Pharmaceutical composition, preparation and preparation method

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition, a preparation and a preparation method. The pharmaceutical composition is prepared from the following components in parts by weight: 18.40 to 28.80 parts of baicalin, 0.216 to 0.336 part of forsythin, 1.12 to 1.92 parts of chlorogenic acid, 0.152 to 0.264 part of rutin, 0.944 to 1.608 parts of cryptochlorogenic acid, 0.568 to 1.176 parts of forsythiaside A, 0.192 to 0.468 part of isochlorogenic acid A, 0.152 to 0.336 part of isochlorogenic acid B, 0.376 to 0.924 part of isochlorogenic acid C, 0.32 to 0.756 part of oroxylin A glycoside, 0.936 to 1.44 part of neochlorogenic acid, 0.032 to 0.06 part of galuteolin and 0.544 to 1.20 part of forsythiaside I. The invention provides a traditional Chinese medicine active ingredient composition, and the active substance structure in the traditional Chinese medicine is complicated, so that the explanation of the medicine action mechanism and the material basis has great difficulty, which is the biggest obstacle influencing the trend to the world. According to the invention, the composition with better anti-inflammatory effect is developed by simplifying a split formula and combining specific active ingredients, and a foundation is also laid for internationalization of traditional Chinese medicines.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a pharmaceutical composition, a preparation and a preparation method. Background Art

[0002] In clinical treatment, Traditional Chinese Medicine (TCM) is widely used for respiratory diseases such as pneumonia, upper respiratory tract infections, and acute bronchitis. Commonly used single-herb herbs include heat-clearing and detoxifying drugs such as ephedra, gypsum, honeysuckle, forsythia, mint, ginger, and saposhnikovia divaricata, cough-relieving and expectorant drugs such as apricot kernel and platycodon, and Chinese patent medicines such as Ma Xing Shi Gan Tang and Wei Geng Tang, Yin Qiao San, and Ganmao Qingre Granules. The treatment of acute bronchitis focuses on relieving cough and asthma, commonly using herbs such as ephedra and apricot kernel, combined with prescriptions such as Xiao Qing Long Tang and Sang Ju Yin. Judging from the overall clinical results, it is safe, effective, and has few adverse reactions. Currently, the use of single-herb treatments has limited efficacy, and most commonly, multiple TCM herbs are used in combination.

[0003] However, each Chinese patent medicine contains many chemical components, and after extraction, there are also many ineffective components or components that have antagonistic effects on the active components. Due to the large number of components, the physiological effects of the drug are closely related to its complex chemical composition. Research on its active ingredients at home and abroad mainly focuses on medicinal materials and indications, and there is no in-depth study on the relationship between effective components and activity. In addition, the complex composition brings difficulties to the study of the material basis of drug action, which is an important reason hindering the internationalization of traditional Chinese medicine. Therefore, accurately simplifying the composition combination is of great significance for the explanation of the mechanism of drug action, and how to refine the active ingredients to achieve higher drug efficacy is a problem that needs to be solved urgently by those skilled in the art. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the present invention provides a pharmaceutical composition, a preparation and a preparation method.

[0005] In order to achieve the purpose of the present invention, the technical solutions adopted are as follows:

[0006] A pharmaceutical composition, comprising, by weight, 18.40-28.80 parts of baicalin, 0.216-0.336 parts of forsythiaside, 1.12-1.92 parts of chlorogenic acid, 0.152-0.264 parts of rutin, 0.944-1.608 parts of cryptochlorogenic acid, 0.568-1.176 parts of forsythiaside A, 0.192-0.468 parts of isochlorogenic acid A, 0.152-0.336 parts of isochlorogenic acid B, 0.376-0.924 parts of isochlorogenic acid C, 0.32-0.756 parts of melaleuca alternifolia A glycoside, 0.936-1.44 parts of neochlorogenic acid, 0.032-0.06 parts of luteolin, and 0.544-1.20 parts of forsythiaside I.

[0007] In some embodiments of the present invention, a composition having a good therapeutic effect on tonsillitis and pneumonia comprises, by weight, 20.70-25.30 parts of baicalin, 0.243-0.308 parts of forsythiaside, 1.26-1.76 parts of chlorogenic acid, 0.171-0.242 parts of rutin, 1.062-1.474 parts of cryptochlorogenic acid, 0.639-1.076 parts of forsythiaside A, 0.216-0.429 parts of isochlorogenic acid A, 0.171-0.308 parts of isochlorogenic acid B, and 1.26-1.76 parts of chlorogenic acid. 0.423-0.847 parts, melaleuca alternifolia A glycoside 0.36-0.693 parts, neochlorogenic acid 1.053-1.32 parts, luteolin 0.036-0.055 parts, forsythiaside I 0.612-1.10 parts.

