CN113925862A

CN113925862A - Pharmaceutical composition for preventing and/or treating inflammatory storm and preparation and application thereof

Info

Publication number: CN113925862A
Application number: CN202111546423.XA
Authority: CN
Inventors: 刘志华; 刘玉玲; 刘璐; 冯遇; 姜丹; 刘瑾如; 王丽; 叶军; 邹媛媛
Original assignee: Guangxi Wuhe Boao Pharmaceutical Co ltd; Beijing Wuhebao Pharmaceutical Co ltd
Current assignee: Guangxi Wuhe Boao Pharmaceutical Co ltd; Beijing Wuhebao Pharmaceutical Co ltd
Priority date: 2021-12-17
Filing date: 2021-12-17
Publication date: 2022-01-14

Abstract

The invention discloses a pharmaceutical composition for preventing and/or treating inflammatory storm, a preparation and application thereof. The pharmaceutical composition consists of baicalein, wogonin and oroxylin A, wherein the mass ratio of the baicalein to the wogonin to the oroxylin A is 0.25-1.5: 0.5-7: 1. the self-microemulsion provided by the invention comprises the following components: a drug phospholipid complex, an oil phase, an emulsifier and a co-emulsifier; wherein, the medicine phospholipid compound is formed by compounding the medicine composition and a phospholipid material. The experimental result of the invention shows that in a mouse model of systemic inflammation storm induced by LPS, the composition of baicalein, wogonin and oroxylin A and the self-microemulsion preparation thereof have the function of inhibiting the mouse cytokine storm to different degrees. The invention provides a safe, effective and economic solution for the clinical treatment of the inflammatory storm.

Description

Pharmaceutical composition for preventing and/or treating inflammatory storm and preparation and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a pharmaceutical composition for preventing and/or treating inflammatory storm, and a preparation and application thereof.

Background

Inflammatory storm is an excessive immune and inflammatory response of the body to external stimuli, characterized by T cell and macrophage hyperproliferation and rapid release and overexpression of inflammatory cytokines. Inflammatory storms may be caused by a number of infectious or non-infectious diseases, such as influenza, new crown pneumonia (COVID-19), middle east respiratory syndrome (MERS-CoV), atypical pneumonia (SARS), familial hemophagocytic syndrome, sepsis, systemic onset idiopathic arthritis (sJIA), Chimeric Antigen Receptor (CAR) -T cell therapy, lupus erythematosus, and the like.

The inflammatory storm phenomenon caused by respiratory virus infection such as influenza virus and coronavirus is the most common phenomenon in clinic. Chaolin Huang et al report that the plasma concentration of proinflammatory cytokines such as IL-1 beta, IL-1RA, IL-7, IL-8, IL-9, FGF, G-CSF, GM-CSF, IFN-gamma, IP-10, MCP-1, MIP-1 alpha, MIP-1 beta, PDGF, TNF-alpha and VEGF were significantly increased in 2019-nCoV infected patients compared to healthy people, while the plasma concentrations of IL-2, IL-7, GSCF, IP-10, MCP-1, MIP-1 alpha and TNF-alpha were also significantly higher in severe patients entering the ICU than in non-severe patients [ reference 1: clinical features of tissues fed with 2019 novel coronavirus in Wuhan, China, The Lancet, 2020, 395(10223): 497-506 ].

The occurrence of inflammatory storms is closely related to the severity of the disease and the occurrence of multiple organ dysfunction syndrome. Aiming at 2019-nCoV infection, professor Zhouqi of hospital department of China suggests that the inflammatory storm is an important node for the conversion of new coronary patients from mild to severe and severe cases, and is also a cause of death of severe and severe patients. Reducing the incidence of inflammatory storms in infectious or non-infectious diseases helps to reduce organ damage and slow the progression of the disease, especially in critically ill patients. In the treatment strategy of acute influenza, studies have demonstrated that reducing the incidence of inflammatory storms using immunomodulators can effectively reduce the mortality and organ damage of acute influenza [ document 2: the cytokine storm of segment infiltration and degradation of Immunology therapy, Cellular & Molecular Immunology, 2016, 13: 3-10.

For the treatment strategy of the inflammatory storm, no specific medicine is available at present, and anti-infective drugs, glucocorticoids, peroxisome proliferator-activated receptor agonists, Cyclooxygenase (COX) inhibitors, antioxidants, TNF inhibitors, IL-1 antagonists, IL-6 antagonists and other methods are mostly adopted, so that the treatment effect is limited and the side effect is great. For example, glucocorticoids, while non-specifically inhibiting inflammatory storms, are prone to secondary infections, diabetes, osteoporosis, hypertension, osteonecrosis, and other side effects when administered for long periods. Therefore, the development of safe and effective new drugs for preventing and treating inflammatory storm has important significance for saving the life of patients.

