Polar part of pharmaceutical composition as well as preparation method and application of polar part

Abstract

The invention belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to a polar part of a pharmaceutical composition as well as a preparation method and application thereof. The preparation method of the polar part of the pharmaceutical composition comprises the following steps: concentrating the prepared oral liquid, and respectively and sequentially extracting the petroleum ether part extract, the chloroform part extract, the ethyl acetate part freeze-dried powder, the n-butyl alcohol part freeze-dried powder and the water part freeze-dried powder; respectively adding the petroleum ether part extract, the chloroform part extract, the ethyl acetate part freeze-dried powder, the n-butyl alcohol part freeze-dried powder and the water part freeze-dried powder into DMSO (Dimethylsulfoxide) for dissolving, and fixing the volume. The polar part prepared by the preparation method of the polar part provided by the invention has good anti-inflammatory, antipyretic and analgesic effects.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine extraction technology, specifically relating to a polar fraction of a pharmaceutical composition, its preparation method, and its application. Background Technology

[0002] Common childhood cold due to wind-heat is a frequent pediatric illness, mainly characterized by high fever, slight aversion to wind, headache, sore throat, dry mouth, and nasal congestion with yellow discharge. Traditional Chinese medicine theory holds that children's internal organs are delicate, their constitution is not fully developed, and their defensive functions are weak, making them susceptible to external pathogens. Guided by this theory, treatment formulas for childhood wind-heat cold have emerged. This oral liquid formulation is a traditional Chinese medicine preparation developed based on this theory and combined with modern pharmaceutical technology. Its formula typically includes various Chinese herbal ingredients such as honeysuckle, forsythia, isatis root, peppermint, schizonepeta, gypsum, and anemarrhena, possessing the effects of relieving exterior syndromes with pungent and cooling properties, and clearing heat and detoxifying. Traditional Chinese medicine dosage forms are mostly decoctions, requiring preparation before administration, which is inconvenient for children. The advent of oral liquid preparations not only preserves the efficacy of traditional Chinese medicine but also offers advantages such as convenient administration, rapid absorption, and accurate dosage, making them particularly suitable for children. The development of modern pharmaceutical technology, such as the application of supercritical fluid extraction and membrane separation, has further improved the content and bioavailability of active ingredients in oral liquid drug compositions, while reducing the risk of adverse reactions. However, problems such as low bioavailability and low efficacy still exist.

[0003] Relevant patent documents retrieved: For example, Chinese patent CN102600378A, published on January 22, 2014, discloses a method for preparing a pharmaceutical composition for treating pediatric colds using ultrafiltration. First, volatile oils are extracted from honeysuckle, forsythia, schizonepeta, peppermint, immature bitter orange, and bupleurum from the raw materials. The distilled aqueous solution is collected separately. The residue is decocted with the remaining fourteen raw materials, filtered, and the filtrate is concentrated. The concentrated solution is then centrifuged at high speed. The centrifuged solution is ultrafiltered, and finally, the extracted volatile oils and distilled aqueous solution are added to the ultrafiltrate. The mixture is filtered, filled, sterilized, and then conventional excipients are added. Following conventional processes, various clinically acceptable formulations are produced. This preparation method uses modern ultrafiltration purification technology to replace alcohol precipitation, which can retain ineffective large molecules while preserving effective small biological molecules. This improves the clarity of the liquid, eliminates ethanol residue, ensures the product meets standard requirements, and guarantees the safety and efficacy of the medication for children.

[0004] Relevant non-patent literature retrieved: The journal or book title is *Chinese Journal of Traditional Chinese Medicine*, and the article title is "Analysis of Differences in Active Components and Antioxidant Activity of Different Polar Fractions of Honeysuckle Alcohol Extract," volume number 2022.11.045. This study investigated the composition of active components and differences in antioxidant activity of honeysuckle alcohol extract and its different polar fractions. The honeysuckle alcohol extract (M) was divided into water (M5), n-butanol (M4), ethyl acetate (M3), dichloromethane (M2), and petroleum ether (M1) fractions. Qualitative and quantitative analyses of the active components in each fraction were performed, and their antioxidant activities were compared. The main active component compositions of M4, M5, and M were basically the same. However, among the different polar fractions, M5 had the highest contents of cryptochlorogenic acid, loganic acid, and neochlorogenic acid; M4 had the highest contents of chlorogenic acid, loganin, oxidized loganin, rutin, and luteolin; M3 had the highest contents of isochlorogenic acids A, B, and C; and the contents of active substances in M2 and M1 were relatively low. The results showed that the main components of ethyl acetate, which has the highest antioxidant activity, are isochlorogenic acids A, B, and C, which have the potential to become natural antioxidants, providing a certain reference for further in-depth research, development, and utilization of honeysuckle resources.

