CN110664888A

CN110664888A - Anti-inflammatory and antibacterial pharmaceutical composition, and preparation method and application thereof

Info

Publication number: CN110664888A
Application number: CN201910919673.XA
Authority: CN
Inventors: 向梅先; 苏汉文; 甘大莉; 师军凤
Original assignee: South Central University for Nationalities
Current assignee: South Central Minzu University
Priority date: 2019-09-26
Filing date: 2019-09-26
Publication date: 2020-01-10

Abstract

The invention provides an anti-inflammatory and antibacterial pharmaceutical composition, which comprises the following extracts of raw materials in parts by weight: 130-190 parts of pomegranate bark, 120-160 parts of scutellaria baicalensis, 100-140 parts of garden burnet and 100-140 parts of golden cypress. In addition, the invention also provides a preparation method of the anti-inflammatory and antibacterial pharmaceutical composition. The anti-inflammatory and antibacterial medicinal composition is a pure Chinese medicinal preparation, has better anti-inflammatory and antibacterial effects, and has the advantages of small dosage, convenient taking, wide application range, exact curative effect, safety, reliability, no relapse after healing, no drug dependence and no side effect, and can achieve the effect of treating both principal and secondary aspects of diseases.

Description

Anti-inflammatory and antibacterial pharmaceutical composition, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of traditional Chinese medicine preparations, and particularly relates to an anti-inflammatory antibacterial pharmaceutical composition, and a preparation method and application thereof.

Background

Inflammation is a common and multiple disease, which is a reaction of living tissues to the production of inflammatory factors, mainly for prevention, and vascular reaction is a main link of the inflammation process, and is clinically manifested as red, swollen, hot, pain and dysfunction, and accompanied with systemic reactions such as fever, increase of white blood cells, proliferation of mononuclear-phagocyte system and the like, and is a process mediated by inflammatory mediator molecules or cytokines. Many molecules of inflammatory mediators are derivatives of arachidonic acid, and mainly include Prostaglandins (PGs), Leukotrienes (LTs), thromboxanes (thromboxanes), and the like. Any factor that can cause tissue damage can cause inflammation, i.e., inflammatory factors, which can be divided into five groups, among which biological factors: such as viruses, bacteria, fungi, etc.; physical factors: high temperature, low temperature, radioactive substances, ultraviolet rays, mechanical damage, and the like; chemical factors: such as strong acids, strong bases, etc.; foreign matter: foreign substances entering the human body through various routes, such as iron metal, debris, dust, etc.; allergic reactions, such as excessive immune reactions caused by abnormal immune response, can cause inflammation.

A series of medical studies have shown that inflammation is associated with stroke, migraine, cancer, osteoarthritis, asthma, senile dementia, arteriosclerosis, heart disease, periodontitis, chronic fatigue syndrome, etc., and thus, attention is paid to the risk of inflammation.

At present, the clinical anti-inflammatory drugs can be roughly divided into three categories, namely non-steroidal anti-inflammatory drugs (NSAIDs), steroidal anti-inflammatory drugs and traditional Chinese medicine anti-inflammatory drugs. The non-steroidal drugs mainly comprise: aspirin, acetaminophen, diclofenac, indomethacin (indomethacin), ibuprofen and the like, has weak non-steroidal anti-inflammatory effect, and has side effects of gastrointestinal tract reaction, liver toxicity, nervous system toxicity, cardiovascular disease risk and the like; the steroid anti-inflammatory drug is glucocorticoid, and general hormone drugs have strong anti-inflammatory action, but have side effects of osteoporosis, infection diffusion, central obesity, water and sodium retention and the like. Although the two drugs are applied clinically, the two drugs have strong side effects, and long-term and repeated application of the anti-inflammatory drugs can also negatively affect the immune function of the organism, aggravate the decline of the physique of patients caused by the prolonged course of disease and enable the vicious circle of the disease.

Therefore, the search for a substitute with good antibacterial and anti-inflammatory effects and small side effects from the Chinese medical treasury is great tendency. The antibacterial and anti-inflammatory effects of the traditional Chinese medicine are mostly derived from natural animal and plant active ingredients, and the compound compatibility of the traditional Chinese medicine can generate synergistic effect, enhance the immunity of patients and have better curative effect on infectious diseases, inflammations and the like. Therefore, the traditional Chinese medicine has the comprehensive advantage that the chemical medicine can not replace the traditional Chinese medicine for treating the inflammation, and has attracted wide attention in the medical field. The action mechanism of the traditional Chinese medicine for resisting inflammation is to play a role in resisting inflammation by regulating the function of affecting the hypothalamus-pituitary-adrenal (HPA) axis and inhibiting the release of inflammatory mediators. In recent years, research also finds that the traditional Chinese medicine can inhibit free radical substances such as Nitric Oxide (NO) and cytokines related to inflammation. Therefore, the anti-inflammatory activity of the traditional Chinese medicine draws wide attention at home and abroad.

Disclosure of Invention

The invention aims to overcome the problems that the existing steroidal and non-steroidal anti-inflammatory drugs have strong side effects, and the anti-inflammatory drugs can also negatively influence the immune function of the organism after being repeatedly applied for a long time, so that the physical deterioration of patients caused by the prolonged course of disease is aggravated, and the vicious circle of the disease is caused.

Therefore, the invention provides an anti-inflammatory and antibacterial pharmaceutical composition, which comprises the following extracts of raw materials in parts by weight: 130-190 parts of pomegranate bark, 120-160 parts of scutellaria baicalensis, 100-140 parts of garden burnet and 100-140 parts of golden cypress.