The traditional Chinese medicine has multiple target points and wide effect, is mainly systemically and integrally regulated, and has higher value in the prevention, treatment and conditioning of complex diseases. The scutellaria is a common traditional Chinese medicine, has more than two thousand years of medicinal history in China, and is clinically used for diseases such as fever, upper respiratory tract infection, pneumonia, acute gastroenteritis, dysentery, hepatitis and the like. The flavonoids are the main drug effect substance basis of radix Scutellariae, and comprise baicalin, wogonoside, baicalein, wogonin, oroxylin A, etc. However, the application of the baicalein active ingredient in the preparation of the medicament for preventing and treating the inflammation storm is not reported at present.

Disclosure of Invention

It is a first object of the present invention to provide a pharmaceutical composition for preventing and/or treating inflammatory storm.

The pharmaceutical composition for preventing and/or treating the inflammation storm provided by the invention comprises baicalein, wogonin and oroxylin A, wherein the mass ratio of the baicalein to the wogonin to the oroxylin A is 0.25-1.5: 0.5-7: 1.

further, the mass ratio of the baicalein, the wogonin and the oroxylin A is (0.5-1.5): 5-7): 1; or the mass ratio of the baicalein, the wogonin and the oroxylin A is (0.29-0.43): 0.57-0.86): 1.

According to an embodiment of the present invention, the composition may have a ratio of baicalein, wogonin, oroxylin a of 0.5:7:1 by mass.

According to an embodiment of the present invention, in the composition, the mass ratio of the baicalein, the wogonin and the oroxylin A can be 1.5:5: 1.

According to an embodiment of the present invention, in the composition, the mass ratio of the baicalein, the wogonin and the oroxylin A may be 0.43:0.57: 1.

according to an embodiment of the present invention, in the composition, the mass ratio of the baicalein, the wogonin and the oroxylin A may be 0.29:0.86: 1.

in the invention, the baicalein can be baicalein raw material medicine, salt or baicalein in different crystal forms.

In the invention, the wogonin can be wogonin bulk drug, salt or wogonin in different crystal forms.

In the invention, the oroxylin A can be oroxylin A raw material medicine, salt or oroxylin A in different crystal forms.

The second purpose of the invention is to provide a medicament for preventing and/or treating inflammatory storm.

The active ingredients of the medicine for preventing and/or treating the inflammatory storm provided by the invention comprise the pharmaceutical composition.

Further, the medicine also comprises a pharmaceutically acceptable carrier or auxiliary material.

Furthermore, the medicine can also comprise pharmaceutically acceptable medicines for preventing and treating inflammatory storm or other related active substances.

Further, the dosage form of the medicament comprises an oral preparation, an injection preparation, a transdermal administration preparation, a mucosa administration preparation, a lung inhalation administration preparation or an intestinal administration preparation;

the specific dosage forms are as follows: drop, oral liquid, tablet, capsule, granule, pellicle, gel, powder, emulsion, self-emulsifying preparation, dripping pill, suppository, aerosol, spray, powder spray, patch, solution, unguent or cream.

The third purpose of the invention is to provide a self-microemulsion containing the drug composition and a preparation method thereof.

The self-microemulsion provided by the invention comprises the following components: a drug phospholipid complex, an oil phase, an emulsifier and a co-emulsifier;

wherein, the medicine phospholipid compound is formed by compounding the medicine composition and a phospholipid material; the mass content of the pharmaceutical composition is 5-75%, preferably 10-30%, more preferably 15-25%; the mass content of the phospholipid is 25-95%, preferably 70-90%, more preferably 75-85%, and the mass content of the pharmaceutical composition refers to the percentage of the pharmaceutical composition in the pharmaceutical phospholipid complex.

Wherein the phospholipid material is natural phospholipid or synthetic phospholipid or their mixture, specifically phospholipid S75.

According to one embodiment of the invention, the pharmaceutical phospholipid complex is prepared by mixing the pharmaceutical composition and the phospholipid material according to a mass ratio of 1: 3.5 in proportion.