[0005] The prior art represented by the aforementioned documents has at least the following unresolved technical problems or defects: The aforementioned literature still suffers from problems such as low bioavailability and low efficacy. Summary of the Invention

[0006] The purpose of this invention is to provide: a polar site of a pharmaceutical composition, its preparation method and application, and related technologies, to solve technical problems such as high impurity content, low density, large crystal size, or combinations thereof.

[0007] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0008] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0009] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0010] In a first aspect, the present invention provides a method for preparing the polar portion of a pharmaceutical composition, wherein the pharmaceutical composition comprises, by weight parts, the following raw materials: 15-40 parts of honeysuckle, 70-90 parts of forsythia, 20-30 parts of isatis root, 13-34 parts of peppermint, 15-25 parts of bupleurum, 41-60 parts of burdock fruit, 9-13 parts of schizonepeta spike, 80-95 parts of gypsum, 55-80 parts of scutellaria, 20-40 parts of gardenia, 3-8 parts of platycodon, 20-35 parts of red peony root, 10-18 parts of reed rhizome, 17-25 parts of stir-fried bitter almond, 17-30 parts of light bamboo leaf, 15-32 parts of immature bitter orange peel, 5-12 parts of saposhnikovia root, 10-29 parts of stir-fried medicated leaven, 1-4 parts of silkworm pupa, and 28-45 parts of licorice. The method for preparing the polar portion of the pharmaceutical composition includes the following steps: (1) Soak honeysuckle, forsythia, schizonepeta, mint, bitter orange peel, and bupleurum in water, then distill them. Collect the volatile oil and the distilled aqueous solution separately for later use. Boil the distilled residue and the remaining 14 herbs in water, filter, and obtain filtrate 1 and residue 1. Boil residue 1 in water again, filter, and obtain filtrate 2 and residue 2. Combine filtrate 1 and filtrate 2, filter, and concentrate to obtain clear extract 1. (2) Add ethanol to the clear paste 1 for alcohol precipitation. After the alcohol precipitation is completed, filter and concentrate the filtrate to obtain clear paste 2. Add the volatile oil obtained in step (1) and the distilled aqueous solution to clear paste 2, mix well, add water to adjust the density, and obtain the mixed liquid preparation. (3) Sterilize the mixed liquid preparation to obtain an oral liquid, concentrate the oral liquid, and extract the petroleum ether fraction extract, chloroform fraction extract, ethyl acetate fraction lyophilized powder, n-butanol fraction lyophilized powder and water fraction lyophilized powder in sequence. (4) Dissolve the petroleum ether extract, chloroform extract, ethyl acetate lyophilized powder, n-butanol lyophilized powder, and water lyophilized powder in DMSO and bring the volume to a final volume.

[0011] Preferably, the pharmaceutical composition comprises the following raw materials in parts by weight: 40 parts honeysuckle, 70 parts forsythia, 20 parts isatis root, 34 parts peppermint, 17 parts bupleurum, 48 parts burdock fruit, 10 parts schizonepeta spike, 90 parts gypsum, 63 parts scutellaria, 20 parts gardenia, 8 parts platycodon, 35 parts red peony root, 15 parts reed rhizome, 18 parts stir-fried bitter almond, 20 parts light bamboo leaf, 32 parts immature bitter orange peel, 10 parts saposhnikovia root, 15 parts stir-fried medicated leaven, 4 parts silkworm, and 32 parts licorice.

[0012] Preferably, in step (1), the ratio of raw material to water for soaking is 1g:5-8mL, and the soaking time is 20-40min.

[0013] More preferably, in step (1), the ratio of raw material to water for soaking is 1g:6mL, and the soaking time is 30min.

[0014] Preferably, the total weight of the distilled residue and the remaining 14 herbs in step (1) is 1g:5-8mL with the volume of water added. The decoction is prepared for 1.3-2 hours and filtered through 8 layers of gauze.

[0015] More preferably, in step (1), the total weight of the distilled residue and the remaining 14 herbs in step (1) is 1g:6mL, the decoction is prepared for 1.5h, and then filtered through 8 layers of gauze.

[0016] Preferably, the ratio of filter residue 1 to water in step (1) is 1g:4-5mL, and the mixture is boiled for 1-2 hours and filtered through 8 layers of gauze.

[0017] More preferably, in step (1), the ratio of filter residue 1 to water is 1g:4mL, the mixture is decocted for 1 hour, and then filtered through 8 layers of gauze.

[0018] Preferably, the degree of concentration in step (1) is to concentrate to a clear paste with a relative density of 1-2, and the temperature of the clear paste is 55-60℃.