In a preferred embodiment, the medicinal composition comprises the following extracts of the raw materials in parts by weight: 150-170 parts of pomegranate bark, 130-150 parts of scutellaria baicalensis, 110-130 parts of garden burnet and 110-130 parts of golden cypress.

Further preferably, the effective ingredients comprise the following extracts of raw materials in parts by weight: 160 parts of pomegranate bark, 140 parts of scutellaria baicalensis, 120 parts of garden burnet and 120 parts of golden cypress.

The invention also provides a preparation method of the anti-inflammatory antibacterial pharmaceutical composition, which comprises the following steps:

1) weighing the raw material components according to the designed weight parts;

2) adding 60% ethanol into pericarpium Granati, soaking for 0.5h, heating and reflux extracting for 2 times, and filtering to obtain pericarpium Granati extractive solution;

3) extracting Scutellariae radix, radix Sangusorbae, and cortex Phellodendri with 60% ethanol for 2 times, and filtering to obtain extractive solution;

4) mixing the pomegranate bark extract in the step 2) and the Chinese medicinal material extract in the step 3) to obtain a mixed extract, and concentrating and drying the mixed extract to obtain the anti-inflammatory and antibacterial medicinal composition.

Further, the heating reflux extraction mode in the step 2) is as follows: extracting with 6 times of 60% ethanol for 15min, extracting with 4 times of 60% ethanol for 10min, and mixing the filtrates.

Further, the two extraction modes of the scutellaria baicalensis, the sanguisorba officinalis and the golden cypress in the step 3) are as follows: adding 60% ethanol 6 times the total weight of Scutellariae radix, radix Sangusorbae and cortex Phellodendri, extracting for 1 hr, filtering, adding 60% ethanol 4 times the total weight of Scutellariae radix, radix Sangusorbae and cortex Phellodendri into the residue, extracting for 0.5 hr, filtering, and mixing the filtrates.

Further, the mixed extracting solution in the step 4) is concentrated to a relative density of 1.25, and then spray drying is carried out.

Further, the method also comprises a step 5), adding auxiliary materials of crospovidone and sodium carboxymethyl starch as disintegrating agents and microcrystalline cellulose as filling agents into the concentrated and dried medicinal composition obtained in the step 4), uniformly mixing, preparing soft materials by using ethanol solution with volume concentration of 60%, granulating, drying at 50 ℃ to obtain granules, controlling air relative humidity below 66%, and pressing into dispersible tablets.

Further, the mass ratio of the pharmaceutical composition to the auxiliary materials is 100-20: 254-350, and the mass ratio of the crospovidone, the sodium carboxymethyl starch and the microcrystalline cellulose in the auxiliary materials is 1:1: 1.

The invention also provides application of the anti-inflammatory and antibacterial pharmaceutical composition in preparing medicaments for treating inflammation, wherein the inflammation comprises bacillary dysentery, upper respiratory tract infection, tonsillitis, gastritis and urinary tract infection.

In the present invention, the activity of Scutellaria baicalensis GeorgiThe component can inhibit inflammation molecule PGE₂And leukotriene C4 and nitric oxide NO synthesis and expression of certain adhesion molecules (such as ICAM-1), block the arachidonic acid pathway from different links, and can inhibit secretion release of cytokines (mainly including IL-12, IL-8, TNF-alpha, IL-6) and inhibit transcription activity of nuclear factor NF-KB; in addition, the total flavone of Scutellariae radix can inhibit COX-2 synthesis, thereby inhibiting inflammatory reaction and achieving anti-inflammatory effect. The sanguisorba has the effects of resisting bacteria, diminishing inflammation and enhancing immunity, and the antibacterial effect is mainly realized by destroying the integrity of cells and changing the permeability of cell membranes; the expression of Toll-like receptor 4 and IL-23 in intestinal mucosa is reduced, TNF-alpha is inhibited, IL-1 beta level is reduced, IL-10 content is increased, NF-kB protein activity is reduced, and the like, so that the anti-inflammatory effect is achieved, and the sanguisorba can also enhance the immunity. The active component tannin of pericarpium Granati has wide antibacterial spectrum and strong antibacterial activity, and the pericarpium Granati also has antiinflammatory activity, and its water extract can inhibit NO secretion induced by LPS in macrophage inflammatory reaction. The phellodendron has stronger inhibition effect on respiratory bacteria and RNA (ribonucleic acid) combination, and the mechanism of the phellodendron is related to the activity of inhibiting staphylococcus aureus, proteus and streptococcus A and B; in addition, cortex Phellodendri also has immunoregulatory effect, thereby relieving inflammation injury; has antiulcer and antioxidant effects, and can be used for inhibiting gastric ulcer. The above medicines have synergistic effect, and can achieve better effects of resisting bacteria, diminishing inflammation, resisting ulcer and enhancing immunity.

Compared with the prior art, the invention has the beneficial effects that:

the anti-inflammatory and antibacterial pharmaceutical composition provided by the invention is a pure traditional Chinese medicine preparation, has good anti-inflammatory and antibacterial effects, is small in dosage, convenient to take, wide in application range, exact in curative effect, safe and reliable, free of relapse after healing, free of drug property and side effects, and can achieve the effect of treating both principal and secondary aspects of diseases.

The present invention will be described in further detail below with reference to the accompanying drawings.