The preparation method of the drug phospholipid complex provided by the invention comprises the following steps: weighing the components in the medicinal composition according to the mass ratio, weighing the medicinal composition and the phospholipid material according to the medicinal-lipid ratio, placing the medicinal composition and the phospholipid material in a rotary evaporation bottle, adding a proper amount of reaction solvent, mixing and shaking uniformly, standing for 15-30 min after the medicinal composition and the phospholipid are completely dissolved, placing the medicinal composition on a rotary evaporation instrument, volatilizing the solvent, drying under reduced pressure and vacuum, and scraping solids to obtain the medicinal phospholipid complex.

In the above method, the reaction solvent may be selected from anhydrous ethanol.

In the method, the temperature of the solvent volatilizing can be 40 ℃, and the time of the reduced pressure vacuum drying is more than 12 h.

In the present invention, the oil phase is selected from at least one of: ethyl oleate, isopropyl myristate, caprylin, castor oil, Maisine, medium chain triglycerides, preferably from at least one of the following: ethyl oleate, isopropyl myristate, caprylin; more preferably ethyl oleate;

the emulsifier is selected from at least one of the following: kolliphor RH40, Kolliphor EL, Tween 85, Tween 80, Labrasol, PLURL QLEIQUE CC497, X-100, preferably selected from: tween 80, Labrasol or tween 80 mixed with Labrasol emulsifier, more preferably tween.

The coemulsifier is selected from at least one of the following: PEG400, absolute ethanol, propylene glycol, n-butanol, Capryol 90, glycerol, Transcutol HP, preferably selected from: transcutol HP or Capryol 90, more preferably Transcutol HP.

In the self-microemulsion, the mass ratio of the oil phase to the emulsifier to the co-emulsifier is 1: (0.5-8): (0.5-8), preferably 2:5: 3.

In the self-microemulsion, the content of the drug phospholipid compound is 10-50%, preferably 10% of the total mass of the oil phase, the emulsifier and the co-emulsifier.

According to one embodiment of the invention, the self-microemulsion comprises the following components: a drug phospholipid complex, ethyl oleate, tween 80 and Transcutol HP; wherein the mass ratio of the ethyl oleate to the Tween 80 to the Transcutol HP is 2:5:3, and the dosage of the medicine phospholipid compound is 10% of the sum of the mass of the ethyl oleate, the mass of the Tween 80 and the mass of the Transcutol HP.

The preparation method of the drug phospholipid complex self-microemulsion provided by the invention comprises the following steps: weighing the oil phase, the emulsifier and the co-emulsifier according to the proportion, and uniformly shaking to prepare a blank self-microemulsion solution; weighing a proper amount of the medicinal phospholipid compound, placing the medicinal phospholipid compound into the blank self-microemulsion, placing the medicinal phospholipid compound into an air bath oscillator, and oscillating and dissolving completely to obtain the medicinal phospholipid compound self-microemulsion.

Wherein the set temperature of the air bath oscillator is 25 ℃ and the rotating speed is 210 rpm.

The fourth purpose of the invention is to provide the medicinal composition, the medicament and the application of the self-microemulsion.

The application provided by the invention is the application of the pharmaceutical composition, the medicine or the self-microemulsion in preparing the medicine for preventing and/or treating the inflammatory storm.

The inflammatory storm is a phenomenon that a plurality of cytokines in a body are rapidly and massively produced due to infection or non-infectious diseases and tissue inflammatory injury caused by the cytokines.

The infectious disease may be an infection by a virus, chlamydia, mycoplasma, bacteria or parasite.

The non-infectious disease can be autoimmune hepatitis, arthritis, pancreatitis, gastroenteritis, familial hemophagocytic syndrome, systemic juvenile idiopathic arthritis, Chimeric Antigen Receptor (CAR) -T cell therapy, lupus erythematosus, and the like.

The various cytokines in the body include one or more of interleukins, interferons, growth factors, tumor necrosis factors, chemokines and colony stimulating factors.

The various cytokines in the body include one or more of MCP-1, IFN-beta, IL-1 beta, IL-23, IL-10, IL-12p70, MIG, IP-10, MIP-1 alpha, TNF-alpha, IL-27, VEGF, IL-1 alpha, GM-CSF, MIP-1 beta, IL-4, IFN-gamma, IL-6, IL-17A.

The tissue inflammatory injury comprises lung injury, liver injury, kidney injury, heart injury, stomach injury, intestinal injury, etc.

The inflammatory storm in the invention can be specifically a sepsis systemic inflammatory storm.