[0019] More preferably, the degree of concentration in step (1) is to concentrate to a clear paste with a relative density of 1.12.

[0020] Preferably, the volume fraction of ethanol in step (2) is 90-96%, and the addition of ethanol is stopped when the alcohol content of the supernatant reaches 70%.

[0021] More preferably, the volume fraction of ethanol in step (2) is 95%.

[0022] Preferably, the degree of concentration in step (2) is to concentrate to a clear paste with a relative density of 1-2.

[0023] More preferably, the degree of concentration in step (2) is to concentrate to a clear paste with a relative density of 1.15.

[0024] Preferably, step (2) involves adjusting the density to 1.01 (relative to the density of water).

[0025] Preferably, the degree of concentration in step (3) is to concentrate to 1 / 3 of the oral liquid volume.

[0026] Preferably, the preparation method of the petroleum ether fraction extract in step (3) includes the following steps: adding petroleum ether to the concentrated oral liquid, shaking and allowing it to stand to separate into layers, collecting the petroleum ether layer, repeating the extraction, combining the extracts, concentrating, and obtaining the petroleum ether fraction extract.

[0027] More preferably, the preparation method of the petroleum ether fraction extract in step (3) includes the following steps: take 100 mL of the concentrated oral liquid, add 100 mL of petroleum ether, shake thoroughly and let stand to separate the layers, collect the petroleum ether layer; repeat the extraction 4 times until the petroleum ether layer is colorless, combine the extracts, concentrate under reduced pressure (40-50°C) to remove the petroleum ether, and obtain the petroleum ether fraction extract.

[0028] Preferably, the method for preparing the chloroform fraction extract in step (3) includes the following steps: discarding the petroleum ether layer, adding chloroform to the aqueous layer, repeating the extraction, combining the organic layers, and concentrating to obtain the chloroform fraction extract.

[0029] More preferably, the method for preparing the chloroform fraction extract in step (3) includes the following steps: after discarding the petroleum ether layer, 100 mL of chloroform is added to the aqueous layer, the extraction is repeated 4 times, the organic layers are combined, and the chloroform is concentrated under reduced pressure to obtain the chloroform fraction extract.

[0030] Preferably, the method for preparing the ethyl acetate lyophilized powder in step (3) includes the following steps: discarding the chloroform layer, adding ethyl acetate to the aqueous layer, repeating the extraction, combining the organic layers, evaporating, and lyophilizing to obtain the ethyl acetate lyophilized powder.

[0031] More preferably, the method for preparing the ethyl acetate lyophilized powder in step (3) includes the following steps: after discarding the chloroform layer, 100 mL of ethyl acetate is added to the aqueous layer, the extraction is repeated 4 times, the organic layers are combined, most of the solvent is initially removed by a rotary evaporator, the remaining solution is transferred to a lyophilization bottle, and lyophilized using a freeze dryer (-50°C, under vacuum) to obtain the ethyl acetate lyophilized powder.

[0032] Preferably, the method for preparing the lyophilized n-butanol fraction in step (3) includes the following steps: discarding the ethyl acetate layer, adding n-butanol to the aqueous layer, repeating the extraction, merging the organic layers, evaporating, and lyophilizing to obtain the lyophilized n-butanol fraction.

[0033] More preferably, the method for preparing the lyophilized n-butanol fraction in step (3) includes the following steps: after discarding the ethyl acetate layer, 100 mL of n-butanol is added to the aqueous layer, the extraction is repeated 4 times and the organic layers are combined, the liquid is first concentrated by rotary evaporation to remove most of the liquid, and then freeze-dried to obtain the lyophilized n-butanol fraction.

[0034] Preferably, the method for preparing the water-based freeze-dried powder in step (3) includes the following steps: discarding the n-butanol layer, further concentrating the water layer, and drying it to obtain the water-based freeze-dried powder.

[0035] More preferably, the method for preparing the water fraction freeze-dried powder in step (3) includes the following steps: after discarding the n-butanol layer, the water layer is further concentrated to 50 mL and directly freeze-dried to obtain the water fraction freeze-dried powder.

[0036] Preferably, the mass fraction of DMSO in step (4) is 4-6%.

[0037] More preferably, the mass fraction of DMSO in step (4) is 5%.

[0038] Secondly, the present invention provides a polar portion of a pharmaceutical composition prepared by the above-described preparation method.

[0039] Thirdly, the present invention provides the application of the polar portion of the pharmaceutical composition obtained by the above preparation method in the preparation of anti-inflammatory, antipyretic, and analgesic drugs.

