Cinnamaldehyde drug-loaded self-microemulsion as well as preparation method and application thereof

Abstract

The invention provides a cinnamyl aldehyde drug-loaded self-microemulsion as well as a preparation method and application thereof, and belongs to the technical field of biological pharmacy. The cinnamyl aldehyde drug-loaded self-microemulsion disclosed by the invention is prepared from the following raw materials in parts by weight: 0.5 to 5.5 parts of cinnamyl aldehyde, 0 to 5.5 parts of oil phase, 0.5 to 5.5 parts of emulsifier and 0.01 to 0.045 part of insoluble drug. By adopting the self-microemulsion drug delivery system, the drug effect of the cinnamyl aldehyde and the combination of the cinnamyl aldehyde and other traditional Chinese medicine components is improved, the drug loading capacity is high, dispersion is easy, the stability is high, the drug effect is remarkable after the cinnamyl aldehyde and other traditional Chinese medicine components are combined, and the curative effect is good. The traditional Chinese medicine composition has an improvement effect on hepatic fibrosis, has a remarkable treatment effect on non-alcoholic steatohepatitis, and has certain application and development values.

Description

Technical Field

[0001] The invention belongs to the technical field of biopharmaceuticals, and specifically relates to a cinnamaldehyde drug-loaded self-microemulsion and a preparation method and application thereof. Background Art

[0002] Patients with non-alcoholic fatty liver disease (NAFLD) show elevated triglycerides, elevated LDL, and decreased HDL in biochemical tests. These symptoms are usually associated with features of metabolic syndrome, such as obesity, dyslipidemia, type 2 diabetes, and hypertension. Hepatocyte fatty degeneration and lipid accumulation are the earliest stages of NAFLD, which are usually asymptomatic. Appropriate changes in diet or lifestyle can reverse the affected liver to a healthy state. Subsequently, fat toxicity, oxidative stress, endoplasmic reticulum stress, etc. cause liver cell damage, triggering a large amount of inflammation, and fatty liver turns into non-alcoholic steatohepatitis (NASH). If left untreated, continued liver damage and inflammatory processes will further activate hepatic stellate cells, secrete collagen, and form liver fibrosis. Simple fatty degeneration and NASH are both reversible stages of NAFLD, while the late stage of transformation into cirrhosis or even liver cancer can only be treated by liver transplantation and is irreversible. However, there are currently few therapeutic drugs for each stage of NAFLD. Therefore, the development of new anti-NAFLD drugs is urgent, especially for early and middle fatty liver, NASH, and liver fibrosis stages, which is of great significance.

[0003] Cinnamaldehyde (CIN) is an aldehyde organic compound extracted from cinnamon, a traditional medicinal and edible plant. It is also present in essential oils of plants such as Patchouli, Hyacinthus and Rose. It is a light yellow oily liquid at room temperature and has antibacterial, lipid-lowering and antioxidant effects. Costunolide and dehydrocostusolide are substances extracted from the root of Costus vulgaris of the Asteraceae family, which have smooth muscle relaxation and spasmolysis effects. Dihydrotanshinone I is an active substance from the traditional Chinese medicine Salvia miltiorrhiza, which has antibacterial, platelet aggregation inhibition, antioxidant, and coronary vasodilation activities. However, the clinical application of cinnamaldehyde, costunolide, dehydrocostusolide and dihydrotanshinone I is severely limited due to their low solubility, poor lipid solubility, poor gastrointestinal absorption and rapid metabolism. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a cinnamaldehyde drug-loaded self-microemulsion and a preparation method and application thereof. The cinnamaldehyde drug-loaded self-microemulsion improves the efficacy of cinnamaldehyde and the combination of cinnamaldehyde and other traditional Chinese medicine ingredients, and has a high drug loading amount, is easy to disperse and has good stability, has an improvement effect on liver fibrosis and has a significant therapeutic effect on non-alcoholic fatty hepatitis.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: The invention provides a cinnamaldehyde drug-carrying self-microemulsion, comprising the following raw materials in parts by weight: 0.5-5.5 parts of cinnamaldehyde, 0-5.5 parts of oil phase, 0.5-5.5 parts of emulsifier and 0.01-0.045 parts of poorly soluble drugs; the mass ratio of the cinnamaldehyde to the emulsifier is 1:0.9-3.5.

[0006] Preferably, the method comprises the following raw materials in parts by weight: 0.8-4.5 parts of cinnamaldehyde, 0-4.5 parts of oil phase, 0.8-4.5 parts of emulsifier and 0.01-0.035 parts of drug; the mass ratio of cinnamaldehyde to emulsifier is 1:0.9-1.1.

