed into coarse powder, passed through an 8-mesh sieve, infiltrated with ethanol with a mass concentration of 70%, placed in a percolation cylinder, immersed for 48 hours, percolated at a rate of 0.2 mL/min, and collected 10 times the amount of percolation liquid, concentrated to obtain a relative density of 1.1 (80° C.), which was the Radix Angelicae Sinensis cream;
(2) the Herba Andrographis was extracted by ethanol refluxing to prepare a cream: the Herba Andrographis was crushed through a 100-mesh sieve, ethanol with a mass concentration of 90% was added according to a material-to-liquid ratio of 1:9 to extract by heating and reflux, refluxed for 4 hours, an extracting solution was separated, and then the residue was refluxed according to a material-liquid ratio of 1:8 for 3.5 hours, an extract solution was separated, the two extract solutions was combined, filtered, and concentrated to obtain a relative density of 1.1 (80° C.), which was the Herba Andrographis cream;
(3) the Caulis Mahoniae and the Zanthoxylum dissitum Hemsl. were boiling extracted with water twice, water 12 times the total weight of the two-flavored Chinese medicine was added for the first time and decocted for 4 hours, an decoction was released; and water 10 times the total weight of the two-flavored Chinese medicine was added for the second time and decocted for 3 hours, an decoction was released, the decoctions were combined, filtered, and an filtrate was concentrated to a relative density of 1.1 (80° C.) to obtain a cream of the two medicines;
(4) the Radix Et Caulis Flemingiae, the Radix Rosa Laevigata, the Caulis Spatholobi, and the Radix Codonopsis were boiled according to a material-to-liquid ratio of 1:10 for 3.5 hours, extracted once and filtered, and an filtrate was concentrated to obtain a cream with a relative density of 1.1 (80° C.); then the filtered residue with the Radix Angelicae Sinensis residue generated in step S2, the Herba Andrographis residue generated in step S2, and the residue generated in step S3 were combined, and 4 times the volume of water was added to decoct for 3 hours, filtered, and an filtrate was concentrated to obtain a cream with a relative density of 1.1 (80° C.); and
(5) the four creams of the (1), (2), (3), (4) were combined and mixed for HPLC detection, per milligram of cream, a content of Z-ligustilide was controlled not less than 0.01 mg, and/or a content of Z-3-butylidenephthalide was controlled not less than 0.00017 mg, and/or a content of genistin was controlled not less than 0.00015 mg, and/or a content of jatrorrhizine was controlled not less than 0.0004 mg, and/or a content of palmatine was controlled not less than 0.0038 mg, and/or a content of berberine was controlled not less than 0.0004 mg, and a total amount of andrographolide and dehydroandrographolide was controlled not less than 0.01 mg, spray drying, pelletizing, and canning were carried out to obtain the Fuke Qianjin Capsules, and according to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the content of each Fuke Qianjin Capsule was 0.4 g.
When in the detection result of the mixed cream in the step (5), the content of any one of the 6 active ingredients (Z-ligustilide, Z-3-butylidenephthalide, genistin, jatrorrhizine, palmatine, berberine) failed to meet the above conditions, a part of sample was reserved for use.
When in the detection result of the mixed cream in the step (5), the contents of 6 active ingredients (Z-ligustilide, Z-3-butylidenephthalide, genistin, jatrorrhizine, palmatine, berberine) all failed to meet the above conditions, it was a comparative sample and was reserved for use.
The cause of the cream with fluctuating active ingredient content was analyzed, which may be caused by the difference in content of the active ingredients of the raw materials between different batches.
The Fuke Qianjin Capsules were prepared according to the above method, multiple batches of products were randomly selected from a long-term, large number of products, and HPLC detection was performed on each batch of the Fuke Qianjin Capsule products.
Preparation of a solution to be tested: Fuke Qianjin Capsules were randomly selected, 1.0 g of contents was taken out and placed in a triangular flask with a ground stopper, 20 mL of 75% formalin was accurately added, weighing was performed, ultrasonic extraction was performed for 30 minutes, after cooled to room temperature, 75% formalin was used to make up a mass loss, after passing through a 0.45 μm microporous membrane, a filtrate was taken as the solution to be tested.