[0008] In some embodiments of the present invention, in the composition having a good therapeutic effect on tonsillitis and pneumonia, the sum of the isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C is not higher than 1.4 parts by weight.

[0009] In some embodiments of the present invention, a composition having a good therapeutic effect on tonsillitis and pneumonia comprises, by weight, 23.0-28.80 parts of baicalin, 0.216-0.27 parts of forsythiaside, 1.12-1.40 parts of chlorogenic acid, 0.152-0.19 parts of rutin, 0.944-1.18 parts of cryptochlorogenic acid, 0.568-0.71 parts of forsythiaside A, 0.192-0.24 parts of isochlorogenic acid A, 0.152-0.19 parts of isochlorogenic acid B, 0.47-0.924 parts of isochlorogenic acid C, 0.32-0.63 parts of melaleuca alternifolia A glycoside, 0.936-1.20 parts of neochlorogenic acid, 0.032-0.047 parts of luteolin, and 0.544-0.68 parts of forsythiaside I.

[0010] Preferably, the pharmaceutical composition comprises, by weight: 23.0 parts of baicalin, 0.27 parts of forsythiaside, 1.40 parts of chlorogenic acid, 0.19 parts of rutin, 1.18 parts of cryptochlorogenic acid, 0.71 parts of forsythiaside A, 0.24 parts of isochlorogenic acid A, 0.19 parts of isochlorogenic acid B, 0.47 parts of isochlorogenic acid C, 0.63 parts of melaleuca alternifolia A glycoside, 1.20 parts of neochlorogenic acid, 0.047 parts of luteolin, and 0.68 parts of forsythiaside I.

[0011] The second object of the present invention is to provide a pharmaceutical preparation, the raw materials of which include the pharmaceutical composition.

[0012] Preferably, the pharmaceutical preparation further comprises pharmaceutically acceptable excipients.

[0013] Preferably, the dosage form of the pharmaceutical preparation is selected from oral preparations or injections, and the oral preparation is selected from mixtures, granules, capsules or tablets.

[0014] The third object of the present invention is to provide a use of the pharmaceutical composition in the preparation of medicines for treating pneumonia, upper respiratory tract infection or acute bronchitis.

[0015] Preferably, the pneumonia is community-acquired pneumonia, and the upper respiratory tract infection is tonsillitis.

[0016] In some embodiments of the present invention, the pharmaceutical composition or drug can be prepared into dosage forms such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, and lozenges. Suitable drug carriers in the art can be selected for different dosage forms.

[0017] The pharmaceutical carrier used can be a solid, liquid or gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0018] When preparing oral dosage compositions, any medium consistent with pharmaceutical formulation requirements can be used. For example, water, ethanol, oils, alcohols, flavoring agents, preservatives, colorants, and the like can be used to form oral liquid formulations, such as suspensions and solutions; while carriers, such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, emulsifiers, lubricants, binders, and disintegrants can be used to form oral solid formulations, such as powders, capsules, and tablets. Tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or nonaqueous techniques.

[0019] Tablets containing the Chinese medicine composition of the present invention can be prepared by tableting or molding, and one or more auxiliary ingredients or adjuvants can be used. The active ingredient can be compressed into a free-flowing form (such as powder or granules) in a suitable machine, and can be prepared by mixing with a binder, lubricant, inert diluent, surfactant or dispersant. Molded tablets can be molded in a suitable machine, i.e., a powdered mixture moistened with an inert liquid diluent, preferably containing about 0.05 mg to about 5 g of active ingredient per tablet, and preferably containing about 0.05 mg to about 5 g of active ingredient per sachet or capsule. For example, a preparation intended for oral administration to humans may contain about 0.5 mg to about 5 g of active drug, mixed with an appropriate and convenient pharmaceutical carrier, which may account for about 5% to 95% of the total composition. Unit dosage forms typically contain about 1 mg to about 2 g of active ingredient, typically 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

[0020] In addition to the above-mentioned carrier components, the above-mentioned pharmaceutical preparations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. In addition, other excipients, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, colorants and flavoring agents, etc., may be added, and the components containing the Chinese medicine composition of the present invention may also be prepared in the form of powder or concentrate.