Drawings

FIG. 1 is a bar graph of the effect of different test group drugs on the mouse peritoneal capillary permeability in accordance with the present invention;

FIG. 2 is a graph showing the effect of the toe swelling experiment caused by egg white in the present invention;

FIG. 3 is a bar graph showing the rate of toe swelling caused by egg white for different test groups of drugs in the present invention;

FIG. 4 is a graph showing the effect of formaldehyde on toe swelling;

FIG. 5 is a bar graph of the change of toe swelling caused by formaldehyde in different test groups of drugs tested at different time periods in the present invention;

FIG. 6 is a bar graph of the effect of different test group drugs in the present invention on swelling of mouse pinna;

FIG. 7 is a graph showing the effect of dispersible tablets of the pharmaceutical composition of the present invention on gastric tissues of rats in gastric ulcer model;

FIG. 8 is a graph showing the results of HE staining of gastric tissues of rats in a gastric ulcer model with dispersible tablets of the pharmaceutical composition of the present invention;

FIG. 9 is a graph showing the result of PAS staining of gastric ulcer model rat stomach tissue by the dispersible tablet of the present invention;

FIG. 10 is a graph showing the results of HE staining of mice with dispersible tablets of the pharmaceutical composition of the present invention for acute lung injury;

FIG. 11 is a graph showing the effect of the pharmaceutical composition of the present invention on the body weight of rats;

FIG. 12 is a graph showing the effect of the pharmaceutical composition of the present invention on the feed intake of rats.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 160 parts of pomegranate bark, 140 parts of scutellaria baicalensis, 120 parts of garden burnet and 120 parts of golden cypress.

The preparation method of the anti-inflammatory and antibacterial pharmaceutical composition comprises the following steps:

(1) pulverizing pericarpium Granati into coarse powder, weighing pericarpium Granati coarse powder according to above formula, adding 6 times of 60% ethanol, heating and reflux extracting for 15min, filtering to obtain filtrate, adding 4 times of 60% ethanol into residue, extracting for 10min, filtering to obtain filtrate, and mixing the filtrates.

(2) Weighing the scutellaria baicalensis, the garden burnet and the golden cypress according to the prescription amount, crushing into coarse powder, adding 60% ethanol which is 6 times of the coarse powder for the first time, extracting for 1 hour, and filtering to obtain filtrate for later use; extracting the residue with 60% ethanol 4 times the coarse powder for 0.5 hr, filtering to obtain filtrate, mixing the filtrates

(3) And (3) combining the filtrates in the steps (1) and (2), recovering ethanol, and performing spray drying to obtain dry extract powder for later use.

(4) And (3) forming of the preparation: adding microcrystalline cellulose (MCC) serving as a filler into the dry extract powder obtained in the step (3), adding crospovidone (PVPP) and sodium carboxymethyl starch (CMS-Na) serving as disintegrants, adding the extract powder and auxiliary materials (PVPP: CMS-Na: MCC: 1:1:1) at a ratio of 1:2.54, granulating with 60% ethanol, drying at 50 ℃, and tabletting with the air relative humidity controlled below 65% to obtain the anti-inflammatory and antibacterial medicinal composition dispersible tablet.

Example 2:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 130 parts of pomegranate bark, 120 parts of scutellaria baicalensis, 100 parts of garden burnet and 100 parts of golden cypress.

The preparation method of the anti-inflammatory and antibacterial pharmaceutical composition of this example is the same as that of example 1.

Example 3:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 140 parts of pomegranate bark, 130 parts of scutellaria baicalensis, 110 parts of garden burnet and 110 parts of golden cypress.

Example 4:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 150 parts of pomegranate bark, 140 parts of scutellaria baicalensis, 120 parts of garden burnet and 120 parts of golden cypress.

Example 5:

Example 6:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 170 parts of pomegranate bark, 140 parts of scutellaria baicalensis, 120 parts of garden burnet and 120 parts of golden cypress.

Example 7:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 180 parts of pomegranate bark, 150 parts of scutellaria baicalensis, 130 parts of garden burnet and 130 parts of golden cypress.

Example 8:

the embodiment provides an anti-inflammatory and antibacterial pharmaceutical composition, the active ingredients of which comprise the following extracts of raw materials in parts by weight: 190 parts of pomegranate bark, 160 parts of scutellaria baicalensis, 140 parts of garden burnet and 140 parts of golden cypress.

The anti-inflammatory and antibacterial pharmaceutical composition prepared in example 1 was used as an example, and the antibacterial and anti-inflammatory effects thereof were studied by animal experiments and in vitro bacteriostatic tests.

1 dose

SD rat: the low, medium and high doses of the dispersible tablet of the pharmaceutical composition in this example are 0.25, 0.500 and 1.000g/kg respectively; sanjiuweitai (gastritis positive drug): 0.45 g/kg; globeflower soft capsule group (inflammation positive medicine): 0.30 g/kg.

KM mice: the low, medium and high doses of the pharmaceutical composition are respectively 0.312, 0.624 and 1.248 g/kg; the trollflower soft capsule group: 0.43 g/kg.

2 method of experiment

2.1 experiment on increase in Permeability of capillary vessels in the peritoneal Cavity of mice

The 50 mice were randomly divided into 5 groups, i.e.: normal group, positive group (the dosage of the trollius chinensis bunge soft capsule is 0.43g/kg), and dispersible tablets (the dosage is 0.312, 0.624 and 1.248g/kg) of the pharmaceutical composition, wherein each group contains 10 tablets. Gavage is continued for one week, once daily, 1h after the last administration, and tail vein injection of 0.5% Ivensan (dose of 0.1mL/10g) and intraperitoneal injection of 0.6% glacial acetic acid (0.2 mL/tube). The mice were sacrificed 30min later, 6mL of normal saline was intraperitoneally injected, the abdomen of the mice was gently kneaded, the saline rinse solution was collected, centrifuged at 1000r/min for 10min, and the OD value was measured at 590nm from the supernatant.