Kromasil 100-5-C18 chromatographic column (250 mm×4.6 mm, 5 μm) was used, mobile phases were acetonitrile (A phase) and 0.1% phosphoric acid aqueous solution (B phase), gradient elution was performed, with a flow rate of 1.0 mL·min-1, a detection wavelength of 254 nm, a column temperature of 30° C., and an injection volume of 10 μL.
Fuke Qianjin Capsules, the steps (1) to (4) in the preparation process thereof are the same as in Embodiment 1, and the difference from Embodiment 1 is that in the step (5), per milligram of the cream, the content of the Z-ligustilide was controlled not less than 0.015 mg, and/or the content of the Z-3-butylidenephthalide was controlled not less than 0.00025 mg, and/or the content of the genistin was controlled not less than 0.002 mg, and/or the content of the jatrorrhizine was controlled not less than 0.006 mg, and/or the content of the palmatine was controlled not less than 0.006 mg, and/or the content of the berberine was controlled not less than 0.006 mg, and the total amount of the andrographolide and the dehydroandrographolide was controlled not less than 0.015 mg; spray drying, pelletizing and canning were carried out to obtain the Fuke Qianjin Capsules, and according to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the content of each Fuke Qianjin Capsule was 0.4 g.
The Fuke Qianjin Capsules were prepared according to the above method, multiple batches of products were randomly selected from a long-term, large number of products, and detection was performed on each batch of the Fuke Qianjin Capsules according to the HPLC detection method in Embodiment 1.
The HPLC detection results of the 6 batches randomly selected from the multiple batches of samples detected in Embodiment 1 and the 6 batches randomly selected in the Embodiment 2 are shown in Table 1.
The cream in which the contents of Z-ligustilide, Z-3-butylidenephthalide, genistin, jatrorrhizine, palmatine and berberine in Embodiment 1 were not within the range required by Embodiment 1 was selected and directly performed with spray drying, and then pelletizing, canning to obtain the Fuke Qianjin Capsules. According to the 2015 edition of the Pharmacopoeia of the People's Republic of China, the content of each Fuke Qianjin Capsule was 0.4 g. The same method was used for HPLC detection for each batch of the Fuke Qianjin Capsules.
The detection conditions and methods are the same as Embodiment 1, and the detection results are shown in Table 2.
It can be seen from Table 2 that in the Fuke Qianjin Capsules prepared according to the existing method, except that the content of andrographolide and dehydroandrographolide meets the requirements of the Pharmacopoeia, the contents of other active ingredients vary greatly between batches, which can easily lead to instability of drug efficacy.
Medicines or materials used: croton oil, provided by Nanjing Institute of Dermatology; carrageenan, produced by Wako Pure Chemical Industries, Ltd.; nutrient broth medium, product of Guangdong Huankai Microbial Technology Co., Ltd.; and mould medium, provided by China National Institute for the Control of Pharmaceutical and Biological Products.
The Fuke Qianjin Capsules prepared by Embodiment 1 (batch 1 and batch 6) and Comparative Example 1 (batch 1) were used as samples. Kunming mice of clean grade and SD rats used were provided by the Hunan Institute for Drug Control; Escherichia coli ATCC25922, Staphylococcus aureus ATCC25923, beta hemolytic streptococcus ATCC32172 were all provided by the Provincial Health and Epidemic Prevention Station, Candida albicans, isolated from the clinic, was provided by the Bacteria Room of the Clinical Laboratory Department of the Third Affiliated Hospital of Hunan Medical University.
1) In Vitro Antibacterial Test on Standard Bacteria
The results are shown in Table 3 to Table 5.
Escherichia
coli
Staphylococcus
aureus
streptococcus
Candida
albicans
Escherichia
coli
Staphylococcus
aureus
streptococcus
Candida
albicans
Escherichia
coli
Staphylococcus
aureus
streptococcus
Candida
albicans
== indicates that there is no bacterial growth in two repeated experiments, and +, ++, +++ indicate the degree of cell growth, respectively.
The experimental results of Table 3 to Table 5 show that: the Fuke Qianjin Capsules of Embodiment 1 and Comparative Example 1 have minimum inhibitory concentrations of 12.5, 6.25, 12.5, 12.5 for Escherichia coli, Staphylococcus aureus, beta hemolytic streptococcus and Candida albicans, respectively, and the inhibitory concentrations of the two are the same.