[0021] In some embodiments, the pharmaceutical composition of the present invention is prepared in the form of an injection.

[0022] In some embodiments, the injection comprises a sterile solution.

[0023] In some embodiments, the present invention comprises at least one preservative. Examples of suitable preservatives that can be used include, but are not limited to, phenol, benzyl alcohol, m-cresol, methylparaben, propylparaben, butylparaben, chlorobutanol, thimerosal, phenylmercuric salts, and the like, or mixtures thereof.

[0024] The present invention comprises a pharmaceutically acceptable carrier, which comprises an aqueous carrier, such as water for injection. It may also comprise one or more pH adjusters, buffers, isotonic agents or chelating agents.

[0025] In one embodiment, the pH can be adjusted within the range by means of a pH adjusting agent such as sodium bicarbonate, lactic acid, hydrochloric acid, acetic acid, sodium hydroxide, etc. A buffer or buffer system can be used to maintain the pH within the desired range. Suitable buffers that can be used include, but are not limited to, lactate buffer, acetate buffer, citrate buffer, gluconate buffer, tartrate buffer, phosphate buffer, etc.

[0026] In a preferred embodiment, the osmotic pressure molarity can be adjusted with the help of an isotonic agent. Typical isotonic agents that can be used in sterile solutions include, but are not limited to, mannitol, sorbitol, sodium chloride, potassium chloride, lactose, sucrose, maltose, trehalose, dextrose, and mixtures thereof.

[0027] In a preferred embodiment, the sterile solution optionally contains a chelating agent, such as disodium EDTA, edetic acid, EDTA, and the like.

[0028] Terms and Claims of the Present Invention:

[0029] The articles "a," "an," and "the" include plural referents unless expressly limited to one or more referents otherwise.

[0030] Unless otherwise expressly indicated, all ranges or ratios disclosed herein are to be understood to include any and all subranges or subratios contained therein. For example, a range or ratio stated as 1 to 30 is to be considered inclusive of any and all subranges or subratios, integers, decimals, or subranges or subratios comprised therein, between a minimum of 1 and a maximum of 30, and including any subranges or subratios, integers, decimals, or subranges or subratios comprised therein.

[0031] The terms "comprises," "includes," "have," "has," "may," "contain" and variations thereof are meant as open-ended conjunctions or terms that do not exclude the possibility of additional compositions or structures.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] (1) The present invention combines the components to achieve a significantly better synergistic effect. The pharmaceutical composition can be prepared into an oral preparation or an injection, both of which can achieve more efficient anti-inflammatory effects and effectively improve the effects of treating tonsillitis and pneumonia.

[0034] (2) The present invention provides a composition of active ingredients in traditional Chinese medicine. The complex structure of active substances in traditional Chinese medicine makes it extremely difficult to explain its drug action mechanism and material basis, which is the biggest obstacle to its globalization. The present invention develops a composition with better anti-inflammatory effects by simplifying the prescription and combining specific active ingredients, laying the foundation for the internationalization of traditional Chinese medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 In the muscle irritation test, 72 hours after the end of drug administration, the quadriceps femoris muscles of the animals in the drug administration group were observed by eye. From left to right, they are the left quadriceps femoris and the right quadriceps femoris.

[0036] Figure 2 In the muscle irritation test, 72 hours after the end of drug administration, the quadriceps muscles of the control group were observed by eye. From left to right, they are the left quadriceps and the right quadriceps.

[0037] Figure 3 This is a pathological observation of the injected local muscle fibers in the drug-treated group 72 hours after the end of the drug administration in the muscle irritation test;

[0038] Figure 4 This is a pathological observation of the injected local muscle fibers in the control group 72 hours after the end of the drug administration in the muscle irritation test. DETAILED DESCRIPTION

[0039] The present invention will be further described below in conjunction with specific embodiments. The following raw materials are all commercially available conventional raw materials.

[0040] Example 1

[0041] This embodiment provides a pharmaceutical composition, which comprises, by weight, 23.0 parts of baicalin, 0.27 parts of forsythiaside, 1.40 parts of chlorogenic acid, 0.19 parts of rutin, 1.18 parts of cryptochlorogenic acid, 0.71 parts of forsythiaside A, 0.24 parts of isochlorogenic acid A, 0.19 parts of isochlorogenic acid B, 0.47 parts of isochlorogenic acid C, 0.63 parts of melaleucaside A, 1.20 parts of neochlorogenic acid, 0.047 parts of luteolin, and 0.68 parts of forsythiaside I.