[0007] Preferably, the mass ratio of the oil phase to the emulsifier is 1:0.9-3.5.

[0008] Preferably, the oil phase comprises carbon chain triglycerides.

[0009] Preferably, the emulsifier includes one or more of castor oil polyoxyethylene ether and polyoxyethylene hydrogenated castor oil.

[0010] Preferably, the poorly soluble drug includes one or more of dihydrotanshinone I, dehydrocostus lactone, and costusinolone.

[0011] The invention provides a method for preparing the cinnamaldehyde drug-carrying self-microemulsion, comprising the following steps: uniformly mixing the cinnamaldehyde, oil phase, emulsifier and drug in weight parts to obtain the cinnamaldehyde drug-carrying self-microemulsion.

[0012] Preferably, the mixing rotation speed is 550-650 rpm.

[0013] The present invention provides the cinnamaldehyde drug-loaded self-microemulsion or the use of the cinnamaldehyde drug-loaded self-microemulsion obtained by the preparation method in preparing a product for improving liver fibrosis.

[0014] The present invention provides the use of the cinnamaldehyde drug-loaded self-microemulsion or the cinnamaldehyde drug-loaded self-microemulsion obtained by the preparation method in preparing a product for treating fatty hepatitis.

[0015] Compared with the prior art, the present invention has the following beneficial effects: The present invention adopts a self-microemulsification drug delivery system to improve the efficacy of cinnamaldehyde and cinnamaldehyde and other Chinese medicines after being used in combination. In addition, the cinnamaldehyde in the cinnamaldehyde-loaded self-microemulsion is not only used as a drug component, but also can be used as the oil phase of the self-microemulsion. Compared with a single nanoemulsion system, the cinnamaldehyde-loaded self-microemulsion system not only has the advantages of simple preparation method, high drug loading, easy dispersion, good stability and easy large-scale production, but also has significant efficacy after cinnamaldehyde is used in combination with other Chinese medicine components, improves the drugability of the drug, has an improvement effect on liver fibrosis and has a significant therapeutic effect on non-alcoholic fatty hepatitis, and has certain application and development value. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The results of the anti-liver fibrosis efficacy of each group of mice are shown in the figure. From left to right, each column corresponds to the Sham group, BDL group, CIN group, DHI group, CR group, DR group, and CDR group; from top to bottom, each row corresponds to hematoxylin-eosin staining, picrosirius red staining, and Masson staining; Sham is the sham operation group; BDL is the liver fibrosis model group; CIN and DHI are cinnamaldehyde API and dihydrotanshinone I API, respectively; CR, DR, and CDR are cinnamaldehyde nanoemulsion, dihydrotanshinone I self-microemulsion, and cinnamaldehyde drug-loaded self-microemulsion, respectively.

[0017] Figure 2 The results of the efficacy of each group of mice against non-alcoholic fatty liver disease (NASH) are shown in the figure. From left to right, each column corresponds to the Control group, MCD group, CIN group, DHI group, MIX group, CR group, DR group, and CDR group; Control is the control group; MCD is the non-alcoholic fatty liver disease model group; CIN, DHI, and MIX are cinnamaldehyde API, dihydrotanshinone I, and the physical mixture of cinnamaldehyde API + dihydrotanshinone I, respectively; CR, DR, and CDR are cinnamaldehyde nanoemulsion, dihydrotanshinone I self-microemulsion, and cinnamaldehyde drug-loaded self-microemulsion, respectively. DETAILED DESCRIPTION

[0018] The present invention provides a cinnamaldehyde drug-carrying self-microemulsion, comprising the following raw materials in parts by weight: 0.5-5.5 parts of cinnamaldehyde, 0-5.5 parts of oil phase, 0.5-5.5 parts of emulsifier and 0.01-0.045 parts of poorly soluble drugs; the mass ratio of the cinnamaldehyde to the emulsifier is 1:0.9-3.5; preferably, the cinnamaldehyde drug-carrying self-microemulsion comprises the following raw materials in parts by weight: 0.8-4.5 parts of cinnamaldehyde, 0-4.5 parts of oil phase, 0.8-4.5 parts of emulsifier and 0.01-0.035 parts of drugs; the mass ratio of the cinnamaldehyde to the emulsifier is 1:0.9-1.1. When the oil phase of the present invention is not 0, the mass ratio of the oil phase to the emulsifier is 1:0.9-3.5, preferably 1:1-3.