2) In Vitro Antibacterial Experiment on Clinically Isolated Pathogenic Bacteria
Escherichia coli, Staphylococcus aureus, beta hemolytic streptococcus, and Candida albicans were all isolated from clinical patients and identified by bacteriology, provided by the Bacteria Room of the Clinical Laboratory Department of the Third Affiliated Hospital of Hunan Medical University.
The results are shown in Table 6 to Table 8.
Escherichia coli
Staphylococcus aureus
streptococcus
Candida albicans
Escherichia coli
Staphylococcus aureus
streptococcus
Candida albicans
Escherichia coli
Staphylococcus aureus
streptococcus
Candida albicans
The experimental results of Table 6 to Table 8 show that the Fuke Qianjin Capsules prepared by Embodiment 1 and Comparative Example 1 have the same inhibitory effect on the four clinically isolated bacteria, and the effect is consistent.
3) Antibacterial Experiment In Vivo
Protective effect on Escherichia coli-infected mice: 150 Kunming mice, both male and female, weighing 20-22 g, were divided into 15 groups (n=10), five groups were used for the Fuke Qianjin Capsules of batch 1 of Embodiment 1, five groups were used for the Fuke Qianjin Capsules of batch 6 of Embodiment 1, and five groups were used for the Fuke Qianjin Capsules of batch 1 of Comparative Example 1, and the doses were 17.3, 24.7, 35.3, 50.4 and 72.0 g crude drug/kg.
Volume of intragastric administration was 0.5 mL/20 g, and each mouse in each group was injected with 0.5 mL of Escherichia coli (106 FGu/mL) decoction culture solution one hour after the administration. At 12 hours and 24 hours after the mice were infected with the bacteria, the mice were administered twice, and then observed for seven days, and the number of animal deaths was recorded. The peak of animal deaths was between 24 hours and 48 hours.
The experimental results in Table 9 show that the Fuke Qianjin Capsules prepared by Embodiment 1 and Comparative Example 1 have a certain protective effect on Escherichia coli-infected mice in the high-dose group.
4) Anti-Inflammatory Effect (Influence on Croton Oil-Induced Swelling in Mice)
Preparation of drugs to be used: the ready-to-use contents of Fuke Qianjin Capsules were taken and prepared into suspensions of different concentrations with distilled water for mice. Intragastric administration was performed once a day, and the liquid for intragastric administration was used it right after it was prepared.
100 mice, male, weighing 24-26 g, were equally divided into 10 groups (n=10), and were given different concentrations of liquid medicine by intragastric administration, and the control group was given with equal volume of distilled water by intragastric administration once a day for 7 days, 1 hour after the last administration was performed, the right ear of each mouse was applied with 0.1 mL of 2% croton oil (2% croton oil, 20% anhydrous ethanol, 5% distilled water and 73% diethyl ether), and no treatment was applied on the left ear of all mice. On the 7th day, 4 hours after the last administration, the mice were sacrificed by cervical dislocation, the ears were cut off, punched and weighed, and the swelling degree of each group of mice was calculated.
21 ± 1.1
23 ± 2.3
The results in Table 10 show that: compared with the control group, the Fuke Qianjin Capsules prepared by Embodiment 1 and Comparative Example 1 can both significantly inhibit croton oil-induced ear swelling in mice; however, the inhibition effect of the two batches in Embodiment 1 is significantly better than the Fuke Qianjin Capsules of Comparative Example 1.
5) Influence on Carrageenan-Induced Footpad Swelling in Rats
70 SD rats, males, were divided into seven groups (n=10), and they were given different concentrations of liquid medicine by intragastric administration, and the control group was given with the same volume of distilled water by intragastric administration, once a day for seven days, 1 hour after the last administration was performed, 0.1 mL of carrageenan was injected into the bottom of the right pedal of each rat to cause inflammation. 2 hours after the inflammation, it is administered again. In addition to measuring the size of the normal pedal before the inflammation, the size of pedal was measured every 1 hour after the inflammation for a total of 6 times, and the swelling degree was calculated.
The results in Table 11 show that: compared with the control group, the Fuke Qianjin Capsules prepared by Embodiment 1 and Comparative Example 1 can significantly inhibit the carrageenan-induced footpad swelling in rats; however, the two batches of the Fuke Qianjin Capsules in Embodiment 1 have better inhibitory effects on the carrageenan-induced footpad swelling in rats than the Fuke Qianjin Capsules of Comparative Example 1, especially in the 3-4 hours period after the administration.
