CN111228330A - Stephanine-containing anti-inflammatory pharmaceutical composition and preparation method thereof - Google Patents
Stephanine-containing anti-inflammatory pharmaceutical composition and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the field of medicines, and particularly relates to an anti-inflammatory pharmaceutical composition containing stephanine and a preparation method thereof. The anti-inflammatory pharmaceutical composition provided by the invention comprises the following components in percentage by weight: 1-2% of rabdosia rubescens extract, 1-2% of andrographis paniculata extract, 2-3% of cepharanthine, 0.2-0.4% of essential oil, 0.1-0.3% of emulsifier and 92.3-95.7% of purified water. According to the anti-inflammatory pharmaceutical composition provided by the invention, the isoquinoline alkaloid compound-stephanine extracted from Stephania delavayi Diels is added, so that the anti-inflammatory performance of the pharmaceutical composition can be improved, no stimulation reaction is caused to a human body, and the possibility is provided for developing a new anti-inflammatory drug.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to an anti-inflammatory pharmaceutical composition containing stephanine and a preparation method thereof.
Background
Inflammation is a defense against external stimuli and is manifested by redness, swelling, heat, pain, and dysfunction. Inflammation has been associated with us, partly because of wounds and infections, and partly because of the physiological distress and physical damage it causes.
Inflammation beyond routine may exacerbate the development of various diseases, such as rheumatoid arthritis, Crohn's disease, atherosclerosis, sepsis and tumors, which are all associated with considerable inflammation. The specific expression is the increase of proinflammatory factors, the increase of expression of costimulatory molecules and the like.
In today's society, more and more natural plant based products have been used as the basis for therapeutic human medicine. Although western medicine has become the leading edge of clinical practice today, natural plant products are still used globally for medicine. It is estimated that 80% of the people in developing countries rely mainly on natural products to meet their medical needs. Even in the united states, one in three uses natural drugs. It is estimated that 877 small molecule drugs were introduced globally between 1981 and 2002, of which about 61% are traceable to natural product sources. The natural product is not only effective, but also relatively nontoxic, and the therapeutic dose is far lower than the toxic level.
Isoquinoline alkaloids are also called benzylisoquinoline alkaloids, and have isoquinoline skeleton. Is the biggest alkaloid, and has different classifications according to different connection modes. The medicine is widely applied clinically, most commonly is berberine hydrochloride type, which is extracted from coptis chinensis, phellodendron amurense, barberry root and other plants and can play a role in tranquilizing and relieving pain. The medicine is generally used for resisting tumor, resisting bacteria, relieving pain, regulating immunologic function, resisting platelet aggregation and the like in clinic.
Chinese patent application CN106497875A discloses an application of stephanine and a culture medium and method for amplifying placenta hematopoietic stem cells, which uses stephanine for the preparation of the culture medium, can better culture the hematopoietic stem cells, improve the amplification effect, and has the characteristics of strong dryness and high activity. However, the patent uses stephanine in the culture medium and does not develop its application in other fields.
In conclusion, the application field of stephanine in the prior art is relatively limited, and the anti-inflammatory drugs easily cause stimulation reaction. Based on the above, the isoquinoline alkaloid compounds extracted from Stephania delavayi Diels have good anti-inflammatory activity, and the possibility is provided for developing a new anti-inflammatory drug.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides an anti-inflammatory pharmaceutical composition containing stephanine and a preparation method thereof. The anti-inflammatory pharmaceutical composition provided by the invention contains stephanine, and the stephanine is used in the field of medicine for the first time and does not produce stimulation to human bodies.
In order to achieve the purpose, the invention adopts the technical scheme that:
an anti-inflammatory pharmaceutical composition containing stephanine comprises the following components in percentage by weight: 1-2% of rabdosia rubescens extract, 1-2% of andrographis paniculata extract, 2-3% of cepharanthine, 1-3% of glucose, 0.2-0.4% of essential oil, 0.1-0.3% of emulsifier and 89.3-94.7% of purified water.