[0019] In the present invention, the oil phase is preferably a carbon chain triglyceride; the emulsifier includes one or more of castor oil polyoxyethylene ether and polyoxyethylene hydrogenated castor oil, preferably castor oil polyoxyethylene ether or a combination of castor oil polyoxyethylene ether + polyoxyethylene hydrogenated castor oil. When castor oil polyoxyethylene ether + polyoxyethylene hydrogenated castor oil are combined, the mass ratio of castor oil polyoxyethylene ether and polyoxyethylene hydrogenated castor oil is preferably 1:0.5-2, and further preferably 1:1; the drug includes one or more of dihydrotanshinone I, dehydrocostus lactone, and costusinolone, preferably dihydrotanshinone I, dehydrocostus lactone or costusinolone.

[0020] In the present invention, as an embodiment, the cinnamaldehyde drug-loaded self-microemulsion comprises the following raw materials by weight: 1 part of cinnamaldehyde, 1 part of emulsifier and 0.01-0.035 parts of poorly soluble drugs. In this embodiment, the cinnamaldehyde is both a drug component and an oil phase. When the cinnamaldehyde of the present invention is an oil phase, it acts as an oil phase and an effective ingredient of the emulsifier.

[0021] In the present invention, as another embodiment, the cinnamaldehyde drug-loaded self-microemulsion comprises the following raw materials by weight: 1 part of cinnamaldehyde, 1 part of oil phase, 1 part of emulsifier and 0.01-0.035 parts of poorly soluble drugs. In this embodiment, cinnamaldehyde is used as the first raw material and also has the function of oil phase. The poorly soluble drug of the present invention is preferably dihydrotanshinone I, dehydrocostus lactone or costus alcohol lactone as the second raw material. The self-microemulsion prepared by this embodiment of the present invention is called cinnamaldehyde composite self-microemulsion.

[0022] The present invention provides a method for preparing the cinnamaldehyde drug-carrying self-microemulsion, comprising the following steps: uniformly mixing the weight portions of cinnamaldehyde, an oil phase, an emulsifier, and a poorly soluble drug to obtain the cinnamaldehyde drug-carrying self-microemulsion. The mixing speed of the present invention is 550-650rpm, preferably 580-630rpm, and more preferably 600rpm.

[0023] The present invention also provides the use of the cinnamaldehyde drug-loaded self-microemulsion in preparing a product for improving liver fibrosis. The liver fibrosis described in the present invention includes but is not limited to mid- and late-stage liver fibrosis.

[0024] The present invention also provides the use of the cinnamaldehyde drug-loaded self-microemulsion in preparing a product for treating fatty hepatitis. The fatty hepatitis described in the present invention includes but is not limited to early non-alcoholic fatty hepatitis.

[0025] In the present invention, unless otherwise specified, all raw materials are commercially available products well known to those skilled in the art.

[0026] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Example 1 A method for preparing a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 10mg of dihydrotanshinone I, and mix them evenly at a rotation speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0028] Example 2 A method for preparing a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1.2g of castor oil polyoxyethylene ether EL and 10mg of dihydrotanshinone I, and mix them evenly at a rotation speed of 600rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0029] Example 3 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of a mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH (the mass ratio of EL to RH is 1:1), and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600 rpm to obtain a cinnamaldehyde drug-loaded self-microemulsion.

[0030] Example 4 A method for preparing a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 20mg of dihydrotanshinone I, and mix them evenly at a rotation speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0031] Example 5 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 25mg of dihydrotanshinone I, and mix them evenly at a rotation speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0032] Example 6 A method for preparing a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 30mg of dihydrotanshinone I, and mix them evenly at a rotation speed of 600rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0033] Example 7 A method for preparing a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 35mg of dihydrotanshinone I, and mix them evenly at a rotation speed of 600rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0034] Example 8 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of medium-chain triglyceride, 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600 rpm to obtain a cinnamaldehyde drug-loaded self-microemulsion.

[0035] Example 9 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of medium-chain triglyceride, 1g of cinnamaldehyde, 2g of castor oil polyoxyethylene ether EL and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600rpm to obtain a cinnamaldehyde drug-loaded self-microemulsion.

[0036] Example 10 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of medium-chain triglyceride, 1g of cinnamaldehyde, 3g of castor oil polyoxyethylene ether EL and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0037] Embodiment 11 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of medium-chain triglyceride, 1g of cinnamaldehyde, 1g of a mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH (the mass ratio of EL to RH is 1:1), and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600 rpm to obtain a cinnamaldehyde drug-loaded self-microemulsion.