[0040] The present invention has the following beneficial effects: The polar fractions of the pharmaceutical composition obtained by this invention exhibit good anti-inflammatory, antipyretic, and analgesic effects. The anti-inflammatory effects, from strongest to weakest, are: ethyl acetate fraction > aqueous fraction > petroleum ether fraction > n-butanol fraction > chloroform fraction; the antipyretic effects, from strongest to weakest, are: ethyl acetate fraction > n-butanol fraction > petroleum ether fraction > aqueous fraction > chloroform fraction; the peripheral analgesic effects, from strongest to weakest, are: petroleum ether > aqueous fraction > chloroform fraction > n-butanol fraction > ethyl acetate fraction; and the central analgesic effects, from strongest to weakest, are: aqueous fraction > n-butanol fraction > ethyl acetate fraction > petroleum ether fraction > chloroform fraction. Attached Figure Description

[0041] Figure 1 The effect of different polar fractions of the drug composition on xylene-induced ear swelling in mice (where ** represents p < 0.01 compared with the model group, *** represents p < 0.001 compared with the model group, and ns represents no significant difference compared with the model group). Figure 2 The effect of different polar fractions of the drug composition on xylene-induced IL-1β levels in mouse serum (where * represents p < 0.05 compared to the model group, and ns represents no significant difference compared to the model group). Figure 3 The effect of different polar fractions of the drug composition on xylene-induced TNF-α levels in mouse serum (where * represents p < 0.05 compared to the model group, and ns represents no significant difference compared to the model group). Figure 4 The diagram shows the effect of different polar portions of the drug composition on the interaction effect of body temperature in rats (where * represents p<0.05 compared with the model group, and ** represents p<0.01 compared with the model group). Figure 5 The effect of different polar portions of the drug composition on the number of writhing movements in mice (where * represents p<0.05 compared with the model group, ** represents p<0.01 compared with the model group, **** represents p<0.0001 compared with the model group, and ns represents no significant difference compared with the model group). Figure 6 A profile of the effect of different polar portions of the drug composition on the pain threshold in mice (where * represents p<0.05 compared with the control group). Figure 7 The graph shows the anti-inflammatory, antipyretic, and analgesic effects at different polar sites in different embodiments (where * represents p<0.05 compared with the model group, ** represents p<0.01 compared with the model group, *** represents p<0.001 compared with the model group, and **** represents p<0.0001 compared with the model group; in central paroxysmal pain, * represents p<0.05 compared with the control group, and ns represents no significant difference compared with the control group). Figure 8 The graph shows the anti-inflammatory, antipyretic, and analgesic effects at different polarity sites (where * represents p<0.05 compared to the model group, ** represents p<0.01 compared to the model group, *** represents p<0.001 compared to the model group, and ns represents p<0.001 compared to the model group; for central paroxysmal pain, ns represents no significant difference compared to the control group). Detailed Implementation

[0042] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0043] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0044] The pharmaceutical compositions of Examples 1-3, by weight (1 part equals 1 g), consist of the following raw materials (Table 1): Table 1

[0045] The above Examples 1-3 were prepared according to the following method: (1) Add 6 times the amount of ultrapure water to the raw materials of honeysuckle, forsythia, schizonepeta, peppermint, immature bitter orange, and bupleurum, soak for 30 minutes, and extract the volatile oil by steam distillation. During the distillation process, collect the volatile oil and the distilled aqueous solution separately for later use. Mix the distilled residue with the remaining fourteen raw materials (total of all herbs), add 6 times the amount of ultrapure water and decoct. Heat to boiling and continue to decoct for 1.5 hours. Filter the extract through 8 layers of gauze. Add 4 times the amount of ultrapure water to the residue, shake well, heat to boiling and continue to decoct for 1 hour. Filter the extract through 8 layers of gauze. After decoction, combine the decoctions and filter to remove impurities. Concentrate the filtrate to a clear paste with a relative density of 1.12. The temperature of the paste is measured to be 55-60℃.

[0046] (2) Slowly add 95% ethanol to the concentrated and cooled clear paste at room temperature, stir evenly, and stop adding ethanol when the alcohol content of the supernatant reaches 70%. Let it stand for more than 48 hours and carry out alcohol precipitation treatment to further purify the effective components. After the alcohol precipitation is completed, filter and collect the filtrate, and concentrate it again to a clear paste with a relative density of 1.15. The temperature of the paste is measured to be 55-60℃. Add the volatile oil extracted in step (1) and the distilled aqueous solution to the concentrated clear paste, mix thoroughly, and add pure water to adjust the density to 1.01. (3) The mixed liquid preparation was sterilized and finally diluted to 300 mL to obtain an oral liquid preparation (1 mL of the oral liquid preparation is equivalent to 2.0 g of raw medicinal material); the oral liquid preparation was concentrated to 1 / 3 of its original volume (100 mL); the petroleum ether fraction extract, chloroform fraction extract, ethyl acetate fraction lyophilized powder, n-butanol fraction lyophilized powder and water fraction lyophilized powder were extracted in sequence. ① Preparation of petroleum ether fraction extract Take one portion of the concentrated oral solution, add 100 mL of petroleum ether, shake thoroughly, and allow to stand for separation. Collect the petroleum ether layer. Repeat the extraction 4 times until the petroleum ether layer is colorless. Combine the extracts and concentrate under reduced pressure (40-50°C) to remove the petroleum ether, obtaining the petroleum ether fraction extract.