Preferably, the stephanine-containing anti-inflammatory pharmaceutical composition comprises the following components in percentage by weight: 1.5 percent of rabdosia rubescens extract, 1.5 percent of andrographis paniculata extract, 2.5 percent of cepharanthine, 2 percent of glucose, 0.3 percent of essential oil, 0.2 percent of emulsifier and 92 percent of purified water.
Preferably, the stephanine is extracted from Stephania delavayi Diels, and the specific process is as follows: collecting radix Stephaniae Dillenii, oven drying, pulverizing, sieving with 100 mesh sieve, cold extracting with 2% hydrochloric acid water solution, alkalifying the extractive solution with NaOH, precipitating, adjusting pH to 9, filtering the precipitate, and drying to obtain total alkaloid extract; extracting the total alkaloid extract with acetone under reflux for 3 hr for 2 times, filtering, concentrating, separating by alumina column chromatography, gradient eluting with aqueous methanol, collecting eluate rich in cepharanthine, concentrating, and drying to obtain crude cepharanthine; recrystallizing the crude stephanine product with acetone to obtain the stephanine.
Preferably, the emulsifier is prepared from soy protein and lecithin in a weight ratio of 3-5: 10 to 13.
Preferably, the emulsifier is prepared from soy protein and lecithin in a weight ratio of 4: 11.
The invention also provides a preparation method of the stephanine-containing anti-inflammatory pharmaceutical composition, which comprises the following steps:
s1, heating purified water with the addition of 50% to 40-50 ℃, adding the rabdosia rubescens extract and the common andrographis herb extract, and mixing and stirring uniformly to obtain a mixture I;
s2, cooling the mixture I obtained in the step S1 to 20-30 ℃, adding stephanine and glucose, and mixing and stirring uniformly to obtain a mixture II;
s3, adding the emulsifier, the essential oil and the residual purified water into the mixture II obtained in the step S2, and mixing and stirring uniformly to obtain the perfume.
Preferably, the mixing and stirring conditions in step S1 are stirring at 500-600 rpm for 20-30 min.
Preferably, the mixing and stirring conditions in the step S2 are stirring at a rotation speed of 350-450 rpm for 40-45 min.
Preferably, the mixing and stirring conditions obtained in the step S3 are stirring at a rotation speed of 550-650 rpm for 35-40 min.
In the invention, an isoquinoline alkaloid compound can be extracted from Stephania delavayi Diels, the compound has better anti-inflammatory activity, and further research on the compound shows that stephanine has an anti-inflammatory effect, so that stephanine is further used for preparing anti-inflammatory drugs.
The inventor uses a proper amount of stephanine in the anti-inflammatory drug, which can obviously improve the anti-inflammatory effect, and in order to reduce the irritation of the anti-inflammatory drug when preparing the pharmaceutical composition, the inventor does not add a preservative and replaces the preservative with essential oil.
Meanwhile, the applicant uses the cepharanthine, the rabdosia rubescens extract and the andrographis paniculata extract together, and adds the cepharanthine, the rabdosia rubescens extract and the andrographis paniculata extract into the medicine according to a certain proportion, so that the anti-inflammatory effect can be obviously improved, the cost of the components is low, the preparation process of the medicine composition is simple, the medicine composition is suitable for being used by the broad masses, the application of the cepharanthine in the anti-inflammatory medicine is realized, and the possibility is provided for developing a new.
Compared with the prior art, the stephanine-containing anti-inflammatory pharmaceutical composition provided by the invention has the following advantages:
(1) according to the stephanine-containing anti-inflammatory pharmaceutical composition provided by the invention, the stephanine, the rabdosia rubescens extract and the andrographis paniculata extract are mixed and added, so that the anti-inflammatory effect is obvious;
(2) according to the stephanine-containing anti-inflammatory pharmaceutical composition provided by the invention, the essential oil and the stephanine are mixed for use, so that the antiseptic effect of the anti-inflammatory drug can be improved, and the stimulation effect of the drug on a human body can be greatly reduced;
(3) the stephanine-containing anti-inflammatory pharmaceutical composition provided by the invention is low in cost and simple in preparation process, and provides possibility for developing a new anti-inflammatory drug.