[0038] Example 12 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of medium-chain triglyceride, 1g of cinnamaldehyde, 2g of a mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH (the mass ratio of EL to RH is 1:1), and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600 rpm to obtain a cinnamaldehyde drug-loaded self-microemulsion.

[0039] Embodiment 13 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of medium-chain triglyceride, 1g of cinnamaldehyde, 3g of a mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH (the mass ratio of EL to RH is 1:1), and 10mg of dihydrotanshinone I, mix them evenly at a speed of 600 rpm to obtain a cinnamaldehyde drug-loaded self-microemulsion.

[0040] Embodiment 14 A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 10mg of dehydrocostus lactone, mix them evenly at a speed of 600rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0041] Embodiment 15 A method for preparing a cinnamaldehyde drug-loaded self-microemulsion is as follows: weigh 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 10mg of costusin lactone, and mix them evenly at a rotation speed of 600rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0042] Comparative Example 1 The difference from Example 1 is that castor oil polyoxyethylene ether EL is replaced by Tween 80, and the other steps are the same.

[0043] Comparative Example 2 The difference from Example 1 is that the castor oil polyoxyethylene ether EL is replaced by polyoxyethylene hydrogenated castor oil RH, and the other steps are the same.

[0044] Comparative Example 3 The difference from Example 1 is that the amount of castor oil polyoxyethylene ether EL is replaced with 0.4 g, and the other steps are the same.

[0045] Comparative Example 4 The difference from Example 1 is that the amount of castor oil polyoxyethylene ether EL is replaced with 0.6 g, and the other steps are the same.

[0046] Comparative Example 5 The difference from Example 1 is that the amount of castor oil polyoxyethylene ether EL is replaced with 0.8 g, and the other steps are the same.

[0047] Comparative Example 6 The difference from Example 3 is that the amount of the mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH is replaced with 0.4 g, and the remaining steps are the same.

[0048] Comparative Example 7 The difference from Example 3 is that the amount of the mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH is replaced with 0.6 g, and the remaining steps are the same.

[0049] Comparative Example 8 The difference from Example 3 is that the amount of the mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH is replaced with 0.8 g, and the remaining steps are the same.

[0050] Comparative Example 9 The difference from Example 3 is that the amount of the mixture of castor oil polyoxyethylene ether EL and polyoxyethylene hydrogenated castor oil RH is replaced with 1.2 g, and the remaining steps are the same.

[0051] Comparative Example 10 The difference from Example 1 is that the dosage of dihydrotanshinone I is replaced with 5 mg, and the remaining steps are the same.

[0052] Comparative Example 11 The difference from Example 1 is that the amount of dihydrotanshinone I is replaced with 40 mg, and the other steps are the same.

[0053] Comparative Example 12 The difference from Example 1 is that the amount of dihydrotanshinone I is replaced with 60 mg, and the other steps are the same.

[0054] Comparative Example 13 The difference from Example 1 is that the amount of dihydrotanshinone I is replaced with 80 mg, and the remaining steps are the same.

[0055] Comparative Example 14 The difference from Example 8 is that castor oil polyoxyethylene ether EL is replaced by Tween 80, and the other steps are the same.

[0056] Comparative Example 15 The difference from Comparative Example 14 is that the amount of Tween 80 is replaced with 2 g, and the other steps are the same.

[0057] Comparative Example 16 The difference from Comparative Example 14 is that the amount of Tween 80 is replaced with 3 g, and the other steps are the same.

[0058] Comparative Example 17 The difference from Example 8 is that the medium-chain triglyceride is replaced by olive oil, and the other steps are the same.

[0059] Comparative Example 18 The difference from Example 8 is that the medium-chain triglyceride is replaced by soybean oil, and the other steps are the same.

[0060] Comparative Example 19 The difference from Example 8 is that the medium-chain triglyceride is replaced by isopropyl palmitate, and the other steps are the same.

[0061] Comparative Example 20 The difference from Example 8 is that the medium-chain triglyceride is replaced by octanoic acid glyceride, and the remaining steps are the same.

[0062] Comparative Example 21 The difference from Example 8 is that the medium-chain triglyceride is replaced by isopropyl myristate, and the remaining steps are the same.

[0063] Comparative Example 22 The difference from Example 7 is that 1 g of castor oil polyoxyethylene ether EL is replaced with 0.5 g of castor oil polyoxyethylene ether EL, and the other steps are the same.