[0042] The preparation method is as follows: the above components are mixed evenly.

[0043] Example 2

[0044] This embodiment provides a pharmaceutical composition, which comprises, by weight, 28.80 parts of baicalin, 0.216 parts of forsythiaside, 1.12 parts of chlorogenic acid, 0.152 parts of rutin, 0.944 parts of cryptochlorogenic acid, 0.568 parts of forsythiaside A, 0.192 parts of isochlorogenic acid A, 0.152 parts of isochlorogenic acid B, 0.924 parts of isochlorogenic acid C, 0.32 parts of melaleucaside A, 0.936 parts of neochlorogenic acid, 0.032 parts of luteolin, and 0.544 parts of forsythiaside I.

[0045] The preparation method is as follows: the above components are mixed evenly.

[0046] Example 3

[0047] This embodiment provides a pharmaceutical composition, which comprises, by weight, 18.40 parts of baicalin, 0.336 parts of forsythiaside, 1.92 parts of chlorogenic acid, 0.264 parts of rutin, 1.608 parts of cryptochlorogenic acid, 1.176 parts of forsythiaside A, 0.468 parts of isochlorogenic acid A, 0.336 parts of isochlorogenic acid B, 0.376 parts of isochlorogenic acid C, 0.756 parts of melaleucaside A, 1.44 parts of neochlorogenic acid, 0.06 parts of luteolin, and 1.20 parts of forsythiaside I.

[0048] The preparation method is as follows: the above components are mixed evenly.

[0049] Example 4

[0050] This embodiment provides a pharmaceutical composition, which comprises, by weight, 25.30 parts of baicalin, 0.243 parts of forsythiaside, 1.26 parts of chlorogenic acid, 0.171 parts of rutin, 1.062 parts of cryptochlorogenic acid, 0.639 parts of forsythiaside A, 0.216 parts of isochlorogenic acid A, 0.171 parts of isochlorogenic acid B, 0.847 parts of isochlorogenic acid C, 0.36 parts of melaleucaside A, 1.053 parts of neochlorogenic acid, 0.036 parts of luteolin, and 0.612 parts of forsythiaside I.

[0051] The preparation method is as follows: the above components are mixed evenly.

[0052] Example 5

[0053] This embodiment provides a pharmaceutical composition, which comprises, by weight, 20.70 parts of baicalin, 0.308 parts of forsythiaside, 1.76 parts of chlorogenic acid, 0.242 parts of rutin, 1.474 parts of cryptochlorogenic acid, 1.076 parts of forsythiaside A, 0.429 parts of isochlorogenic acid A, 0.308 parts of isochlorogenic acid B, 0.423 parts of isochlorogenic acid C, 0.693 parts of melaleucaside A, 1.32 parts of neochlorogenic acid, 0.055 parts of luteolin, and 1.10 parts of forsythiaside I.

[0054] The preparation method is as follows: the above components are mixed evenly.

[0055] Comparative Example 1

[0056] Compared with Example 1, the content of isochlorogenic acid B is higher, and the sum of isochlorogenic acid ABC exceeds 1.4 parts.

[0057] The proportions of the ingredients are as follows: 23.0 parts of baicalin, 0.27 parts of forsythiaside, 1.40 parts of chlorogenic acid, 0.19 parts of rutin, 1.18 parts of cryptochlorogenic acid, 0.71 parts of forsythiaside A, 0.39 parts of isochlorogenic acid A, 1.33 parts of isochlorogenic acid B, 0.77 parts of isochlorogenic acid C, 0.63 parts of melaleuca alternifolia A glycoside, 1.20 parts of neochlorogenic acid, 0.047 parts of luteolin, and 0.68 parts of forsythiaside I.

[0058] The preparation method is as follows: the above components are mixed evenly.

[0059] Comparative Example 2

[0060] The difference between this comparative example and Example 1 is that the components are different, forsythiaside I is replaced by forsythiaside B, and forsythiaside A is replaced by forsythiaside and forsythiaside A in a mass ratio of 1:1.

[0061] Comparative Example 3

[0062] The difference between this comparative example and Example 1 is that the components are different, and baicalin is replaced by wogonin and baicalin in a mass ratio of 1:1.