Detailed Description
The present invention is further explained with reference to the following specific examples, but it should be noted that the following examples are only illustrative of the present invention and should not be construed as limiting the present invention, and all technical solutions similar or equivalent to the present invention are within the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.
The Rabdosia rubescens extract is available from West-Anqiancao biotech GmbH; the Andrographis paniculata Nees extract is available from Hubei Longfei Biotech, Inc.; the essential oil can be obtained from Dongguan cottage commercial and trade Co. Example 1 an anti-inflammatory pharmaceutical composition containing stephanine
The stephanine-containing anti-inflammatory pharmaceutical composition comprises the following components in percentage by weight: 1% of rabdosia rubescens extract, 1% of andrographis paniculata extract, 2% of cepharanthine, 1% of glucose, 0.2% of essential oil, 0.1% of emulsifier and 94.7% of purified water; the stephanine is extracted from Stephania delavayi Diels, and the specific process comprises the following steps: collecting radix Stephaniae Dillenii, oven drying, pulverizing, sieving with 100 mesh sieve, cold extracting with 2% hydrochloric acid water solution, alkalifying the extractive solution with NaOH, precipitating, adjusting pH to 9, filtering the precipitate, and drying to obtain total alkaloid extract; extracting the total alkaloid extract with acetone under reflux for 3 hr for 2 times, filtering, concentrating, separating by alumina column chromatography, gradient eluting with aqueous methanol, collecting eluate rich in cepharanthine, concentrating, and drying to obtain crude cepharanthine; recrystallizing the crude stephanine product with acetone to obtain the final product; the emulsifier is prepared from soybean protein and lecithin in a weight ratio of 3: 10.
The preparation method of the stephanine-containing anti-inflammatory pharmaceutical composition comprises the following steps:
s1, heating purified water with the addition of 50% to 40 ℃, adding the rabdosia rubescens extract and the andrographis paniculata extract, stirring at 500rpm for 20min, and uniformly mixing to obtain a mixture I;
s2, cooling the mixture I obtained in the step S1 to 20 ℃, adding stephanine and glucose, stirring at the rotating speed of 350rpm for 40min, and uniformly mixing to obtain a mixture II;
s3, adding the emulsifier, the essential oil and the residual purified water into the mixture II obtained in the step S2, stirring for 35min at the rotation speed of 550rpm, and uniformly mixing to obtain the oil.
Example 2 an anti-inflammatory pharmaceutical composition containing stephanine
The stephanine-containing anti-inflammatory pharmaceutical composition comprises the following components in percentage by weight: 2% of rabdosia rubescens extract, 2% of common andrographis herb extract, 3% of stephanine, 3% of glucose, 0.4% of essential oil, 0.3% of emulsifier and 89.3% of purified water; the preparation process of the stephanine is similar to that of example 1; the emulsifier is prepared from soybean protein and lecithin in a weight ratio of 5: 13 of the composition.
The preparation method of the stephanine-containing anti-inflammatory pharmaceutical composition comprises the following steps:
s1, heating purified water with the addition of 50% to 50 ℃, adding the rabdosia rubescens extract and the andrographis paniculata extract, stirring at 600rpm for 30min, and uniformly mixing to obtain a mixture I;
s2, cooling the mixture I obtained in the step S1 to 30 ℃, adding stephanine and glucose, stirring for 45min at the rotating speed of 450rpm, and uniformly mixing to obtain a mixture II;
s3, adding the emulsifier, the essential oil and the residual purified water into the mixture II obtained in the step S2, stirring for 38min at the rotating speed of 600rpm, and uniformly mixing to obtain the oil.