[0064] Experimental Example 1 1. Particle size, PDI and appearance of cinnamaldehyde drug-loaded self-microemulsions prepared in Examples 1-3, Comparative Examples 1-9 and Comparative Examples 17-22 3mL of water was added dropwise to the cinnamaldehyde drug-loaded self-microemulsion prepared in each group above, and the speed was maintained at 600rpm during the dropping process, and a uniform dispersion system was spontaneously formed, namely, cinnamaldehyde drug-loaded nanoemulsion. The cinnamaldehyde drug-loaded nanoemulsion was placed at 37°C for 0h, 8h, 12h, and 24h, and its appearance properties were observed, and the average particle size and PDI were measured using a Malvern laser particle size analyzer to simulate the stability of the cinnamaldehyde drug-loaded self-microemulsion entering the body and being emulsified into nanoemulsion through gastrointestinal fluid. The results are shown in Table 1.

[0065] 2. Study on the storage stability of the cinnamaldehyde drug-loaded self-microemulsions prepared in Examples 1-3, Comparative Examples 3-9 and Comparative Examples 17-21 After each group of cinnamaldehyde drug-loaded self-microemulsions were placed at room temperature, their appearance and properties were observed, and their particle sizes and PDI were measured to investigate their storage stability. The results are shown in Table 2.

[0066] The results in Tables 1 and 2 show that when cinnamaldehyde is used as the oil phase and Tween 80 is used as the emulsifier, it is difficult to form an emulsion with uniform particle size, the PDI is large, and stratification and drug precipitation occur after being placed; when cinnamaldehyde is used as the oil phase and the emulsifier is polyoxyethylene hydrogenated castor oil RH, self-microemulsion and nanoemulsion with uniform particle size can be formed, but stratification occurs after being placed at 37°C for 12 hours, and the stability is poor at body temperature; when the emulsifier is castor oil polyoxyethylene ether EL or a mixture of RH+EL, self-microemulsion and nanoemulsion with uniform particle size can be formed, and the placement stability is good. When the ratio of oil phase to emulsion is less than 1:1, stratification is more likely to occur after long-term placement.

[0067] When the oil phase was replaced with soybean oil, olive oil, isopropyl palmitate, caprylic decanoate and isopropyl myristate, the drug was easy to precipitate and stratify in the microemulsion, and it was difficult to form stable and uniform nanoparticles within 24 hours at 37°C.

[0068] Table 1 Results of selection of emulsifier and oil phase for cinnamaldehyde drug-loaded self-microemulsifier

[0069] Note: - represents not measured.

[0070] Table 2 The results of the storage stability of each group of cinnamaldehyde drug-loaded self-microemulsions

[0071] Note: - represents not measured.

[0072] Experimental Example 2 3mL of water was added dropwise to each group of cinnamaldehyde drug-loaded self-microemulsions (cinnamaldehyde drug-loaded self-microemulsions prepared in Example 1, Examples 4-7 and Comparative Examples 10-13), and the speed was maintained at 600rpm during the dropwise addition process to spontaneously form a uniform dispersion system, i.e., cinnamaldehyde drug-loaded nanoemulsion. The cinnamaldehyde drug-loaded nanoemulsion was placed at 37°C for 0h, 8h, 12h, and 24h, and its appearance properties were observed, and the particle size and PDI were measured to simulate the stability of the cinnamaldehyde drug-loaded self-microemulsion entering the body and being emulsified into nanoemulsions through gastrointestinal fluid. The results are shown in Table 3.

[0073] The optimal drug loading of dihydrotanshinone I can be obtained from Table 3: when the oil phase is cinnamaldehyde, castor oil polyoxyethylene ether EL is the emulsifier, and the addition amount of dihydrotanshinone I is ≥40 mg, the drug precipitates within 3 days and a uniform emulsion cannot be formed; when the addition amount of dihydrotanshinone I is ≤35 mg, cinnamaldehyde-loaded self-microemulsions and nanoemulsions with uniform particle size can be formed, and the particle size and PDI do not change significantly at 37°C and room temperature. When the amount of cinnamaldehyde is 1 g and the amount of dihydrotanshinone I is 35 mg, the maximum drug loading amount of cinnamaldehyde-loaded dihydrotanshinone I self-microemulsion can be obtained.

[0074] Table 3 Screening results of maximum drug loading capacity of self-microemulsion

[0075] Note: - represents not measured.