[0047] ②Preparation of chloroform extract After discarding the petroleum ether layer, 100 mL of chloroform was added to the aqueous layer, and the extraction was repeated 4 times. The organic layers were combined, and the chloroform was concentrated under reduced pressure to obtain the chloroform extract.

[0048] ③ Preparation of ethyl acetate extract After discarding the chloroform layer, 100 mL of ethyl acetate was added to the aqueous layer, and the extraction was repeated 4 times. The organic layers were combined, and most of the solvent was removed by rotary evaporation. The remaining solution was transferred to a lyophilization bottle and lyophilized using a freeze dryer (-50°C, under vacuum) to obtain lyophilized powder of the ethyl acetate fraction.

[0049] ④ Preparation of n-Butanol fraction extract After discarding the ethyl acetate layer, 100 mL of n-butanol was added to the aqueous layer. The extraction was repeated 4 times and the organic layers were combined. The liquid was first concentrated by rotary evaporation to remove most of the liquid, and then freeze-dried to obtain the lyophilized n-butanol fraction.

[0050] ⑤ Preparation of aqueous extract After discarding the n-butanol layer, the aqueous layer was further concentrated to 50 mL and then freeze-dried to obtain the water fraction as freeze-dried powder.

[0051] (4) Dissolve the petroleum ether extract, chloroform extract, ethyl acetate lyophilized powder, n-butanol lyophilized powder, and water lyophilized powder separately in 5.0% DMSO, and then make up to 300 mL with ultrapure water to obtain the final product.

[0052] Comparative Example 1 Unlike Example 1, the amount of water added in each step (1) is 3 times the amount of ultrapure water.

[0053] Comparative Example 2 Unlike Example 1, the soaking time in step (1) is 10 minutes, and the time for decocting the distilled residue with the remaining fourteen raw materials is 1 hour.

[0054] Comparative Example 3 Unlike Example 1, in step (2), the addition of alcohol was stopped when the alcohol content of the supernatant reached 65%.

[0055] Comparative Example 4 Unlike Example 1, the alcohol precipitation time in step (2) is 24 hours.

[0056] Comparative Example 5 The difference from Example 1 is that the raw material ratio is as follows: honeysuckle 50 parts, forsythia 50 parts, isatis root 30 parts, peppermint 34 parts, bupleurum 17 parts, burdock fruit 48 parts, schizonepeta spike 10 parts, gypsum 90 parts, scutellaria 63 parts, gardenia 20 parts, platycodon 8 parts, red peony root 35 parts, reed rhizome 15 parts, stir-fried bitter almond 18 parts, light bamboo leaf 20 parts, bitter orange peel 32 parts, saposhnikovia root 10 parts, stir-fried medicated leaven 15 parts, silkworm 4 parts, and licorice 32 parts, totaling 601g (1 part is 1g).

[0057] Comparative Example 6 Unlike Example 1, the raw material ratio is as follows: 40 parts honeysuckle, 70 parts forsythia, 20 parts isatis root, 40 parts peppermint, 30 parts bupleurum, 60 parts burdock fruit, 20 parts schizonepeta spike, 49 parts gypsum, 63 parts scutellaria, 20 parts gardenia, 8 parts platycodon, 35 parts red peony root, 15 parts reed rhizome, 18 parts stir-fried bitter almond, 20 parts light bamboo leaf, 32 parts immature bitter orange peel, 10 parts saposhnikovia root, 15 parts stir-fried medicated leaven, 4 parts silkworm pupa, and 32 parts licorice, totaling 601g (1 part equals 1g).

[0058] Comparative Example 7 Unlike Example 1, the raw material ratio is as follows: 70 parts honeysuckle, 50 parts forsythia, 10 parts isatis root, 49 parts peppermint, 10 parts bupleurum, 30 parts burdock fruit, 20 parts schizonepeta spike, 60 parts gypsum, 50 parts scutellaria, 10 parts gardenia, 20 parts platycodon, 45 parts red peony root, 30 parts reed rhizome, 10 parts stir-fried bitter almond, 10 parts light bamboo leaf, 40 parts immature bitter orange peel, 20 parts saposhnikovia root, 40 parts stir-fried medicated leaven, 7 parts silkworm pupa, and 20 parts licorice, totaling 601g (1 part equals 1g).