The experimental result of the invention shows that in a mouse model of systemic inflammation storm induced by LPS, the composition of baicalein, wogonin and oroxylin A and the self-microemulsion preparation thereof have the function of inhibiting the mouse cytokine storm to different degrees. The invention provides a safe, effective and economic solution for the clinical treatment of the inflammatory storm.

Drawings

Fig. 1 is the mean plasma concentration of each factor in each dosing group after 1.5h after LPS induction in example 3, 1-SMEDSS to 4-SMEDSS represent ratios 1 to 4, respectively, from the microemulsion, groups 1 to 4 represent ratios 1 to 4, respectively, of the bulk drug composition, and groups represent p <0.001, p <0.0001, compared to the LPS group (n = 5);

fig. 2 shows the mean plasma concentrations of the factors in each group administered 4h after LPS induction in example 3, 1-SMEDSS to 4-SMEDSS represent the ratios 1 to 4 self-microemulsions, respectively, and groups 1 to 4 represent the ratios 1 to 4 bulk drug compositions, respectively, and —. represents p <0.001,. p <0.0001, compared to the LPS group (n =5,. p represents p <0.05,. p represents p < 0.01);

fig. 3 is the mean plasma concentrations of IFN- β, IFN- γ in each group administered 4h after LPS induction in example 3, 1-SMEDSS to 4-SMEDSS represent the ratios 1 to 4 self-microemulsions, respectively, and groups 1 to 4 represent the ratios 1 to 4 drug substance compositions, respectively, (n =5, p <0.05, p <0.01, p <0.001, p < 0.0001);

fig. 4 shows the mean plasma concentrations of the factors in each group administered 6h after LPS induction in example 3, 1-SMEDSS to 4-SMEDSS represent the ratios 1 to 4 self-microemulsions, respectively, and groups 1 to 4 represent the ratios 1 to 4 drug substance compositions, respectively, and these values represent p <0.001, p <0.0001, compared to the LPS group (n = 5).

Detailed Description

The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.

Example 1 preparation of drug phospholipid Complex

Weighing 1g of the medicinal composition (wherein the mass ratio of baicalein to wogonin to oroxylin A is 0.5:7:1 (ratio 1), 1.5:5:1 (ratio 2), 0.29:0.86:1 (ratio 3) and 0.43:0.57:1 (ratio 4) of phospholipid S753.5 g according to the medicinal-lipid ratio (1: 3.5, w/w), placing the medicinal composition in a rotary steaming bottle, adding 20mL of tetrahydrofuran, mixing, shaking uniformly, standing for 30min after the raw material medicaments and the phospholipid are completely dissolved, placing the medicinal composition on a rotary steaming instrument, volatilizing the solvent at 40 ℃, drying under reduced pressure for more than 12h, and scraping solids to obtain medicinal phospholipid complexes with different medicinal proportions.

Example 2 preparation of self-microemulsions

Weighing ethyl oleate, Tween 80 and Transcutol HP according to the mass ratio of 2:5:3, uniformly shaking to prepare a blank self-microemulsion solution, adding the medicinal phospholipid compound prepared in the example 1 into the blank self-microemulsion solution, wherein the feeding amount is 10% (taking the blank emulsion as 100%) and placing the mixture into an air bath oscillator, setting the temperature at 25 ℃, the rotating speed at 210 rpm, and completely oscillating and dissolving to obtain the self-microemulsions with different medicament ratios.

Example 3 pharmacodynamic experiment of pharmaceutical composition and self-microemulsion

(1) Experimental Material

Baicalein crude drug (purity 98%, Nanjing Zelang Biotech Co., Ltd., batch number: ZLS 1903601);

wogonin drug substance (batch No. 111514-201706, China institute for food and drug testing), oroxylin A drug substance (batch No. PRF10030442, Chengdu-pu-Rui-Tech-Co., Ltd.), Lipopolysaccharides from Escherichia coli 055: B5 (SIGMA); sodium chloride injection (Sichuan Kelun pharmaceutical Co., Ltd.); mouse IL-6 ELISA Kit (Elapscience); mouse Inmigration Panel, Mouse Cytokine Release Syndrome Panel, Mouse Proinfringement Chemokine Panel (Biolegend); c57 BL/6 male mouse, 18-22g, (Beijing Huafukang Biotech GmbH); one hundred thousand electronic balance (METTLER TOLEDO) type XS 105 DU; model 1-14 centrifuges (SIGMA); VORTEX-2 GENIE model VORTEX apparatus (Scientific Industries); model C6 Flow cytometer (BD Accuri).