6) Influence on Induced Painful Mice Induced by Acetic Acid
100 mice, half male and half male, weighing 20-22 g, were randomly divided into 10 groups (n=10), and they were given different concentrations of liquid medicine by intragastric administration, and the control group was given with the same volume of distilled water by intragastric administration, one hour after the administration, each mouse was injected with 0.2 mL of 0.6% acetic acid. 5 minutes after the injection, the recording was started, and the number of mouse writhing times in 10 minutes was recorded.
The results in Table 12 show that: compared with the control group, the Fuke Qianjin Capsules prepared by Embodiment 1 and Comparative Example 1 can significantly reduce the number of writhing times of painful mice induced by acetic acid; however, the effect of the two batches of the Fuke Qianjin Capsules in Embodiment 1 on the pain induced by acetic acid is significantly better than the Fuke Qianjin Capsules of Comparative Example 1.
7) Influence on Pain Threshold of Painful Mice Induced by Hot Plate Test
Pain threshold of a mouse was measured according to the following method: the mouse was placed on a 55±0.5° C. hot plate at a room temperature of 20±1° C., and the time from touching the hot plate to licking the rear foot was the pain threshold.
Then 100 female mice with a pain threshold of less than 30 seconds, weighing 20-22 g, were divided into 10 groups (n=10), and they were given by one-time intragastric administration according to the dose of painful mice induced by acetic acid, and according to the above measuring method, the pain thresholds were measured at 55±0.5° C. before administration and 30, 60 and 90 minutes after administration, and the results are shown in Table 13.
The results in Table 13 show that compared with the control group, the Fuke Qianjin Capsules of Embodiment 1 and Comparative Example 1 can significantly increase the pain threshold of painful mice induced by hot plate excitement; however, the percentage increase in pain threshold of painful mice induced by hot plate excitement of the two batches of Fuke Qianjin Capsules in Embodiment 1 is significantly higher than the Fuke Qianjin Capsules of Comparative Example 1.
8) Influence on Hemorrhagic Blood Deficiency Mice
110 mice, both male and female, weighing 20-22 g, were divided into 11 groups (n=10), First, blood was taken to measure normal values RBC (red blood cell) and HB (haemoglobin) of the mice, then, except for the normal control group, each mouse in the other groups was bled 0.5 mL from the orbital venous plexus, 24 hours later, blood was taken to measure the RBC and HB values of the mice, and then different doses of medicine were given by intragastric administration, once a day for seven days, 24 hours after the last administration, blood was taken from the orbital venous plexus of the mice to measure the RBC and HB values, and the results are shown in Table 14.
The results in Table 14 show that, compared with the control group, the fuke qianjin ta capsules of Embodiment 1 and Comparative Example 1 both have a significant blood-enriching effect on hemorrhagic blood deficiency mice; however, the two batches of Fuke Qianjin Capsules in Embodiment 1 have a significantly higher blood-enriching effect on hemorrhagic blood deficiency mice than the Fuke Qianjin Capsules of Comparative Example.
Throughout the above experimental results, it can be seen that the Fuke Qianjin Capsules of Embodiment 1 and Comparative Example 1 have the same in vitro minimal inhibitory concentration for Escherichia coli, Staphylococcus aureus, beta hemolytic streptococcus and Candida albicans, as well as the minimal inhibitory concentration of the above four clinically isolated bacteria.
However, in mice in vivo experiments, the Fuke Qianjin Capsules of the two batches in Embodiment 1 and in Comparative Example 1 can significantly inhibit croton oil-induced ear swelling in mice and carrageenan-induced footpad swelling in rats; reduce the number of writhing times in mice induced by acetic acid, and increase the pain threshold of painful mice induced by hot plate test; and can also have a significant blood-enriching effect on hemorrhagic blood deficiency mice; however, The above-mentioned effects of the Fuke Qianjin Capsules prepared in the two batches in Embodiment 1 have a certain degree of improvement compared with the Fuke Qianjin Capsules of Comparative Example 1, and the effects are better than the Fuke Qianjin Capsules of Comparative Example 1.
In order to compare whether there is a difference between the Fuke Qianjin Capsules that the present invention controls the content of multiple active ingredients and the Fuke Qianjin Capsules prepared by the original method (i.e., Comparative Example 1), a clinical trial was conducted. In accordance with the requirements of relevant new drug research, each test site has formulated clinical research principles, established diagnostic criteria, inclusion criteria, and rejection criteria, and used this as a guideline to collect observation cases. At the same time, method of taking medicine was followed in the instructions attached to the medicine during use, the medicine was started taking when seeing a doctor, 7 days as a course of treatment, two consecutive courses of treatment, and clinical symptoms and changes in symptoms were collected according to the planned clinical observation form.