[0063] Effect experiment example:

[0064] Experimental Example 1: Effect on Tonsillitis

[0065] SD rats, weighing 180-220g, half male and half female, were randomly divided into 9 groups, each with 6 rats. They were respectively a blank group, a model group, a positive control group, Example 1-3 groups, and Comparative Example 1-3 groups. Except for the blank group, the rats in the other groups were sprayed with 15% ammonia water by mass twice a day using a medical laryngeal sprayer, once in the morning and once in the afternoon, with 3 sprays each time, for 3 consecutive days. Starting from the second day of modeling, the rats in each modeling group gradually developed itching, scratching the mouth and throat, increased oral secretions, significantly increased water intake, decreased food intake, and reduced autonomous activity. On the third day of modeling, the pharynx was congested and swollen, indicating that the modeling was successful.

[0066] Three days after modeling, rats were gavage-administered. Rats in the positive control group were given 3 mg / kg of dexamethasone. Rats in the Example and Comparative Example groups were given the compositions prepared in the corresponding groups at a dose of 120 mg / kg. The blank and model groups were given normal saline at a volume of 20 ml / kg for five consecutive days. 24 hours after the last dose, 1 mL of abdominal aortic blood was collected from each group of rats and placed in EDTA blood collection tubes. The number of white blood cells and lymphocytes in whole blood was determined using a TEK-VET5 blood cell analyzer.

[0067] Results: After 5 days of treatment, the redness and swelling of the pharynx of rats in all treatment groups improved compared with the model group. White blood cell counts and lymphocyte percentages in the model group were significantly higher than those in the blank group (P < 0.01). Compared with the model group, all treatment groups showed significant decreases in these indicators (P < 0.01). See Table 1 for details.

[0068] Table 1

[0069] Group <![CDATA[White blood cells (×10 9 cells / L)]]> Lymphocytes (%) Blank group 8.13±0.84 74.35±1.42 Model Group <![CDATA[18.25±1.62 ** ]]> <![CDATA[92.18±2.76 ** ]]> Positive control group <![CDATA[10.27±1.05 ## ]]> <![CDATA[75.66±1.63 ## ]]> Example 1 group <![CDATA[10.56±0.79 ## ]]> <![CDATA[76.01±1.74 ## ]]> Example 2 group <![CDATA[11.02±1.39 ## ]]> <![CDATA[78.22±2.09 ## ]]> Example 3 group <![CDATA[11.19±1.44 ## ]]> <![CDATA[78.48±2.35 ## ]]> Comparative Example 1 <![CDATA[11.34±1.67 ##& ]]> <![CDATA[80.10±2.48 ##& ]]> Comparative Example 2 <![CDATA[12.55±1.36 ##& ]]> <![CDATA[80.37±2.62 ##& ]]> Comparative Example 3 <![CDATA[12.68±1.59 ##& ]]> <![CDATA[81.14±2.51 ##& ]]>

[0070] Note: Compared with the blank group, ** P<0.01; compared with the model group, ## P<0.01. Compared with the Example 1 group, & P<0.05.

[0071] In some embodiments of the present invention, the compositions of Examples 4 and 5 also have therapeutic effects on tonsillitis and pneumonia comparable to those of Examples 1, 2, and 3.

[0072] Experimental Example 2: Experimental efficacy against pneumonia

[0073] Healthy male ICR mice, weighing 18±2 g, were adaptively fed for 5 days and lipopolysaccharide (LPS) was administered intranasally to establish the model. The mice were fasted but not watered for 12 h before modeling. After anesthesia with an intraperitoneal injection of 3% sodium pentobarbital solution, LPS solution (5 mg / kg, 2.5 mL / kg) was dripped into the nasal cavity. The mice were shaken up and down to ensure that the LPS solution was evenly distributed in the lungs.

[0074] Modeling mice were randomly divided into 8 groups of 10 mice each: a model group, a positive control group, Example 1-3 groups, and Comparative Example 1-3 groups. Ten normal mice were also selected as a blank group. The Example and Comparative Example groups received daily tail vein injections of the corresponding composition solution at a dose of 150 mg / kg (10 mL / kg). The positive control group received a tail vein injection of 50 mg / kg azithromycin, while the blank and model groups received an equal volume of 0.9% sodium chloride injection. Dosing was performed once daily for 5 consecutive days.

[0075] 24 hours after the last administration, the eyeballs of the mice were removed and blood was collected. The blood was centrifuged at 3000 rpm for 15 min, and the supernatant was used to measure the secretion levels of serum inflammatory factors (TNF-α, IL-6) by enzyme-linked immunosorbent assay.