(2) Experiment grouping

After adaptive feeding of C57 BL/6 male mice, the mice were randomly divided into a normal group, a model group, a baicalein group, a composition ratio 1 solution group, a composition ratio 2 solution group, a composition ratio 3 solution group, a composition ratio 4 solution group, a composition ratio 1 self-microemulsion group, a composition ratio 2 self-microemulsion group, a composition ratio 3 self-microemulsion group and a composition ratio 4 self-microemulsion group, wherein 18 mice were each group, and the mice were fasted for 12h before the experiment without water prohibition. The normal group and the model group are orally administrated with normal saline with equal amount by gavage, the composition (suspended in 0.1% sodium carboxymethyl cellulose aqueous solution) mixed by the mass ratio of 0.5:7:1 of baicalein, wogonin and oroxylin is orally administrated with gavage by the composition ratio of 1 solution group, the composition (suspended in 0.1% sodium carboxymethyl cellulose aqueous solution) mixed by the mass ratio of 1.5:5:1 of baicalein, wogonin and oroxylin is orally administrated with gavage by the composition ratio of 2 solution group, the composition (suspended in 0.1% sodium carboxymethyl cellulose aqueous solution) mixed by the mass ratio of 1.5:5:1 of baicalein, wogonin and oroxylin is orally administrated with the composition ratio of 3 solution group, the composition (suspended in 0.1% sodium carboxymethyl cellulose aqueous solution) is 80mg/kg mixed by the mass ratio of 0.29:0.86:1 of baicalein and the composition ratio of 4 solution group is orally administrated with baicalein, the composition ratio of 4 solution group is orally administrated with gavage by the baicalein, the combination ratio of baicalein and the combination is orally administrated, 80mg/kg of a composition (suspended in 0.1% sodium carboxymethylcellulose aqueous solution) in which wogonin and oroxylin are mixed in a mass ratio of 0.43:0.57:1, 80mg/kg of baicalein, wogonin, and oroxylin which are prepared in example 2 were orally gavage in a composition ratio of 1 and are mixed in a mass ratio of 0.5:7:1, 80mg/kg of self-microemulsion in which baicalein, wogonin, and oroxylin are mixed in a mass ratio of 1.5:5:1, 3 of baicalein, wogonin, and oroxylin which are prepared in example 2 were orally gavage in a composition ratio of 2, 80mg/kg of self-microemulsion in which are mixed in a mass ratio of 0.29:0.86:1, 4 of composition, 4 of baicalein, and oroxylin which are orally administered in a microemulsion ratio of 0.43:0.57:1, The self-microemulsion is 80mg/kg of wogonin and oroxylin which are mixed according to the mass ratio of 0.43:0.57: 1. The administration dose of 80mg/kg for each of the above groups means the dose of the active ingredient.

After 40 min, the model group, the composition solution group in each proportion and the composition self-microemulsion group are all injected with LPS (5 mg/kg) in the abdominal cavity to induce the systemic inflammatory storm, and the normal group is injected with normal saline with the same volume in the abdominal cavity. After 1.5h, 4h and 6h of LPS injection, 6 mice were bled from the eyeball at each blood sampling point in each administration group. Collecting blood with blood collecting tube, centrifuging at 4000rpm/min for 10min at 2-8 deg.C within 30min, collecting supernatant, packaging plasma, and storing at-20 deg.C.

(3) Determination of inflammatory cytokine concentration in plasma

Cytokine concentrations in Mouse plasma were determined according to Mouse infection Panel, Mouse Cytokine Release Synthesis Panel, Mouse Proinfringement Chemokine Panel and the methods indicated in the instructions in the ELISA kits.

(4) Results of the experiment

Data analysis was performed using Graphpad prism9.0 software, and the data were expressed by Mean + -SEM, and the results are shown in FIGS. 1-3.

As shown in FIGS. 1 to 4, after lipopolysaccharide is injected into the abdominal cavity of a mouse, the concentration of the lipopolysaccharide in blood plasma is remarkably increased, which indicates that the model is successfully established.