Table 15 shows the clinical changes of 240 patients with chronic pelvic inflammatory disease in the Second Affiliated Hospital of Hunan University of Traditional Chinese Medicine, Hunan Academy of Traditional Chinese Medicine, and the First Affiliated Hospital of Hunan Medical University after taking qianjin capsules for two courses, divided into 3 groups, 80 patients in each group. The specific results are shown in Table 15.
The effect statistics of 240 patients with adnexitis taking Fuke Qianjin Capsules were listed in Tables 16 to 18, wherein Table 16 is the examination status of the 240 patients with adnexitis, Table 17 is the efficacy statistics, and Table 18 is the change in symptoms before and after taking the Fuke Qianjin Capsules.
It can be seen from Tables 15 to 18 that the Fuke Qianjin Capsules of the present invention is better than the Fuke Qianjin Capsules of Comparative Example 1 in the treatment of chronic pelvic inflammatory disease and adnexitis, wherein the effect of batch 6 of Embodiment is better than that of batch 1, and the effect of batch 1 of Embodiment is better than that of Comparative Example. It reflects that in addition to controlling the contents of Z-ligustilide, Z-3-butylidenephthalide, andrographolide and dehydroandrographolide, it also controls the contents of genistin, jatrorrhizine, palmatine and berberine within a standard range, and the therapeutic effect can be further improved.
According to the good results shown in the treatment of chronic pelvic inflammatory disease, we also compared the efficacy of the treatment of endometritis. Specifically, 608 patients with endometritis were selected as study subjects, aged between 30 and 40 years old, and the treatment plan was to give antibiotics combined with progesterone for treatment. 0.5 g of metronidazole was added into 250 mL of 0.9% sodium chloride solution, intravenously dripped, once every 8 hours, medroxyprogesterone was taken 4 mg/time, 2 times a day for 14 days after the end of menstruation on the 3rd day, and this is used as a blank control group. The observation group was given the Fuke Qianjin Capsules of Embodiment 1 (3 batches) and Embodiment 2 (3 batches) on the basis of the blank control group, and the control group was given the Fuke Qianjin Capsules prepared in Comparative Example 1 (3 batches) on the basis of the blank control group. The treatment results are shown in Tables 19 to 21.
Evaluation Criteria:
Markedly effective: the clinical symptoms disappeared, the menstruation returned to normal, and the ultrasound examination showed that the inflammation disappeared;
Effective: clinical symptoms got better, and the ultrasound examination showed that the inflammation got better and endometrium was thickened;
Noneffective: no improvement as described above.
From the data in Tables 19-21, it can be seen that compared to the original Fuke Qianjin Capsules that only control the active ingredient of Herba Andrographis, the Fuke Qianjin Capsules of the present invention that the contents of genistin, and/or jatrorrhizine, and/or palmatine, and/or berberine are controlled, the effectiveness of the Fuke Qianjin Capsules in the treatment of endometritis is improved. The specific performance is that the efficiency and the markedly effective rate have been improved, indicating that when the contents of genistin, and/or jatrorrhizine, and/or palmatine, and/or berberine are controlled, the Fuke Qianjin Capsules can interact better with antibiotics and progesterone.
From the consistency experiment of the above Embodiments and Comparative Example, it can be seen that in the production process, the detection of the ingredients of the mixed cream sample is added, and the control is within a reasonable range, so that the contents of the eight active ingredients in the obtained Fuke Qianjin Capsules can be controlled within a reasonable and narrow range, so that the consistency between batches of the prepared product is better, and the clinical treatment effect is improved.
Finally, it should be noted that the above Embodiments are only used to illustrate the technical solution of the present invention and not to limit the scope of protection of the present invention. For those of ordinary skill in the art, on the basis of the above description and ideas, other different forms of changes or variations can also be made, and it is not necessary and impossible to enumerate all the implementation here. All modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included in the scope of protection claimed in the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201811356104.0 | Nov 2018 | CN | national |
| 201811356105.5 | Nov 2018 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2020/072103 | 1/14/2020 | WO | 00 |