[0059] Comparative Example 8 Unlike Example 1, the raw material ratio is as follows: 70 parts honeysuckle, 50 parts forsythia, 30 parts isatis root, 40 parts peppermint, 30 parts bupleurum, 40 parts burdock fruit, 3 parts schizonepeta spike, 90 parts gypsum, 63 parts scutellaria, 20 parts gardenia, 8 parts platycodon, 35 parts red peony root, 15 parts reed rhizome, 18 parts stir-fried bitter almond, 20 parts light bamboo leaf, 32 parts immature bitter orange peel, 10 parts saposhnikovia root, 15 parts stir-fried medicated leaven, 2 parts silkworm pupa, and 10 parts licorice, totaling 601g (1 part equals 1g).

[0060] Effect Experiment Establishment and grouping of animal models for anti-inflammatory, antipyretic, analgesic, and antibacterial effects Rearing conditions: Lighting is set to alternate between light and dark for 12 hours. The cages are kept in a clean laminar flow rack with constant temperature (20-26℃) and constant humidity (50%-65%). The cages, bedding, feed and drinking water are all sterilized by high pressure steam and changed at least once a week under sterile conditions. No more than 5 mice are kept in each cage. 1. Xylene-induced mouse ear swelling model Thirty 6-8 week old SPF-grade male Kunming mice, weighing 18-22 g, were acclimatized for one week. They were randomly divided into four groups (model group, petroleum ether group, chloroform group, ethyl acetate group, n-butanol group, and water group), with five mice in each group. The mice were administered the drug via gavage at a dose of 10.80 mL / kg; the model group received an equal volume of distilled water via gavage. The administration was once daily for five consecutive days. One hour after the last administration, 30 μL of xylene was rapidly injected into the anterior and posterior sides of the left ear (15 μL on each side) using a micropipette to establish an ear swelling model. Forty minutes after inflammation, the eyeballs were enucleated, blood was collected, serum was separated, and stored at -20°C for later use. Both ears were cut open, and symmetrical ear pieces were taken from the left and right ears using an 8 mm punch. The weight difference between the left and right ear pieces was used to determine the degree of swelling. Figure 1-3 As shown, the ear swelling inhibition rates of the drug composition at different polarities, from highest to lowest, are: ethyl acetate fraction > water fraction > petroleum ether fraction > n-butanol fraction > chloroform fraction. Simultaneously, administration to different polarities reduced the serum levels of TNF-α and IL-1β in inflamed mice to varying degrees.

[0061] 2. Dry yeast-induced fever model in rats Eighty rats were acclimatized for one week, and their rectal temperature was measured once daily at a fixed time for three consecutive days. The procedure was as follows: the rats were restrained, and the probe of an electronic thermometer, coated with Vaseline, was inserted 3 cm into the rat's rectum. After the thermometer beeped, the reading was taken for 40 seconds to allow the rats to acclimatize to the experimental conditions and reduce stress caused by temperature measurement. Seventy rats whose body temperature remained between 36.5-38.3℃ for two consecutive days, with a variation not exceeding 0.5℃, were selected, and the average of the two-day temperature was used as the rat's basal body temperature.

[0062] Fifty-six selected rats were randomly divided into four groups according to body weight: a control group, a model group, a petroleum ether group, a chloroform group, an ethyl acetate group, a n-butanol group, and a water group, with eight rats in each group. Rats were administered the drug via gavage at a dose of 7.5 mL / kg; the control and model groups were administered an equal volume of distilled water via gavage. On the third day after gavage, except for the control group, all rats in each group were subcutaneously injected with 15% dry yeast suspension (10 mL / kg) in their backs to establish a fever model. Body temperature was measured every 1 hour after model establishment, for a total of 10 measurements. Results are as follows: Figure 4 The antipyretic effects of different polarities of the drug composition, from greatest to least, are as follows: ethyl acetate fraction > n-butanol fraction > petroleum ether fraction > water fraction > chloroform fraction.

[0063] 3. Mouse model of painful writhing induced by glacial acetic acid Thirty SPF-grade male Kunming mice, weighing 18-22 g, were acclimatized for one week and randomly divided into four groups (n=5 per group): a model group, a petroleum ether group, a chloroform group, an ethyl acetate group, a n-butanol group, and a water group. Mice were administered the drug via gavage at a dose of 10.80 mL / kg, while the model group received an equal volume of distilled water via gavage. This was repeated once daily for five consecutive days. One hour after the last administration, all mice in each group were intraperitoneally injected with 0.6% glacial acetic acid at a dose of 10 mL / kg. The number of writhing movements (abdominal contraction, body twisting, hind limb and trunk extension, and buttock elevation) in each group was recorded within 5-30 minutes after the acetic acid injection. Results are as follows: Figure 5 The peripheral analgesic effects of the drug composition at different polarities, from greatest to least, are: petroleum ether > water > chloroform > n-butanol > ethyl acetate.