[0076] Experimental Example 3 The storage stability of the cinnamaldehyde drug-loaded self-microemulsions prepared in Examples 8-13 and Comparative Examples 14-16 was investigated.

[0077] After each group of cinnamaldehyde drug-loaded self-microemulsions were placed at room temperature away from light, their appearance and properties were observed, and their particle sizes and PDI were measured to investigate their storage stability. The results are shown in Table 4.

[0078] From the results in Table 4, it can be seen that when the emulsifier is Tween 80, the self-emulsifying system is easy to stratify; when the emulsifier is castor oil polyoxyethylene ether EL or a mixture of RH+EL, a stable self-microemulsion can be formed after contacting water, and the self-microemulsion system has good storage stability. When the mass ratio of the emulsifier to the oil phase is greater than or equal to 50%, it is easy to form an emulsion with relatively uniform particle size when contacting water; on the contrary, when the mass ratio of the emulsifier to the oil phase is less than 50%, it is difficult to emulsify and form an emulsion with uniform particle size when contacting water. Therefore, a self-microemulsifying system with a ratio of the emulsifier to the oil phase greater than or equal to 50% is preferred.

[0079] Table 4 Results of room temperature placement of each group of cinnamaldehyde drug-loaded self-microemulsions

[0080] Note: - means not tested Experimental Example 4 The stability of the cinnamaldehyde drug-loaded self-microemulsions prepared in Examples 7, 14 and 15 was investigated.

[0081] The cinnamaldehyde drug-loaded self-microemulsions prepared in the above groups were placed in vials and stored at 4°C in the dark. The particle size, PDI and cinnamaldehyde and drug content were detected by high performance liquid chromatography after 0, 3, 7, 15, 30, 45 and 60 days, respectively. The results are shown in Table 5. It can be seen from the results in the table that the particle size and PDI of the cinnamaldehyde drug-loaded self-microemulsion did not change significantly within 60 days, and the remaining percentage of cinnamaldehyde and drug content did not change significantly.

[0082] Table 5 The results of the storage stability of each group of cinnamaldehyde drug-loaded self-microemulsions

[0083] Experimental Example 5: Drug efficacy experiment on rat liver fibrosis Experimental animals: SD male rats, weighing 180-200 g.

[0084] Construction of animal model of cholestatic liver fibrosis (BDL): 35 experimental animals were fasted for 12 hours before surgery; after anesthesia with isoflurane, the abdomen was opened under sterile conditions, the liver edge was raised, the duodenum was pulled open, the common bile duct was separated by 2-3 cm, and two ligatures were made near the duodenum and near the liver hilum with No. 000 silk thread, the common bile duct was cut between the two ligatures, the liver was restored to its original position and the incision was sutured, and after the animals woke up from anesthesia, they ate normally and drank water freely.

[0085] Model group (5 animals): 5 of the above BDL model animals were selected as the BDL model group (abbreviated as BDL), and physiological saline was given by gavage once a day starting from the second day after surgery.

[0086] Sham operation group (5 animals): 5 experimental animals were selected as the sham operation group (abbreviated as Sham). They were fasted for 12 hours before the operation. After anesthesia with isoflurane, the abdomen was opened under sterile conditions, and then the incision was sutured. After the animals woke up from anesthesia, they ate normally and drank water freely. Starting from the second day after the operation, they were gavaged with normal saline once a day.

[0087] API group (10 animals): 10 of the above BDL model animals were selected as the API group (referred to as CIN and DHI), with 5 animals in each group. Starting from the second day after surgery, cinnamaldehyde (referred to as CIN) and dihydrotanshinone I (referred to as DHI) were given by gavage once a day. Cinnamaldehyde was evenly dispersed with olive oil; dihydrotanshinone I was evenly dispersed with CMC-Na solution. The single dose was about 82 mg / kg / day for cinnamaldehyde and about 2.9 mg / kg / day for dihydrotanshinone I.

[0088] Emulsion group (15 animals): 15 of the above BDL modeling animals were taken as the emulsion group (abbreviated as CR, DR and CDR). Starting from the second day after surgery, the cinnamaldehyde self-microemulsion (abbreviated as CDR) prepared in Example 7, the cinnamaldehyde self-microemulsion (abbreviated as CR) prepared under the same prescription as Example 7, or the dihydrotanshinone I self-microemulsion (abbreviated as DR) was gavaged once a day. The single dose was about 82 mg / kg / day for cinnamaldehyde; and about 2.9 mg / kg / day for dihydrotanshinone I. Among them, the preparation method of dihydrotanshinone I self-microemulsion (abbreviated as DR) is as follows: DHI 0.14%, medium-chain triglyceride 20.26%, emulsifier polyoxyethylene hydrogenated castor oil (RH) 31.36% and co-emulsifier PEG-400 48.24%.