2.2 toe swelling due to egg white in rats

50 rats were randomly divided into 5 groups, i.e.: the model group, the trollius chinensis bunge soft capsule group (the dose is 0.30g/kg) and the pharmaceutical composition dispersible tablets (the dose is 0.25, 0.500 and 1.000g/kg), wherein 10 of the groups are continuously administrated for 7 days, after the last administration for 1 hour, 10% egg white (0.5mL/kg) is injected to the sole of the left hind paw of each rat, and the toe swelling volumes of the rats are detected by using a toe swelling determinator for 0, 0.5, 1, 2, 3, 4, 5 and 6 hours.

2.3 Effect of Formaldehyde on toe swelling in rats

50 rats were randomly divided into 5 groups, i.e.: model group, dispersible tablet of pharmaceutical composition (dosage 0.250, 0.500, 1.000g/kg), and positive group (trollius chinensis bunge soft capsule 0.30 g/kg). The administration was continued for 7 days. After 1h of the last administration, the newly prepared 2% formaldehyde solution was injected subcutaneously under the left hind paw of the rat (0.5mL/kg) and the volume of hind paw swelling was measured for 0, 5, 1, 2, 4, 6, 12, 24, 36, 72h using a toe swelling meter. The results were calculated according to the following formula:

the toe swelling rate is (volume of postinflammatory toe-volume of pro-inflammatory toe)/volume of pro-inflammatory toe.

2.4 mouse auricular swelling test

The 50 mice were randomly divided into 5 groups, i.e.: normal group, dispersible tablet of medicinal composition (dosage is 0.312, 0.624, 1.248g/kg), and positive group (trollius chinensis bunge soft capsule 0.43 g/kg). Continuously performing intragastric administration for 7 days, after the last administration for 1h, uniformly coating 20 mu L of dimethylbenzene on auricles of right ears of mice to cause inflammation, killing the mice after 30min of inflammation, and punching round ear plates at symmetrical positions of two ears respectively by using a punching machine to weigh.

Swelling degree is the right ear weight-left ear weight.

2.5 acute gastric ulcer in rats caused by ethanol

60 rats were randomly divided into 6 groups, i.e.: blank group, model group, medicinal composition dispersible tablet (0.25, 0.50, 1.00g/kg), positive group (Sanjiu stomach tai 0.45g/kg), the administration is performed by intragastric administration once a day, the administration is continued for 7 days, fasting is not prohibited from the 6 th day, after the last administration for 1h, the rest groups except physiological saline are administered by intragastric absolute ethyl alcohol 5mL/kg, the patients are killed after about 2h of modeling, the gastric cardia and pylorus are ligated by fine needle, the stomach is taken out after being cut off, the stomach tissue is taken for observation, then the stomach tissue is frozen and sliced, HE staining and PAS staining are performed, and the inflammatory pathological change condition of the stomach tissue is observed.

2.6 Effect of acute Lung injury in mice

60 mice were randomly divided into 6 groups, namely a blank group, a model group, a positive group (the dosage of the trollius chinensis bunge soft capsule is 0.43g/kg) and a medicinal composition dispersible tablet (the dosage is 0.25, 0.50 and 1.00g/kg), and 10 mice in each group. Continuously performing intragastric administration for one week, once daily, fasting for 12h before final administration without water prohibition, and after administration for 1h, performing combined induction by injecting LPS (1mg/ml, total injection amount is kept at 3mg/kg) into abdominal cavity of all rats except blank group and dropping 20 μ L of LPS with the same concentration into nose to establish acute lung injury model of mouseAnd (4) molding. After 12h of LPS induction, the mice were anesthetized with chloral hydrate and the abdominal aorta was bled. Standing at room temperature for 4 hr, collecting supernatant at 4 deg.C 3000r/30min, subpackaging at-80 deg.C, and treating PGE in serum with corresponding ELISA kit₂And detecting the content level of IL-1 beta, wherein the operation steps are carried out strictly according to the instruction of the kit. Lung tissue was taken and a portion was fixed with 10% neutral formaldehyde for HE staining.

2.7 dilution of dispersible tablet of pharmaceutical composition in vitro bacteriostasis test

Culturing and separating the obtained pathogenic bacteria, adding the aqueous solution of the dispersible tablet of the pharmaceutical composition respectively to perform in vitro antibacterial test, observing in vitro antibacterial effect, and screening out sensitive pathogenic bacteria.

3. Results and analysis

3.1 experiment on increase in Permeability of capillary vessels in the peritoneal Cavity of mice

The experimental result of the permeability of the capillary vessels in the abdominal cavity of the mouse shows that: compared with the control group, the dispersible tablet of the pharmaceutical composition of this example significantly inhibits the capillary permeability of the mouse abdominal cavity (P <0.01) in the high dose group, and the inhibition effects are similar, respectively 49% and 45.5%, and the results are shown in table 1 and fig. 1.