[0064] 4. Mouse hot plate pain model Sixty SPF-grade female Kunming mice, weighing 18-22 g, were acclimatized for one week and placed on an aluminum plate in a 55.0±0.5℃ water bath. The time it took for each mouse to lick or raise its hind paw after being placed on the hot plate was recorded using a stopwatch as its pain threshold. The pain threshold was measured again after a 5-minute interval. Forty-eight mice with stable pain threshold responses (not frequently jumping) between 5-30 seconds were selected. These 48 mice were randomly divided into four groups (n=8 per group): a control group, a petroleum ether group, a chloroform group, an ethyl acetate group, a n-butanol group, and a water group. The mice were administered the drug via gavage at a dose of 10.80 mL / kg, while the control group received an equal volume of distilled water via gavage. The administration was once daily for five consecutive days. At 30, 60, 90, and 120 min after the last administration, mice were placed on aluminum plates preheated to (55.1±0.1)℃, and their reaction time to licking or raising their hind paws was observed. The pain threshold of each mouse was measured, and if the pain threshold exceeded 60 s, it was counted as 60 s. The results are as follows: Figure 6 The central analgesic effects of different polarity sites of the drug composition, from greatest to least, are as follows: water site > n-butanol site > ethyl acetate site > petroleum ether site > chloroform site.

[0065] The results above show that the effects of different polar fractions of the drug composition are as follows: anti-inflammatory effect from largest to smallest: ethyl acetate fraction > water fraction > petroleum ether fraction > n-butanol fraction > chloroform fraction; antipyretic effect from largest to smallest: ethyl acetate fraction > n-butanol fraction > petroleum ether fraction > water fraction > chloroform fraction; peripheral analgesic effect from largest to smallest: petroleum ether > water > chloroform > n-butanol > ethyl acetate fraction; central analgesic effect from largest to smallest: water fraction > n-butanol fraction > ethyl acetate fraction > petroleum ether fraction > chloroform fraction. Based on the anti-inflammatory, antipyretic, and analgesic effects of different polar fractions of the pharmaceutical composition, the fractions with the best effects were selected for comparison in different embodiments and comparative examples. This included comparing the anti-inflammatory and antipyretic effects of the ethyl acetate fraction extract, the peripheral analgesic effects of the petroleum ether fraction, and the central analgesic effects of the water fraction. Figure 7 and 8 As shown.

[0066] Depend on Figure 7 It can be seen that the drugs prepared in Examples 1-3 have significant anti-inflammatory, antipyretic, and analgesic effects. Specifically, the anti-inflammatory effect is significant; Examples 1-3 can significantly inhibit xylene-induced ear swelling, reducing the ear swelling degree from about 27mg in the model group to an average of 10-13mg (p<0.001); the antipyretic effect is significant; in the fever model, Examples 1-3 significantly inhibited the rise in body temperature, with Example 1 showing the best effect; the analgesic effect is outstanding; the petroleum ether extract of Examples 1-3 can significantly relieve peripheral pain, and the aqueous extract of Example 1 can effectively relieve central agonistic pain.

[0067] Depend on Figure 8 It can be seen that the comparative samples 1-8 have certain anti-inflammatory, antipyretic, and analgesic effects, but the effects are not obvious, especially the antipyretic effect, in which no significant difference was shown in the comparative samples 3-8; the central analgesic effect also showed no significant difference in the comparative samples 1-8.