Example 3 an anti-inflammatory pharmaceutical composition containing stephanine
The stephanine-containing anti-inflammatory pharmaceutical composition comprises the following components in percentage by weight: 1.5 percent of rabdosia rubescens extract, 1.5 percent of common andrographis herb extract, 2.5 percent of cepharanthine, 2 percent of glucose, 0.3 percent of essential oil, 0.2 percent of emulsifier and 92 percent of purified water; the preparation process of the stephanine is similar to that of example 1; the emulsifier is prepared from soy protein and lecithin in a weight ratio of 4: 11.
The preparation method of the stephanine-containing anti-inflammatory pharmaceutical composition comprises the following steps:
s1, heating purified water with the addition of 50% to 45 ℃, adding the rabdosia rubescens extract and the andrographis paniculata extract, stirring at 550rpm for 25min, and uniformly mixing to obtain a mixture I;
s2, cooling the mixture I obtained in the step S1 to 25 ℃, adding stephanine and glucose, stirring at the rotating speed of 400rpm for 42min, and uniformly mixing to obtain a mixture II;
s3, adding the emulsifier, the essential oil and the residual purified water into the mixture II obtained in the step S2, stirring for 38min at the rotating speed of 600rpm, and mixing and stirring uniformly to obtain the compound.
Comparative example 1 an anti-inflammatory pharmaceutical composition containing stephanine
The stephanine-containing anti-inflammatory pharmaceutical composition comprises the following components in percentage by weight: 1.5 percent of rabdosia rubescens extract, 1.5 percent of common andrographis herb extract, 2.5 percent of cepharanthine, 2 percent of glucose, 0.3 percent of essential oil, 0.2 percent of emulsifier and 92 percent of purified water; the preparation process of the stephanine is similar to that of example 1; the emulsifier is prepared from soybean protein and lecithin in a weight ratio of 1: 1.
The preparation method of the stephanine-containing anti-inflammatory pharmaceutical composition is similar to that of example 3;
the difference from example 3 is that the emulsifier in comparative example 1 is prepared from soy protein and lecithin in a weight ratio of 1: 1.
Comparative example 2 an anti-inflammatory pharmaceutical composition containing stephanine
The stephanine-containing anti-inflammatory pharmaceutical composition comprises the following components in percentage by weight: 1.5 percent of rabdosia rubescens extract, 1.5 percent of common andrographis herb extract, 2 percent of glucose, 0.3 percent of essential oil, 0.2 percent of emulsifier and 94.5 percent of purified water; the preparation process of the stephanine is similar to that of example 1; the emulsifier is prepared from soy protein and lecithin in a weight ratio of 4: 11.
The preparation method of the stephanine-containing anti-inflammatory pharmaceutical composition is similar to that of example 3;
the difference from example 3 is that comparative example 2 does not contain stephanine.
Test example 1 irritation test
1. Test samples: the anti-inflammatory agents prepared in examples 1-3 of the present invention.
2. Test subjects: SPF grade C57BL/6 mice, 60, were evenly divided into 3 groups of 20 mice each.
3. The test method comprises the following steps: before the test, the hair on the two sides of the back spine of the mouse is scraped off without damaging the skin of the mouse, and the hair removal range is 2cm multiplied by 2 cm. 2 μ L of the test sample was applied to one side of the skin with an area of 1.5cm × 1.5cm, covered with two layers of gauze and a layer of cellophane, and fixed with medical tape, and the other side of the skin was treated the same as a blank control. Changes in both sides of the skin were observed at 24h, 48h, respectively.
4. And (3) test results: the final result shows that the skin of the mouse coated with the test sample has no obvious difference from the skin of the mouse not coated with the test sample, which indicates that the anti-inflammatory drugs prepared in the groups of examples 1 to 3 of the invention have no irritation to human body.
Test example 2 Effect of anti-inflammatory drugs on LPS-induced inflammation model of human peripheral blood mononuclear cells
1. Test samples: human peripheral blood cells; gibco serum-free medium; a lipopolysaccharide; dexamethasone; anti-inflammatory drugs obtained in examples 1 to 3 and comparative examples 1 to 2.