Table 1: effect of the dispersible tablets of the pharmaceutical composition on the capillary permeability of the mouse peritoneal cavity (n-10,

)

in comparison with the normal group,^*P<0.05，^**P<0.01。

3.2 toe swelling due to egg white in rats

The experimental results of the toe swelling of the rat caused by the egg white show that: the swelling of hind paw of the rats after molding is shown in fig. 2, and the toe swelling rate results are shown in table 2 and fig. 3. The low and medium doses (the doses are respectively 0.250g/kg and 0.500g/kg) of the dispersible tablet of the pharmaceutical composition can obviously inhibit toe swelling caused by egg white (P < 0.05); the high dose (dose is 1.000g/kg) can remarkably inhibit the hind paw swelling (P <0.01), which indicates that the dispersible tablet of the pharmaceutical composition has good anti-inflammatory effect.

Table 2: the medicine composition dispersing tablet can treat the toe swelling rate change caused by egg white (n is 10,

)

3.3 Effect on Formaldehyde-induced rat foot swelling

The results of the formaldehyde-induced toe swelling experiment of rats show that: as shown in FIG. 4, the hind paw of the rat was swollen after molding, and the molded part of the untreated hind paw was suppurative. The dispersible tablet of the pharmaceutical composition can obviously inhibit the toe swelling caused by formaldehyde at each administration dose (P <0.05 and P < 0.01); on the third day, the administration group had already substantially subsided swelling and had no suppuration phenomenon, while the positive control group was still red and swollen and had significant suppuration, indicating that the dispersible tablet of the pharmaceutical composition of the present invention has a good anti-inflammatory effect and can suppress the phenomenon of hind paw suppuration caused by formaldehyde, with the results shown in table 3 and fig. 5.

Table 3: the medicine composition dispersing tablet can treat the toe swelling rate change caused by formaldehyde (n is 10,

)

3.4 mouse ear swelling test

From the results of the mouse auricular swelling experiment, it can be seen that: compared with the normal group, the anti-inflammatory and antibacterial dispersible tablets have dose dependence on the inhibition of auricle swelling, and have obvious high-dose inhibition effect (^**P<0.01), the results are shown in table 4 and fig. 6.

Table 4: influence of anti-inflammatory and antibacterial dispersible tablet on mouse auricle swelling

3.5 acute gastric ulcer in rats caused by ethanol

The results of the ethanol induced acute gastric ulcer test in rats are shown in fig. 7, 8 and 9. The results show that the normal group SD rats have complete gastric mucosa structure without congestion, the arrangement rule of the stomach fovea is clear and visible, the gland is not damaged, bleeding and inflammatory cell infiltration are not seen in the lamina propria, and PAS staining shows that the mucosa is complete and has no shedding phenomenon. Compared with the blank control group, the rats in the model group have serious gastric mucosa injury, obvious congestion, submucosal edema, epithelial cell loss and inflammatory cell infiltration can be seen, and mucosa shedding is obvious. The administration group (i.e. the dispersible tablet of the pharmaceutical composition) then starts to recover: the symptoms of congestion, mucosa exfoliation, epithelial cell loss, inflammatory cell infiltration and the like are relieved, and the administration group of the dispersible tablet of the pharmaceutical composition is dose-dependent, the dose is increased, and the damage is lighter.

3.6 pathological Effect on LPS-induced acute Lung injury mice

The lung tissue experimental results of the mice with acute lung injury are shown in fig. 10, the lung tissues in the normal group are uniformly arranged, the alveolar structure is clear and complete, the lung tracheal structure is obvious, and no obvious inflammatory cell infiltration and no red blood cell scattering exist. In the model group, the lung tissue arrangement is disordered, the alveolar structure is incomplete, the inflammatory infiltration is serious, and the erythrocyte infiltration is obvious. Compared with the model group, the positive group has complete alveolar structure, the inflammatory infiltration is obviously reduced, and a small amount of red blood cell infiltration exists; in the medicine composition dispersing tablet group, the inflammatory infiltration is obviously relieved along with the increase of the administration dosage, the structural integrity of the alveolus is increased, and the dispersion of red blood cells is gradually reduced.

3.7 in vitro bacteriostasis results of series dilutions of dispersible tablets of pharmaceutical composition

The results of the in vitro antibacterial tests on dispersible tablets of the drug combination are shown in table 5. The test result shows that: the dispersible tablet of the pharmaceutical composition has different degrees of bacteriostasis on tested strains (mainly common pathogenic bacteria and conditional pathogenic bacteria causing gastrointestinal and respiratory tract infection), has strong inhibition on escherichia coli and streptococcus pneumoniae, has consistent clinical treatment efficacy with the MIC of each bacterium being 128-32, and has strong antibacterial effect on most of tested strains.

Table 5: in vitro antibacterial experimental result of dispersible tablet of drug composition