[0068] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing the polar portion of a pharmaceutical composition, characterized in that, The pharmaceutical composition comprises, by weight, the following raw materials: honeysuckle 15-40 parts, forsythia 70-90 parts, isatis root 20-30 parts, peppermint 13-34 parts, bupleurum 15-25 parts, burdock fruit 41-60 parts, schizonepeta spike 9-13 parts, gypsum 80-95 parts, scutellaria baicalensis 55-80 parts, gardenia 20-40 parts, platycodon root 3-8 parts, red peony root 20-35 parts, reed rhizome 10-18 parts, stir-fried bitter almond 17-25 parts, light bamboo leaf 17-30 parts, immature bitter orange peel 15-32 parts, saposhnikovia root 5-12 parts, stir-fried medicated leaven 10-29 parts, silkworm pupa 1-4 parts, and licorice root 28-45 parts; The method for preparing the polar portion of the pharmaceutical composition includes the following steps: (1) Soak honeysuckle, forsythia, schizonepeta, mint, bitter orange peel, and bupleurum in water, then distill them. Collect the volatile oil and the distilled aqueous solution separately for later use. Boil the distilled residue and the remaining 14 herbs in water, filter, and obtain filtrate 1 and residue 1. Boil residue 1 in water again, filter, and obtain filtrate 2 and residue 2. Combine filtrate 1 and filtrate 2, filter, and concentrate to obtain clear extract 1. (2) Add ethanol to the clear paste 1 for alcohol precipitation. After the alcohol precipitation is completed, filter and concentrate the filtrate to obtain clear paste 2. Add the volatile oil obtained in step (1) and the distilled aqueous solution to clear paste 2, mix well, add water to adjust the density, and obtain the mixed liquid preparation. (3) Sterilize the mixed liquid preparation, make up the volume to obtain an oral liquid, concentrate the oral liquid, and extract the petroleum ether fraction extract, chloroform fraction extract, ethyl acetate fraction lyophilized powder, n-butanol fraction lyophilized powder and water fraction lyophilized powder in sequence. (4) Dissolve the petroleum ether extract, chloroform extract, ethyl acetate lyophilized powder, n-butanol lyophilized powder, and water lyophilized powder in DMSO and bring the volume to a final volume.

2. The preparation method according to claim 1, characterized in that, The pharmaceutical composition comprises, by weight, the following ingredients: 40 parts honeysuckle, 70 parts forsythia, 20 parts isatis root, 34 parts peppermint, 17 parts bupleurum, 48 parts burdock fruit, 10 parts schizonepeta spike, 90 parts gypsum, 63 parts scutellaria, 20 parts gardenia, 8 parts platycodon, 35 parts red peony root, 15 parts reed rhizome, 18 parts stir-fried bitter almond, 20 parts light bamboo leaf, 32 parts immature bitter orange peel, 10 parts saposhnikovia root, 15 parts stir-fried medicated leaven, 4 parts silkworm pupa, and 32 parts licorice.

3. The preparation method according to claim 1, characterized in that, In step (1), the ratio of raw material to water for soaking is 1g:5-8mL, and the soaking time is 20-40min.

4. The preparation method according to claim 1, characterized in that, The total weight of the distilled residue and the remaining 14 herbs in step (1) is 1g:5-8mL with the volume of water added. The herbs are decocted for 1.3-2 hours and filtered through 8 layers of gauze. The ratio of the filter residue 1 in step (1) to the liquid with water added is 1g:4-5mL. The herbs are decocted for 1-2 hours and filtered through 8 layers of gauze.

5. The preparation method according to claim 1, characterized in that, The degree of concentration mentioned in step (1) is to concentrate to a clear paste with a relative density of 1-2, and the temperature of the clear paste is 55-60℃.

6. The preparation method according to claim 1, characterized in that, The volume fraction of ethanol in step (2) is 90-96%, and the addition of ethanol is stopped when the alcohol content of the supernatant reaches 70%.

7. The preparation method according to claim 1, characterized in that, The preparation method of the petroleum ether fraction extract described in step (3) includes the following steps: adding petroleum ether to the concentrated oral liquid, shaking and allowing it to stand to separate into layers, collecting the petroleum ether layer, repeating the extraction, combining the extracts, concentrating, and obtaining the petroleum ether fraction extract. The method for preparing the chloroform fraction extract includes the following steps: discarding the petroleum ether layer, adding chloroform to the aqueous layer, repeating the extraction, combining the organic layers, and concentrating to obtain the chloroform fraction extract; The method for preparing the ethyl acetate lyophilized powder includes the following steps: discarding the chloroform layer, adding ethyl acetate to the aqueous layer, repeating the extraction, combining the organic layers, evaporating, and lyophilizing to obtain the ethyl acetate lyophilized powder. The method for preparing the lyophilized n-butanol fraction includes the following steps: discarding the ethyl acetate layer, adding n-butanol to the aqueous layer, repeating the extraction, combining the organic layers, evaporating, and lyophilizing to obtain the lyophilized n-butanol fraction. The method for preparing the aqueous freeze-dried powder includes the following steps: discarding the n-butanol layer, further concentrating the aqueous layer, and drying it to obtain the aqueous freeze-dried powder.

8. The preparation method according to claim 1, characterized in that, The mass fraction of DMSO mentioned in step (4) is 4-6%.

9. The polar portion of the pharmaceutical composition prepared by the method according to any one of claims 1-8.

10. The use of the polar portion of the pharmaceutical composition prepared by any one of claims 1-8 in the preparation of anti-inflammatory, antipyretic, and analgesic drugs.

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