As shown in figure 1, after 1.5h after LPS induction, the ratio 1 self-microemulsion (0.5: 7: 1) can obviously inhibit IP-10 and MIP-1 alpha in the plasma of a model mouse, the ratio 2 self-microemulsion (1.5: 5: 1) can obviously inhibit IP-10 and MIP-1 alpha in the plasma of the model mouse, the ratio 3 self-microemulsion (0.29: 0.86: 1) can obviously inhibit IP-10, MIP-1 alpha, MCP-1, IL-6 and IL-12p70 in the plasma of the model mouse, the ratio 4 self-microemulsion (0.43: 0.57: 1) can obviously inhibit IP-10, MIP-1 alpha, MCP-1, IL-6, GM-CSF and IL-12p70 in the plasma of the model mouse, the ratio 1 bulk drug (0.5: 7: 1) can obviously inhibit IFN-gamma in the plasma of the model mouse, and the ratio 2 bulk drug (1.5: 5: 1) can obviously inhibit IP-10 in the plasma of the model mouse, MIP-1 alpha, IFN-gamma, proportion 3 raw material medicine (0.29: 0.86: 1) can obviously inhibit IFN-gamma, IL-6, IL-1 alpha, IL-12p70 in model mouse plasma.

As above, the ratios and the ratios of the raw material drugs are the same, and are not repeated here, as shown in FIG. 2-3, 4 hours after LPS induction, the ratio 1 self-microemulsion can significantly inhibit GM-CSF, IL-1 beta and IL-17A, IFN-beta in the plasma of the model mouse, the ratio 2 self-microemulsion can significantly inhibit IP-10, IL-1 alpha, GM-CSF, IL-12p70, IL-1 beta, IL-6, IL-27, IL-17A, IFN-beta and IFN-gamma in the plasma of the model mouse, the ratio 3 self-microemulsion can significantly inhibit GM, IP-10, IL-1 alpha, GM-CSF, IL-12p70, IL-1 beta, IL-6, IL-27 and IL-17A, IFN-gamma in the plasma of the model mouse, and the ratio 4 self-microemulsion can significantly inhibit GM, IL-1 beta, IL-17, gamma in the plasma of the model mouse, IP-10, IL-17A, IFN-beta and IFN-gamma, the raw material medicine with the proportion of 1 can obviously inhibit GM-CSF, IL-17A, IFN-beta and IFN-gamma in the blood plasma of a model mouse, the raw material medicine with the proportion of 2 can obviously inhibit IP-10, IL-1 alpha and IL-17A, IFN-beta in the blood plasma of the model mouse, the raw material medicine with the proportion of 3 can obviously inhibit IFN-beta and IFN-gamma in the blood plasma of the model mouse, and the raw material medicine with the proportion of 4 can obviously inhibit IP-10 and IL-17A, IFN-beta in the blood plasma of the model mouse.

As shown in figure 4, after 6h of LPS induction, the self-microemulsion with the proportion 1 can obviously inhibit IL-27 and IL-1 beta in the plasma of a model mouse, the self-microemulsion with the proportion 2 can obviously inhibit IFN-gamma, IL-17A, GM-CSF, TNF-alpha, IL-1 beta and MCP-1 in the plasma of the model mouse, the self-microemulsion with the proportion 3 can obviously inhibit TNF-alpha and IL-1 beta in the plasma of the model mouse, the self-microemulsion with the proportion 4 can obviously inhibit TNF-alpha in the plasma of the model mouse, the bulk drug with the proportion 2 can obviously inhibit TNF-alpha and IL-1 beta in the plasma of the model mouse, and the bulk drug with the proportion 4 can obviously inhibit IFN-gamma, IL-27 and IL-17A, IL-1 beta in the plasma of the model mouse.

Meanwhile, the inhibition rate of 1.5h-6h self-microemulsion and raw material medicaments to each inflammatory factor is detected, and the result is as follows,

inhibition ratio (%) = | administration group factor concentration-LPS group factor concentration |/LPS group factor concentration ×. 100%

The ratio 1 self-microemulsion (0.5: 7: 1) can obviously inhibit the increase of the concentration of 7 inflammatory cytokines such as IP-10, MIP-1 alpha, IL-1 beta, IL-17A, GM-CSF, IL-27, IFN-beta and the like in the plasma of a model mouse, and the inhibition rates are 58.58%, 76.03%, 36.79%, 54.36%, 40.47%, 64.08% and 97.15% respectively.

The ratio 2 self-microemulsion (1.5: 5: 1) can obviously inhibit the increase of the concentrations of 13 inflammatory cytokines such as IP-10, MIP-1 alpha, IL-1 alpha, IFN-gamma, IL-12p70, IL-1 beta, IL-6, IL-17A, GM-CSF, IL-27, TNF-alpha, MCP-1, IFN-beta and the like in the plasma of a model mouse, and the inhibition rates are 57.85%, 67.92, 84.25, 99.69, 84.72, 81.97, 73.67, 88.46, 80.68, 96.40, 99.86, 80.14 and 96.36 respectively.