[0089] After 14 days of drug administration (or saline), the above groups fasted for 12 hours, and then collected blood, liver tissue and other samples. Serum was taken for serum biochemical index detection. The results of serum biochemical index detection are shown in Table 6. Compared with the Sham group, the levels of ALT, AST, ALP, TBA and TBiLi in the BDL model group were significantly increased. Compared with the BDL model group, the CDR group can significantly reduce the levels of AST, TBA and TBiLi in rat serum, indicating that cinnamaldehyde-loaded self-microemulsion can significantly improve the liver function level of BDL rats, and the effect is better than that of the raw material drug and single drug-loaded emulsion.

[0090] Table 6 Results of anti-liver fibrosis efficacy indexes

[0091] Note: *** indicates significant difference compared with the sham operation group (Sham), P<0.001; ** indicates significant difference compared with the sham operation group (Sham), P<0.01; # indicates significant difference compared with the model group (BDL), P<0.05.

[0092] Paraffin sections were prepared from liver tissue samples of each group and stained with hematoxylin-eosin (HE), Sirius red and Masson respectively. Figure 1 As shown. The results showed that compared with the Sham group, the BDL model group showed obvious hepatocyte necrosis, bile duct hyperplasia, collagen deposition, inflammatory infiltration and fibrosis in the liver, indicating that the model was successfully established. The CIN and DHI groups had no significant improvement in bile duct hyperplasia, hepatocyte necrosis and collagen deposition in the liver, while CR, DR and CDR could reduce bile duct hyperplasia, hepatocyte necrosis and collagen deposition. However, there was still some inflammatory infiltration and collagen deposition in the CDR group, indicating that cinnamaldehyde-loaded self-microemulsion has a certain improvement effect on the middle and late stages of liver fibrosis, and can delay the process of liver fibrosis, but cannot reverse it.

[0093] Experimental Example 6: Efficacy of Nonalcoholic Steatohepatitis (NASH) Experimental animals: C57BL / 6 male mice, weighing 25-28 g.

[0094] Nonalcoholic steatohepatitis (MCD) animal model: A total of 64 experimental animals were randomly divided into 8 groups, with 8 animals in each group. All groups were given an adaptive diet for the first 3 days, and then the MCD diet was given for 7 weeks starting from the fourth day to establish the NASH model. Continuous drug administration began from the 5th week.

[0095] Model group (8 animals): 8 of the above MCD model animals were selected as the MCD model group (MCD for short). Starting from the 5th week, they were given normal saline by gavage once a day.

[0096] Control group (8 animals): 8 experimental animals were selected as the control group (abbreviated as Control) and given a normal diet for 7 consecutive weeks. Starting from the 5th week, they were given normal saline by gavage once a day.

[0097] API group (24 animals): 24 of the above MCD model animals were selected as the API group, with 8 animals in each group. Starting from the 5th week, cinnamaldehyde API (CIN), dihydrotanshinone I API (DHI), and the physical mixture (MIX) of cinnamaldehyde and dihydrotanshinone I were given by gavage once a day. Cinnamaldehyde was evenly dispersed with olive oil; dihydrotanshinone I was evenly dispersed with CMC-Na solution. The single dose was about 82 mg / kg / day for cinnamaldehyde; and about 2.9 mg / kg / day for dihydrotanshinone I.

[0098] Emulsion group (24 animals): 24 of the above MCD model animals were taken as the emulsion group. Starting from the 5th week, the cinnamaldehyde drug-loaded self-microemulsion (CDR) prepared in Example 7, and the cinnamaldehyde nanoemulsion (CR) and dihydrotanshinone I self-microemulsion (DR) prepared in step 1 of Experimental Example 7 under the same prescription were administered by gavage once a day. The single dose of cinnamaldehyde was about 82 mg / kg / day; dihydrotanshinone I was about 2.9 mg / kg / day.

[0099] After 7 weeks, fast for 12 hours, and then collect blood, liver tissue and other samples. Serum was collected for serum biochemical index detection. The results of serum biochemical index detection are shown in Tables 7 and 8. Compared with the Control group, the levels of ALT, AST, ALP and TBA in the MCD model group were significantly increased. Compared with the MCD model group, the CDR administration group reversed the serum indicators of MCD mice to near normal levels, while the raw material drug group and the single drug-loaded emulsion group could only improve 1-3 of the indicators. In summary, the results show that cinnamaldehyde-loaded self-microemulsion can significantly improve the liver function level of MCD mice, and the effect is better than the raw material drug and the single drug-loaded emulsion and their physical mixture.