[0076] Results: The levels of serum inflammatory factors (TNF-α and IL-6) in the model group were significantly higher than those in the blank group (P < 0.01), indicating successful modeling. Each treatment group significantly downregulated abnormally elevated serum inflammatory factors (TNF-α and IL-6) levels (P < 0.01), as shown in Table 2.

[0077] Table 2

[0078] Group IL-6 (pg / mL) TNF-α (pg / mL) Blank group 33.43±2.16 50.67±3.29 Model Group <![CDATA[95.85±5.94 ** ]]> <![CDATA[136.18±7.53 ** ]]> Positive control group <![CDATA[45.32±2.40 ## ]]> <![CDATA[58.31±3.75 ## ]]> Example 1 group <![CDATA[42.41±2.22 ## ]]> <![CDATA[56.97±3.62 ## ]]> Example 2 group <![CDATA[48.81±3.19 ## ]]> <![CDATA[59.93±4.37 ## ]]> Example 3 group <![CDATA[49.04±2.96 ## ]]> <![CDATA[60.84±4.82 ## ]]> Comparative Example 1 <![CDATA[51.92±3.64 ##& ]]> <![CDATA[66.70±5.10 ##& ]]> Comparative Example 2 <![CDATA[52.55±3.58 ##& ]]> <![CDATA[70.23±6.24 ##& ]]> Comparative Example 3 <![CDATA[54.17±4.01 ##& ]]> <![CDATA[71.45±6.65 ##& ]]>

[0079] Note: Compared with the blank group, ** P<0.01; compared with the model group, ## P<0.01. Compared with the Example 1 group, & P<0.05.

[0080] Experimental Example 3 Safety Experiment

[0081] 3.1 Acute toxicity

[0082] Kunming mice, weighing 18-20 g, half male and half female, were randomly divided into 6 groups of 10 mice each. Adaptive feeding was performed for 3 days. After fasting for 24 hours, the composition of Example 1 was injected into the tail vein once. The six groups of animals were administered with concentrations of 204.9 mg / mL, 174.1 mg / mL, 148.0 mg / mL, 125.8 mg / mL, 106.9 mg / mL, and 90.9 mg / mL, respectively, at a volume of 0.5 mL / 20 g body weight. After administration, the animals were observed for toxic reactions and mortality once daily for 7 consecutive days. The LD50 values ​​were calculated.

[0083] Another 60 mice were taken and operated according to the above method, except that the drug administered was the composition of Comparative Example 1.

[0084] As a result, the intravenous LD50 value and 95% confidence limit of the composition of Example 1 were 4954±579.6 mg / kg; the intravenous LD50 value and 95% confidence limit of the composition of Comparative Example 1 were 3411.46±323.13 mg / kg.

[0085] 3.2 Hemolysis and coagulation

[0086] Preparation of test sample: Dissolve the composition of Example 1 in 1 mL of water for injection, and then dilute to 50 mL with sodium chloride injection (concentration 12 mg / mL).

[0087] Preparation of a 2% red blood cell suspension: Collect approximately 10 mL of blood from the central ear artery of a rabbit. Place the blood in a beaded flask and shake for 10 minutes. Add 0.9% sodium chloride injection and shake thoroughly. Transfer the suspension into 10 mL graduated centrifuge tubes (8 mL per tube, for a total of 5 tubes). Centrifuge at 2500 rpm for 5 minutes and discard the supernatant. Add 6 mL of solvent to each tube and centrifuge again. Repeat the washing of the red blood cells 3-4 times until the supernatant is colorless and transparent. Add the resulting red blood cells to a 2% red blood cell suspension using 0.9% sodium chloride injection according to volume.

[0088] Experimental Method: Take seven clean test tubes and number them: tubes 1-5 are test sample tubes, tube 6 is the negative control tube, and tube 7 is the positive control tube. Add 2% red blood cell suspension, 0.9% sodium chloride solution, distilled water, and test sample solution in the order shown in Table 3.

[0089] Table 3

[0090] Test tube number 1 2 3 4 5 6 7 2% red blood cell suspension (mL) 2.5 2.5 2.5 2.5 2.5 2.5 2.5 0.9% sodium chloride solution (mL) 2.0 2.1 2.2 2.3 2.4 2.5 0 Distilled water (mL) 0.0 0.0 0.0 0.0 0.0 0.0 2.5 Test solution (mL) 0.5 0.4 0.3 0.2 0.1 0.0 0.0

[0091] After 3 hours, shake each tube and accurately pipette 0.1 mL. Dilute 50-fold with 0.9% sodium chloride injection. Shake well and add to the counting chamber of a counting chamber. Cover with a glass slide and let stand for a moment. Count the red blood cells under a microscope, counting the number of cells in the four corner squares and the central square, for a total of five squares. Repeat this sampling twice and calculate the average. Calculate the hemolysis degree of each tube according to the following formula.