The ratio 3 self-microemulsion (0.29: 0.86: 1) can obviously inhibit the increase of the concentrations of 13 inflammatory cytokines such as IP-10, MIP-1 alpha, MCP-1, IL-12p70, IL-6, MIG, IL-1 alpha, IFN-gamma, IL-1 beta, IL-27, IL-17A, GM-CSF, TNF-alpha and the like in the plasma of a model mouse, and the inhibition rates are respectively 97.37%, 73.66%, 88.09%, 77.41%, 85.19%, 92.86%, 62.16%, 81.34%, 35.17%, 77.26%, 65.65% and 60.63%.

The ratio 4 self-microemulsion (0.43: 0.57: 1) can obviously inhibit the increase of the concentrations of 11 inflammatory cytokines such as IP-10, MIP-1 alpha, MCP-1, IL-12p70, IL-6, GM-CSF, MIG, IFN-gamma, IL-17A, TNF-alpha, IFN-beta and the like in the plasma of a model mouse, and the inhibition rates are 76.77%, 91.39%, 71.67%, 93.19%, 97.67%, 78.55%, 59.35%, 32.49%, 54.32%, 71.03% and 94.17%, respectively.

The raw material medicine with the proportion of 1 (0.5: 7: 1) can obviously inhibit the increase of the concentrations of 4 inflammatory cytokines such as IFN-gamma, IL-17A, GM-CSF, IFN-beta and the like in the plasma of a model mouse, and the inhibition rates are 57.65%, 43.83%, 31.06% and 94.77% respectively.

The raw material medicine with the ratio 2 (1.5: 5: 1) can obviously inhibit the increase of the concentrations of 8 inflammatory cytokines such as IP-10, MIP-1 alpha, IFN-gamma, IL-1 alpha, IL-17A, TNF-alpha, IL-1 beta, IFN-beta and the like in the plasma of a model mouse, and the inhibition rates are 65.78%, 68.94%, 58.89%, 58.68%, 38.52%, 89.49%, 35.24% and 89.99% respectively.

The raw material medicine with the proportion of 3 (0.29: 0.86: 1) can obviously inhibit the increase of the concentrations of 4 inflammatory cytokines such as IFN-gamma, IL-1 alpha, IL-6, IFN-beta and the like in the plasma of a model mouse, and the inhibition rates are 66.84%, 92.03%, 86.31% and 90.08% respectively.

The bulk drug with the ratio of 4 (0.43: 0.57: 1) can obviously inhibit the increase of the concentration of 6 inflammatory cytokines such as IP-10, IL-17A, IFN-beta, IFN-gamma, IL-1 beta, IL-27 and the like in the plasma of a model mouse, and the inhibition rates are 47.32%, 71.35%, 97.24%, 86.44%, 61.80% and 88.47% respectively.

By combining the above analysis results, it is possible to obtain: the compositions with different proportions and the self-microemulsion preparation thereof have the function of inhibiting the cytokine storm with different degrees. The self-microemulsion preparation has better inhibition effect than the solution group with the same proportion.

(5) The state of each group of mice is observed after 1.5-4 h of LPS (lipopolysaccharide) intraperitoneal injection as follows:

1) normal group: the mouse is excited, the body temperature is normal and the activity is full;

2) model group: the mice have the problems of four limbs contracture, body temperature reduction, dull, poor activity and mental hypokinesia;

3) composition ratio 1 solution group: the body temperature of the mouse is cool, the limbs are curled, and the activity is poor;

4) composition ratio 2 solution group: the body temperature of the mouse is slightly cool, the limbs of the mouse are slightly curled, the activity is good, and the spirit is slightly poor;

5) composition ratio 3 solution group: the body temperature of the mouse is cool, the limbs are curled, and the activity is poor;

6) composition ratio 4 solution group: the body temperature of the mouse is slightly cool, the limbs of the mouse are curled, and the mobility of the mouse is normal;

7) composition ratio 1 self-microemulsifying group: the body temperature of the mouse is slightly cool, the limbs are slightly curled, the activity is general, and the mental state is satisfactory;

8) composition ratio 2 self-microemulsifying group: the body temperature of the mouse is normal, the activity is good, and the mental state is good;

9) composition ratio 3 self-microemulsifying group: the body temperature of the mouse is normal, the activity is good, and the mental state is good;

10) composition ratio 4 self-microemulsifying group: the mouse has normal body temperature, good activity and good mental state.