2. The test method comprises the following steps: diluting cultured human peripheral blood cells to 200 ten thousand per mL, and inoculatingAdding 200 μ L cell suspension into 96-well plate, placing at 37 deg.C and 5% CO2And (5) incubating in an incubator.
The test was divided into 8 groups, group 1 was blank, and 200. mu.L of LGibco serum-free medium was added; group 2 was LPS group (lipopolysaccharide) to which 100ng/ml lipopolysaccharide was added; the 3 rd group is an anti-inflammatory product dexamethasone experimental group, and anti-inflammatory products 10 mu M are respectively added; 4-8 groups are composed of stephanine-containing anti-inflammatory drug groups, and 2.5 μ M, 5 μ M, and 10 μ M are added respectively; in each group, the drug concentration was repeated in 5 wells. Adding anti-inflammatory drug and LPS, and placing at 37 deg.C and 5% CO2After incubation for 18 hours in the incubator, cell culture supernatants were collected and assayed for the release of the inflammatory factors IL-6, IL-1 β, TNF α by ELISA.
3. And (3) test results: the specific test results are shown in table 1.
TABLE 1 content variation of IL-6, IL-1 β, TNF α in cell culture supernatants
Note: compared with the normal control group,#P<0.05，##P<0.01; comparison with model group*P<0.05，**P<0.01, △ P as compared with the positive control group<0.05；△△P<0.01。
As can be seen from table 1, the anti-inflammatory pharmaceutical compositions of examples 1 to 3 containing cepharanthine according to the present invention have significantly higher anti-inflammatory effects than those of the other groups, while the anti-inflammatory effects of comparative example 2, in which cepharanthine is removed, are greatly reduced, and the anti-inflammatory effects of comparative example 1, in which the components of the emulsifier are changed, are still higher than those of the control group, although the anti-inflammatory effects are reduced.
Test example 3 Effect of the anti-inflammatory agent of the present invention on peritonitis mice
1. Test samples: the stephanine-containing anti-inflammatory pharmaceutical composition prepared in group 3 of example 3 of the present invention.
2. Test animals: SPF grade C57BL/6 80 mice, evenly divided into 8 groups, half male and half female, 8 weeks old [ purchased from nanjing monarch biotechnology limited, quality certification No.: 0017145, animal center license number: SCXK (Su) 2011-.
3. The test method comprises the following steps: injecting LPS 50 mug to the abdominal cavity of a mouse to construct peritonitis; diluting the pharmaceutical composition containing cepharanthine to 50 μ g/mL with physiological saline; the positive control drug was taken as a yankening tablet and treated in the same way (Chongqing company of pharmaceutical industry, national Standard Z20044200).
The 70 mice after the model creation were randomly divided into 7 groups, which were a model group, a positive control group (20mg/kg), and low (5mg/kg), medium (10mg/kg), and high (20mg/kg) dose groups in example 3 of the present invention. And taking another group of mice as a normal control group, and injecting normal saline with the same volume into the abdominal cavity, injecting normal saline with the same volume into the abdominal cavity after model building, and injecting corresponding medicines into the abdominal cavity according to the measurement after medicine experiment building.
4. And (3) test results: the specific test results are shown in Table 2.
TABLE 2 changes in IL-6, IL-1 β, TNF α in the modeled mice
Note: compared with the normal control group,#P<0.05，##P<0.01; comparison with model group*P<0.05，**P<0.01; compared with the positive control group, the test results show that,△P<0.05；△△P<0.01。
as can be seen from table 2, the anti-inflammatory effect of the cepharanthine-containing anti-inflammatory pharmaceutical composition of example 3 of the present invention was different at different concentrations.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (9)
1. An anti-inflammatory pharmaceutical composition containing stephanine is characterized by comprising the following components in percentage by weight: 1-2% of rabdosia rubescens extract, 1-2% of andrographis paniculata extract, 2-3% of cepharanthine, 1-3% of glucose, 0.2-0.4% of essential oil, 0.1-0.3% of emulsifier and 89.3-94.7% of purified water.