[0100] Table 7 Results of efficacy indexes against non-alcoholic fatty liver disease 1

[0101] Table 8 Anti-nonalcoholic fatty liver disease efficacy index results 2

[0102]

[0103] Note: In Table 7 and Table 8, *** indicates that compared with the Control group, there was a significant difference, P<0.001; ** indicates that compared with the Control group, there was a significant difference, P<0.01; * indicates that compared with the Control group, there was a significant difference, P<0.05; ### indicates that compared with the MCD group, there was a significant difference, P<0.001; ## indicates that compared with the MCD group, there was a significant difference, P<0.01; # indicates that compared with the MCD group, there was a significant difference, P<0.05.

[0104] The liver tissue samples of each group were stained with hematoxylin-eosin (HE) and oil red, and the staining results were shown in Figure 2 As shown. The results showed that compared with the Control group, a large amount of fat vacuoles accumulation, lipid degeneration and inflammatory infiltration appeared in the liver of the MCD model group, indicating that the non-alcoholic fatty liver disease model was successfully established. Compared with the MCD group, the CR and DR groups also significantly reduced liver fat accumulation and inflammatory infiltration, while there were no obvious fat vacuoles in the CDR group, reversing non-alcoholic fatty liver disease. However, there was no significant improvement in liver fat accumulation and inflammatory infiltration in the CIN, DHI and CIN and DHI physical mixture groups. In summary, the results show that cinnamaldehyde-loaded self-microemulsion has a significant therapeutic effect on early non-alcoholic fatty liver disease, can promote lipolysis, reduce the occurrence of inflammation, and reverse non-alcoholic fatty liver disease.

[0105] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A cinnamaldehyde drug-loaded self-microemulsion, characterized in that: The invention comprises the following raw materials in parts by weight: 0.5-5.5 parts of cinnamaldehyde, 0-5.5 parts of oil phase, 0.5-5.5 parts of emulsifier and 0.01-0.045 parts of poorly soluble drugs; the mass ratio of cinnamaldehyde to emulsifier is 1:0.5-3.

5.

2. A cinnamaldehyde drug-loaded self-microemulsion, characterized in that: The invention comprises the following raw materials in parts by weight: 0.8-4.5 parts of cinnamaldehyde, 0-4.5 parts of oil phase, 0.8-4.5 parts of emulsifier and 0.01-0.035 parts of poorly soluble drugs; the mass ratio of cinnamaldehyde to emulsifier is 1:0.9-1.

1.

3. The cinnamaldehyde drug-loaded self-microemulsion according to claim 1 or 2, characterized in that: The mass ratio of the oil phase to the emulsifier is 1:0.9-3.

5.

4. The cinnamaldehyde drug-loaded self-microemulsion according to claim 1 or 2, characterized in that: The oil phase includes medium-chain triglycerides.

5. The cinnamaldehyde drug-loaded self-microemulsion according to claim 1 or 2, characterized in that: The emulsifier includes one or more of castor oil polyoxyethylene ether and polyoxyethylene hydrogenated castor oil.

6. The cinnamaldehyde drug-loaded self-microemulsion according to claim 1 or 2, characterized in that: The poorly soluble drugs include one or more of dihydrotanshinone I, dehydrocostus lactone and costusinolone.

7. The method for preparing the cinnamaldehyde drug-loaded self-microemulsion according to any one of claims 1 to 6, characterized in that: The method comprises the following steps: uniformly mixing the weight portions of cinnamaldehyde, oil phase, emulsifier and poorly soluble drugs to obtain cinnamaldehyde drug-loaded self-microemulsion.

8. The preparation method according to claim 7, characterized in that: The mixing speed is 550-650 rpm.

9. Use of the cinnamaldehyde drug-loaded self-microemulsion according to any one of claims 1 to 6 or the cinnamaldehyde drug-loaded self-microemulsion obtained by the preparation method according to any one of claims 7 to 8 in preparing a product for improving liver fibrosis.

10. Use of the cinnamaldehyde drug-loaded self-microemulsion according to any one of claims 1 to 6 or the cinnamaldehyde drug-loaded self-microemulsion obtained by the preparation method according to any one of claims 7 to 8 in preparing a product for treating fatty hepatitis.

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