Note: -sterile growth, + sterile growth

The experimental result shows that the inflammation is divided into infectious inflammation and non-infectious inflammation according to the infection way, the infectious inflammation is mainly caused by biological pathogens, the non-infectious inflammation comprises aseptic inflammation (non-specific inflammation) and allergic reaction inflammation, and the in-vitro bacteriostasis experiment shows that the pharmaceutical composition has antibacterial action with different degrees on tested strains, the MIC (minimal inhibitory concentration) on each strain is between 128 and 32, and the antibacterial action on most tested strains is strong; the mouse auricle swelling model is one of non-specific inflammation models, mainly takes chemical substances as a inflammation-causing agent to induce the mouse auricle swelling, and in recent years, the mouse auricle swelling model has developed into a mature animal inflammation model. The common inflammation-causing agents for mouse auricle swelling include xylene, arachidonic acid, etc. According to the invention, a classical model of xylene-induced mouse auricle swelling is adopted to study the anti-inflammatory activity of the pharmaceutical composition of the embodiment, and experimental results show that the pharmaceutical composition of the embodiment has good anti-inflammatory activity, and specifically can obviously inhibit mouse auricle swelling caused by xylene, and the inhibition rate of the pharmaceutical composition reaches 68.4%, which is better than that of a positive control. Aiming at the fact that the medicine can be clinically used for treating acute gastritis and enteritis, the research on the acute gastritis is carried out, and the result shows that compared with a control group, the gastric mucosa of a model group rat is seriously injured, obvious congestion, submucosa edema, epithelial cell loss and inflammatory cell infiltration can be seen, and the mucosa is obviously exfoliated. After the anti-inflammatory and antibacterial dispersible tablet is administered, the recovery starts, specifically, the symptoms such as congestion, mucosa shedding, epithelial cell loss and inflammatory cell infiltration are relieved, the dosage is dependent, the dosage is increased, the injury is lighter, and the animal experiment verifies the efficacy of the anti-inflammatory and antibacterial dispersible tablet for treating gastritis.

In order to observe the safety of the pharmaceutical composition of the present invention, the following examples of the anti-inflammatory and antibacterial pharmaceutical composition prepared in example 1 were conducted to study acute and long-term toxicity.

1 experimental part

1.1 reagent and animal

The target traditional Chinese medicine composition extract 1mL contains 9g of crude drug, and is diluted to the required concentration by distilled water.

Acute toxicity test: taking 40 healthy Kunming mice; the weight of the animal is 18-22 g (research center for laboratory animals in Hubei province) SCXK (Hubei) 2015-0018, and the license number of the center for the laboratory animals: 2016-0089.

Long-term toxicity test: SD rat, weight 80 ~ 120g (Hubei province laboratory animal research center) license number: SCXK (jaw) 2015-0018.

1.2 methods and results

1.2.1 oral maximum tolerated dose determination: taking 40 healthy Kunming mice with the weight of 18-22 g, and randomly dividing the mice into a blank control group and an administration group according to the weight, wherein each group comprises 20 mice with half of male and female. Fasting is not prohibited for 12 hours before gavage, the administration concentration is 0.25g/mL, gavage administration is carried out for 2 times according to the administration volume of 40mL/kg at intervals of 8 hours, a blank control group is administered with physiological saline with the same volume, the reaction of animals is observed after the administration, the animal behaviors, the appearance, the food intake, the excrement, the size and the pathological change of main organs and the death condition are observed and recorded for 14 days continuously, and the maximum administration amount of the mice to the dispersible tablet of the pharmaceutical composition is calculated to be 20 g/kg.

1.2.2 Long term toxicity test: selecting 80 rats with half male and female, and dividing each group into a blank group and three dosage groups of low, medium and high (1.25g/kg, 2.5g/kg, 5g/kg) according to a weight balance method, wherein each group comprises 20 rats; a control group was also provided and an equal volume of physiological saline was administered. After adaptive feeding for one week, performing intragastric administration at a ratio of 2.0mL/100g, administering once a day for 6 days per week for 4 weeks, stopping administration after 4 weeks, and recovering for a period of timeOnce a week, weigh once a week and adjust the dosage. The general conditions (activity, hair color, food intake, drinking and stool) of the rats were closely observed during the administration period. After 12 weeks, blood was collected, blood routine and biochemical index measurements were performed using XN9000 model blood analyzer and AU-2400 model biochemical analyzer, and then rats were sacrificed and vital organs were immediately weighed for wet weight, organ coefficients were calculated and histology was performed. Data to

Statistical analysis was performed using the inter-group t-test.

The general conditions are that rats in each dose group have normal activity, smooth hair color, no abnormality in food intake, water drinking and urine, continuous weight increase, occasional defecation and no significant difference among groups; the results of the effects of the pharmaceutical compositions on the body weight of male and female rats are shown in tables 6, 7 and FIG. 11, and the effects of the pharmaceutical compositions on the food intake of rats are shown in FIG. 12.

Peripheral blood routine, rats were not abnormal in each hemogram index for 4 weeks, and no significant difference (P >0.05) was observed between each group and the control group, and the results are shown in Table 8.

Biochemical index after 4 weeks, the biochemical indexes of the blood of the rats are in a normal range, and no significant difference (P >0.05) is generated between each group and a control group, and the results are shown in Table 9.

The results of histomorphometry of rat pathological tissues at the end stage of experiment and the recovery stage show that the control group of animals has no abnormal histological change in the tissues of the organs, and the high-dose group of animals has no abnormal histological change caused by drugs, and has no significant difference (P is more than 0.05) compared with the control group, and the results are shown in tables 10 and 11.

Table 6: effect of pharmaceutical composition on body weight of Male rat

Table 7: effect of pharmaceutical composition on body weight of female rat

Table 8: general Effect of pharmaceutical composition on peripheral blood of rat

Table 9: biochemical influence of pharmaceutical composition on peripheral blood of rat

Table 10: influence of the pharmaceutical composition on organ coefficients of male rat

Table 11: influence of the pharmaceutical composition on organ coefficients of male rat

The toxicity test studies described above show that: the extract of the traditional Chinese medicine composition is used for intragastric administration, the administration dosage is 20 times of the dosage of adults, no animal death still exists, and the traditional Chinese medicine composition has no obvious oral toxicity; and the general condition, weight gain, organ coefficient, hemogram and blood biochemical indexes of the rat after being taken for 4 weeks have no obvious difference compared with a blank control group, and histological examination proves that other important organs except the lung have no abnormal change, which indicates that the dispersible tablet of the pharmaceutical composition is safe for long-term use.