[0092] Hemolysis degree (%) = (number of red blood cells in blank control tube - number of red blood cells in test tube) ÷ number of red blood cells in blank control tube × 100%

[0093] Hemolysis is considered to occur if the red blood cell hemolysis rate is greater than 30%.

[0094] According to the above method, the hemolysis of the composition of Comparative Example 1 was simultaneously detected.

[0095] result:

[0096] The test results of the composition of Example 1 are shown in Table 4 below.

[0097] Table 4

[0098]

[0099]

[0100] The test results of the composition of Comparative Example 1 are shown in Table 5 below.

[0101] Table 5

[0102]

[0103] The hemolysis rate of Example 1 was significantly lower than that of the comparative example 1, indicating better safety.

[0104] 3.3 Muscle irritation test

[0105] Test method: 12 male Japanese big-eared white rabbits, 7-8 months old, 2.27±0.18 kg. Divided into 2 groups, the drug-treated group and the control group, with 6 rabbits in each group. The left and right side self-comparison method was used. The 6 rabbits in the drug-treated group were injected with the solution of the composition of Example 1 into the quadriceps femoris on the left hind limb (before use, first fully dissolve it with 1 mL of sterile injection water, and then dilute it with sodium chloride injection to 100 mL (concentration 12 mg / mL)), and the right quadriceps femoris was injected with 0.9% sodium chloride injection, with a dosage volume of 0.5 mL / rabbit. The 6 rabbits in the control group were injected with the solution of the composition of Example 1 of the same concentration and volume into the quadriceps femoris on the left hind limb, and the right quadriceps femoris was injected with 0.9% sodium chloride injection. Before administration, the quadriceps femoris of the rabbit's hind limb was locally depilated with a baby push-up, and the depilated area was approximately 3×3 cm 2 After disinfection with 75% alcohol, fix the hind limb with the left hand and confirm the injection site. Hold the syringe filled with the medicine with the right hand and insert the needle into the muscle about 2.5 cm deep. After withdrawing the needle plug and no blood returns, inject the medicine.

[0106] 72 hours and 17 days after the last administration, three animals were anesthetized with 2% sodium pentobarbital (40 mg / kg) and sacrificed by carotid exsanguination. Muscles at the injection sites on both sides were harvested and observed for changes in the muscles with the naked eye and scored according to Table 6. The muscles were fixed with 4% formalin solution and sliced ​​for pathological observation.

[0107] Table 6

[0108] stimulus response Grading No significant changes 0 Mild congestion, or less than 0.5 × 1.0 cm 1 Moderate congestion, or an area larger than 0.5 × 1.0 cm 2 Severe congestion with muscle degeneration 3 Necrosis and brown degeneration 4 Extensive necrosis occurs 5

[0109] Observe with naked eyes, see picture - Figure 2 . Figure 1 In the muscle irritation test, 72 hours after the end of drug administration, the quadriceps femoris muscles of the animals in the drug administration group were observed by eye. From left to right, they are the left quadriceps femoris and the right quadriceps femoris.

[0110] Figure 2 In the muscle irritation test, 72 hours after dosing, the quadriceps femoris muscles of the control group were observed. From left to right, the left quadriceps femoris muscle is shown, followed by the right quadriceps femoris muscle. Results showed that 72 hours after dosing, no abnormal changes, such as congestion, hemorrhage, or necrosis, were observed in the quadriceps femoris muscles of either group. At the end of the recovery period, no abnormal changes, such as congestion, hemorrhage, or necrosis, were observed in the quadriceps muscles of the remaining animals in either group.

[0111] Pathological observation: see Figure 3-Figure 4 . Figure 3 This is a pathological observation of the injected local muscle fibers in the drug-treated group 72 hours after the end of the drug administration in the muscle irritation test; Figure 4 This is a pathological observation picture of the local injected muscle fibers of the control group 72 hours after the end of administration in the muscle irritation test. The results showed that 72 hours after the end of administration, there were no abnormalities in the local injected muscle fibers on both sides of the 3 animals in the administration group. There were no abnormalities in the local injected muscle fibers on the control side of 1 animal in the control group, while local muscle fiber degeneration and necrosis, connective tissue hyperplasia, and inflammatory cell infiltration were observed on the administration side. At the end of the recovery period, no abnormalities were observed in the local injected muscle fibers on both the control and administration sides of the two groups of animals. It can be seen that the composition of Comparative Example 1 has a mild reversible stimulating effect on the muscles. The composition of Example 1 has no stimulating effect and is safer to use.