2. The stephanine-containing anti-inflammatory pharmaceutical composition according to claim 1, comprising the following components in percentage by weight: 1.5 percent of rabdosia rubescens extract, 1.5 percent of andrographis paniculata extract, 2.5 percent of cepharanthine, 2 percent of glucose, 0.3 percent of essential oil, 0.2 percent of emulsifier and 92 percent of purified water.
3. The stephaganine-containing anti-inflammatory pharmaceutical composition according to claim 1 or 2, wherein said stephaganine is extracted from Stephania delavayi Diels by the process comprising: collecting radix Stephaniae Dillenii, oven drying, pulverizing, sieving with 100 mesh sieve, cold extracting with 2% hydrochloric acid water solution, alkalifying the extractive solution with NaOH, precipitating, adjusting pH to 9, filtering the precipitate, and drying to obtain total alkaloid extract; extracting the total alkaloid extract with acetone under reflux for 3 hr for 2 times, filtering, concentrating, separating by alumina column chromatography, gradient eluting with aqueous methanol, collecting eluate rich in cepharanthine, concentrating, and drying to obtain crude cepharanthine; recrystallizing the crude stephanine product with acetone to obtain the stephanine.
4. The cepharanthine-containing anti-inflammatory pharmaceutical composition according to claim 1 or 2, wherein the emulsifier is composed of soy protein and lecithin in a weight ratio of 3-5: 10 to 13.
5. The cepharanthine-containing anti-inflammatory pharmaceutical composition of claim 4, wherein the emulsifier is composed of soy protein and lecithin in a weight ratio of 4: 11.
6. A process for the preparation of a stephaglabrin-containing anti-inflammatory pharmaceutical composition according to any one of claims 1 to 5, comprising the steps of:
s1, heating purified water with the addition of 50% to 40-50 ℃, adding the rabdosia rubescens extract and the common andrographis herb extract, and mixing and stirring uniformly to obtain a mixture I;
s2, cooling the mixture I obtained in the step S1 to 20-30 ℃, adding stephanine and glucose, and mixing and stirring uniformly to obtain a mixture II;
s3, adding the emulsifier, the essential oil and the residual purified water into the mixture II obtained in the step S2, and mixing and stirring uniformly to obtain the perfume.
7. The method for preparing an anti-inflammatory pharmaceutical composition comprising cepharanthine according to claim 6, wherein the mixing and stirring conditions in step S1 are stirring at 500-600 rpm for 20-30 min.
8. The method for preparing an cepharanthine-containing anti-inflammatory pharmaceutical composition according to claim 6, wherein the mixing and stirring conditions in step S2 are 350 to 450rpm for 40 to 45 min.
9. The method for preparing an cepharanthine-containing anti-inflammatory pharmaceutical composition according to claim 6, wherein the mixing and stirring conditions obtained in step S3 are stirring at 550 to 650rpm for 35 to 40 min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115105502A (en) * | 2022-06-27 | 2022-09-27 | 陕西秦岭七药协同创新中心有限公司 | Application of compound containing stephania plant alkaloid in preparation of feline infectious peritonitis medicine |
| CN116253742A (en) * | 2023-02-21 | 2023-06-13 | 南京工业大学 | Cepharanthine crystal and preparation method thereof |
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| CN115105502A (en) * | 2022-06-27 | 2022-09-27 | 陕西秦岭七药协同创新中心有限公司 | Application of compound containing stephania plant alkaloid in preparation of feline infectious peritonitis medicine |
| CN115105502B (en) * | 2022-06-27 | 2023-08-22 | 陕西秦岭七药协同创新中心有限公司 | Application of compound containing stephania plant alkaloid in preparation of cat infectious peritonitis medicine |
| CN116253742A (en) * | 2023-02-21 | 2023-06-13 | 南京工业大学 | Cepharanthine crystal and preparation method thereof |
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