The pharmaceutical compositions of the above examples 2-8 were also tested for antibacterial, anti-inflammatory and anti-gastritis using animal experiments and in vitro bacteriostatic tests, and the results were the same as those of example 1, all of which had better antibacterial and anti-inflammatory activities, no significant oral toxicity, and could be taken for a long period of time.

The clinical effect is as follows:

case 1:

21 gastritis patients age 21-78 years, take the pharmaceutical composition of the formula of the above example 1 orally 3 times a day with 3-4 tablets each time, each bag with 0.33mg, and take the composition for one to three treatment courses according to the severity of the patients. And (3) clinical test results: the traditional Chinese medicine composition can cure 10 patients, improves 9 patients, basically has no change for 1 patient, has the total effective rate of 95 percent, has stable curative effect, and does not relapse, depend on drug properties and toxic or side effect of the patients.

20 patients with upper respiratory tract infection, age 14-68 years, take the pharmaceutical composition of the formula of the above example 1 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take one to three courses of treatment according to the severity of the disease. And (3) clinical test results: the traditional Chinese medicine composition can cure 10 patients, improves 10 patients, basically has no change of 0 patient, has 100 percent of total effective rate, stable curative effect, no relapse of the cured patients, and no dependence on drug property and toxic or side effect.

25 bacillary dysentery patients, 25-71 years old, take the pharmaceutical composition of the formula of example 1 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take one to three courses of treatment according to the severity of the disease. And (3) clinical test results: 15 patients are cured, 10 patients are improved, 0 patient is basically not changed, the total effective rate is 100 percent, the curative effect is stable, and the cured patients do not relapse and do not depend on drug property and toxic or side effect.

Case 2:

24 patients with chronic gastritis are 18-70 years old, and the pharmaceutical composition prepared in the formula of the embodiment 2 is orally taken 3 times a day, 3-4 tablets are taken each time, 0.33mg is packed each time, and the pharmaceutical composition is taken for one to three treatment courses according to the severity of the illness of the patients. And (3) clinical test results: the traditional Chinese medicine composition can cure 12 patients, improves 10 patients, basically has no change for 2 patients, has the total effective rate of 91.67 percent, has stable curative effect, and has no relapse, drug property and toxic or side effect for the cured patients.

30 patients with acute gastritis, age 17-65 years old, take the pharmaceutical composition of the formula of the above example 2 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take one to three courses of treatment according to the severity of the patient's condition. And (3) clinical test results: 16 patients are cured, 10 patients are improved, 4 patients are basically not changed, the total effective rate is 87%, the curative effect is stable, and the cured patients do not relapse and do not depend on drug property and toxic or side effect.

Case 3:

24 patients with pharyngitis, the age of which is 20-71 years old, take the pharmaceutical composition prepared in the above example 3 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take the pharmaceutical composition for one to three treatment courses according to the severity of the patients. And (3) clinical test results: 11 patients are cured and improved, the total effective rate is 91.6 percent, the curative effect is stable, and the cured patients do not relapse, and are free from drug property and toxic and side effects.

19 patients with acute gastritis are aged 15-63 years, the pharmaceutical composition prepared in the formula in the embodiment 3 is orally taken 3 times a day, 3-4 tablets are taken each time, and each bag contains 0.33mg, and the pharmaceutical composition is taken for one to three treatment courses according to the severity of the patient. And (3) clinical test results: 11 patients are cured, 7 patients are improved, 1 patient basically has no change, the total effective rate is 94.7 percent, the curative effect is stable, and the cured patients have no relapse, drug property and toxic or side effect.

21 cases of acute gastroenteritis, age 18-70 years, adopt the above-mentioned formulation pharmaceutical composition of example 3, take orally, 3 times a day, 3-4 tablets each time, each bag is 0.33mg, according to the severity of the patient's disease, take one to three courses of treatment. And (3) clinical test results: the traditional Chinese medicine composition can cure 12 patients and improve 8 patients, basically has no change for 1 patient, has the total effective rate of 95.2 percent, has stable curative effect, and does not relapse, depend on drug properties and toxic or side effect for the cured patients.

Case 4:

25 patients with chronic pharyngitis aged 19-70 years old take the pharmaceutical composition of the formula of the above example 4 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take the composition for one to three treatment courses according to the severity of the patients. And (3) clinical test results: 11 patients are cured and improved, and the total effective rate is 88 percent, the curative effect is stable, and the cured patients do not relapse, and do not depend on drug property and toxic or side effect, and 3 patients are basically not changed.

26 patients with acute gastritis, age 13-65 years, take the pharmaceutical composition of the formula of the above example 4 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take one to three courses of treatment according to the severity of the patient's condition. And (3) clinical test results: 12 patients are cured and improved, the total effective rate is 92.3 percent, the curative effect is stable, and the cured patients do not relapse, and are free from drug property and toxic and side effects.

20 patients with acute gastroenteritis, age 21-73 years old, take the pharmaceutical composition of the above example 4 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take one to three courses of treatment according to the severity of the patients. And (3) clinical test results: the traditional Chinese medicine composition can cure 10 patients, improves 9 patients, basically has no change for 1 patient, has the total effective rate of 95 percent, has stable curative effect, and does not relapse, depend on drug properties and toxic or side effect of the patients.