[0112] The above detailed description is a specific description of one feasible embodiment of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not depart from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. A pharmaceutical composition, characterized in that In parts by weight, the pharmaceutical composition includes: 18.40-28.80 parts of baicalin, 0.216-0.336 parts of forsythiaside, 1.12-1.92 parts of chlorogenic acid, 0.152-0.264 parts of rutin, 0.944-1.608 parts of cryptochlorogenic acid, 0.568-1.176 parts of forsythiaside A, 0.192-0.468 parts of isochlorogenic acid A, 0.152-0.336 parts of isochlorogenic acid B, 0.376-0.924 parts of isochlorogenic acid C, 0.32-0.756 parts of melaleuca alternifolia A glycoside, 0.936-1.44 parts of neochlorogenic acid, 0.032-0.06 parts of luteolin, and 0.544-1.20 parts of forsythiaside I.

2. The pharmaceutical composition according to claim 1, characterized in that In parts by weight, the pharmaceutical composition includes: 20.70-25.30 parts of baicalin, 0.243-0.308 parts of forsythiaside, 1.26-1.76 parts of chlorogenic acid, 0.171-0.242 parts of rutin, 1.062-1.474 parts of cryptochlorogenic acid, 0.639-1.076 parts of forsythiaside A, 0.216-0.429 parts of isochlorogenic acid A, 0.171-0.308 parts of isochlorogenic acid B, 0.423-0.847 parts of isochlorogenic acid C, 0.36-0.693 parts of melaleuca alternifolia A glycoside, 1.053-1.32 parts of neochlorogenic acid, 0.036-0.055 parts of luteolin, and 0.612-1.10 parts of forsythiaside I.

3. The pharmaceutical composition according to any one of claims 1 to 2, characterized in that In parts by weight, the sum of the isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C is not higher than 1.4 parts.

4. The pharmaceutical composition according to claim 1, characterized in that In parts by weight, the pharmaceutical composition comprises: 23.0-28.80 parts of baicalin, 0.216-0.27 parts of forsythiaside, 1.12-1.40 parts of chlorogenic acid, 0.152-0.19 parts of rutin, 0.944-1.18 parts of cryptochlorogenic acid, 0.568-0.71 parts of forsythiaside A, 0.192-0.24 parts of isochlorogenic acid A, 0.152-0.19 parts of isochlorogenic acid B, 0.47-0.924 parts of isochlorogenic acid C, 0.32-0.63 parts of melaleuca alternifolia A glycoside, 0.936-1.20 parts of neochlorogenic acid, 0.032-0.047 parts of luteolin, and 0.544-0.68 parts of forsythiaside I.

5. The pharmaceutical composition according to any one of claims 1 to 2, characterized in that In parts by weight, the pharmaceutical composition includes: 23.0 parts of baicalin, 0.27 parts of forsythiaside, 1.40 parts of chlorogenic acid, 0.19 parts of rutin, 1.18 parts of cryptochlorogenic acid, 0.71 parts of forsythiaside A, 0.24 parts of isochlorogenic acid A, 0.19 parts of isochlorogenic acid B, 0.47 parts of isochlorogenic acid C, 0.63 parts of melaleuca alternifolia A glycoside, 1.20 parts of neochlorogenic acid, 0.047 parts of luteolin, and 0.68 parts of forsythiaside I.

6. A pharmaceutical preparation, characterized in that The raw materials of the pharmaceutical preparation include the pharmaceutical composition according to any one of claims 1 to 5.

7. The pharmaceutical preparation according to claim 6, characterized in that The pharmaceutical preparation further includes pharmaceutically acceptable excipients.

8. The pharmaceutical preparation according to claim 6, characterized in that The dosage form of the pharmaceutical preparation is selected from oral preparations or injections, and the oral preparation is selected from mixtures, granules, capsules or tablets.

9. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of medicaments for treating pneumonia, upper respiratory tract infection or acute bronchitis.

10. The use according to claim 9, characterized in that The pneumonia is community-acquired pneumonia, and the upper respiratory tract infection is tonsillitis.

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