Claims

1. A pharmaceutical composition for preventing and/or treating inflammation storm comprises baicalein, wogonin and oroxylin A, wherein the mass ratio of the baicalein to the wogonin to the oroxylin A is 0.25-1.5: 0.5-7: 1.

2. the pharmaceutical composition of claim 1, wherein:

the baicalein is selected from baicalein raw material medicine, salt or baicalein in different crystal forms;

the wogonin is selected from wogonin bulk drug, wogonin in salt or different crystal forms;

the oroxylin A is selected from oroxylin A raw material medicine, salt or oroxylin A in different crystal forms.

3. A medicament for preventing and/or treating inflammatory storm, which comprises the pharmaceutical composition of claim 1 or 2 as an active ingredient.

4. The medicament of claim 3, wherein: the medicine also comprises a pharmaceutically acceptable carrier or auxiliary material;

or/and the medicine can also comprise pharmaceutically acceptable medicines for preventing and treating the inflammatory storm or other related active substances.

5. The medicament according to claim 3 or 4, characterized in that: the dosage form of the medicine comprises an oral preparation, an injection preparation, a transdermal administration preparation, a mucosal administration preparation, a pulmonary inhalation administration preparation or an intestinal administration preparation;

the specific dosage form comprises: drop, oral liquid, tablet, capsule, granule, pellicle, gel, powder, emulsion, self-emulsifying preparation, dripping pill, suppository, aerosol, spray, powder spray, patch, solution, unguent or cream.

6. A self-microemulsion containing a pharmaceutical composition according to claim 1 or 2, comprising the following components: a drug phospholipid complex, an oil phase, an emulsifier and a co-emulsifier;

wherein, the drug phospholipid complex is formed by compounding the drug composition of claim 1 or 2 and phospholipid material; the mass content of the pharmaceutical composition is 5-75%, preferably 10-30%, more preferably 15-25%; the mass content of the phospholipid is 25-95%, preferably 70-90%, more preferably 75-85%; the phospholipid material is natural phospholipid or synthetic phospholipid or a mixture thereof.

7. The self-microemulsion according to claim 6, wherein:

the oil phase is selected from at least one of: ethyl oleate, isopropyl myristate, caprylin, castor oil, Maisine, medium chain triglycerides, preferably from at least one of the following: ethyl oleate, isopropyl myristate, caprylin; more preferably ethyl oleate;

the emulsifier is selected from at least one of the following: kolliphor RH40, Kolliphor EL, Tween 85, Tween 80, Labrasol, PLURL QLEIQUE CC497, X-100, preferably selected from: tween 80, Labrasol or a mixture of tween 80 and Labrasol as emulsifier, more preferably tween;

the coemulsifier is selected from at least one of the following: PEG400, absolute ethanol, propylene glycol, n-butanol, Capryol 90, glycerol, Transcutol HP, preferably selected from: transcutol HP or Capryol 90, more preferably Transcutol HP;

in the self-microemulsion, the mass ratio of the oil phase to the emulsifier to the co-emulsifier is 1: (0.5-8): (0.5-8), preferably 2:5: 3;

8. Use of a pharmaceutical composition according to claim 1 or 2 or a medicament according to any one of claims 3 to 5 or a self-microemulsion according to claim 6 or 7 for the preparation of a medicament for the prevention and/or treatment of inflammatory storms.

9. Use according to claim 8, characterized in that: the inflammatory storm is a phenomenon that a plurality of cytokines in a body are rapidly and massively produced due to infection or non-infectious diseases and tissue inflammatory injury caused by the cytokines.

10. Use according to claim 8 or 9, characterized in that: the infectious disease is infection caused by virus, chlamydia, mycoplasma, bacteria or parasite;

the non-infectious disease is autoimmune hepatitis, arthritis, pancreatitis, gastroenteritis, familial hemophagocytic syndrome, systemic juvenile idiopathic arthritis, Chimeric Antigen Receptor (CAR) -T cell therapy, and lupus erythematosus;

the various cytokines in the organism comprise one or more of interleukins, interferons, tumor necrosis factors, chemokines and colony stimulating factors; specifically selected from: one or more of MCP-1, IFN-beta, IL-1 beta, IL-23, IL-10, IL-12p70, MIG, IP-10, MIP-1 alpha, TNF-alpha, IL-27, VEGF, IL-1 alpha, GM-CSF;

the tissue inflammatory injury comprises at least one of: lung injury, liver injury, kidney injury, heart injury, stomach injury, and intestinal injury.