18 cases of bacillary dysentery patients are aged 45-70 years, the pharmaceutical composition prepared in the above example 4 is orally taken 3 times a day, 3-4 tablets each time, 0.33mg each bag, and the patient takes a course of treatment according to the severity of the disease. And (3) clinical test results: 9 patients are cured and improved by 8 patients, 1 patient basically has no change, the total effective rate is 94.4 percent, the curative effect is stable, and the cured patients have no relapse, drug property and toxic or side effect.

Case 5:

27 patients with chronic pharyngitis aged 15-55 years old take the pharmaceutical composition of the formula of the example 5 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take the pharmaceutical composition for one to three treatment courses according to the severity of the disease. And (3) clinical test results: the traditional Chinese medicine composition can cure 12 patients, improves 11 patients, basically has no change of 4 patients, has 85 percent of total effective rate, stable curative effect, no relapse of cured patients, no dependence on drug property and toxic or side effect.

19 patients with acute gastritis, 20-63 years old, take the pharmaceutical composition of the formula of the above example 5 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take the composition for one to three treatment courses according to the severity of the patient's condition. And (3) clinical test results: the traditional Chinese medicine composition can cure 10 patients, improves 8 patients, basically has no change for 1 patient, has the total effective rate of 94 percent, has stable curative effect, and does not relapse, depend on drug properties and toxic or side effect.

30 patients with acute gastroenteritis, age 21-73 years old, take the pharmaceutical composition of the formula of example 5 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and take one to three courses of treatment according to the severity of the patients. And (3) clinical test results: 13 patients are cured, 7 patients are improved, 3 patients are basically not changed, the total effective rate is 90%, the curative effect is stable, and the cured patients do not relapse and do not depend on drug properties and toxic or side effects.

26 bacillary dysentery patients with the age of 35-67 years take the pharmaceutical composition of the formula of example 5 orally 3 times a day, 3-4 tablets each time, 0.33mg each bag, and a course of treatment is taken according to the severity of the disease. And (3) clinical test results: 11 patients are cured, 8 patients are improved, 2 patients are basically not changed, the total effective rate is 92.3 percent, the curative effect is stable, and the cured patients do not relapse, and do not depend on drug property and toxic or side effect.

The above examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention, which is intended to be covered by the claims and any design similar or equivalent to the scope of the invention.

Claims

1. The anti-inflammatory and antibacterial pharmaceutical composition is characterized in that the pharmaceutical composition comprises the following extracts of raw materials in parts by weight: 130-190 parts of pomegranate bark, 120-160 parts of scutellaria baicalensis, 100-140 parts of garden burnet and 100-140 parts of golden cypress.

2. The anti-inflammatory and antibacterial pharmaceutical composition as claimed in claim 1, wherein the active ingredients comprise the following extracts of raw materials in parts by weight: 150-170 parts of pomegranate bark, 130-150 parts of scutellaria baicalensis, 110-130 parts of garden burnet and 110-130 parts of golden cypress.

3. The anti-inflammatory and antibacterial pharmaceutical composition as claimed in claim 1, wherein the active ingredients comprise the following extracts of raw materials in parts by weight: 160 parts of pomegranate bark, 140 parts of scutellaria baicalensis, 120 parts of garden burnet and 120 parts of golden cypress.

4. A process for the preparation of an anti-inflammatory and antibacterial pharmaceutical composition according to any one of claims 1 to 3, comprising the steps of:

1) weighing the raw material components according to the designed weight parts;

5. The method for preparing an anti-inflammatory and antibacterial pharmaceutical composition according to claim 4, wherein the heating reflux extraction in step 2) is as follows: extracting with 6 times of 60% ethanol for 15min, extracting with 4 times of 60% ethanol for 10min, and mixing the filtrates.

6. The method for preparing an anti-inflammatory and antibacterial pharmaceutical composition as claimed in claim 4, wherein the two extraction methods of Scutellariae radix, radix Sangusorbae and cortex Phellodendri in step 3) are as follows: adding 60% ethanol 6 times the total weight of Scutellariae radix, radix Sangusorbae and cortex Phellodendri, extracting for 1 hr, filtering, adding 60% ethanol 4 times the total weight of Scutellariae radix, radix Sangusorbae and cortex Phellodendri into the residue, extracting for 0.5 hr, filtering, and mixing the filtrates.

7. The method for preparing an anti-inflammatory and antibacterial pharmaceutical composition as claimed in claim 4, wherein the mixed extract of step 4) is concentrated to a relative density of 1.25 and then spray-dried.

8. The method for preparing an anti-inflammatory and antibacterial pharmaceutical composition according to claim 4, further comprising step 5), adding crospovidone and sodium carboxymethyl starch as excipients into the concentrated and dried pharmaceutical composition of step 4) as disintegrants, microcrystalline cellulose as filler, mixing well, making soft mass with 60% by volume ethanol solution, granulating, drying at 50 ℃ to obtain granules, controlling the relative humidity of air below 66%, and compressing into dispersible tablets.

9. The preparation method of the anti-inflammatory and antibacterial pharmaceutical composition according to claim 8, wherein the mass ratio of the pharmaceutical composition to the auxiliary materials is 100-20: 254-350, and the mass ratio of crospovidone, sodium carboxymethyl starch and microcrystalline cellulose in the auxiliary materials is 1:1: 1.

10. Use of the anti-inflammatory and antibacterial pharmaceutical composition of any one of claims 1 to 3 in the preparation of a medicament for the treatment of inflammation.