A cinnamaldehyde drug-loaded self-microemulsion and its preparation method and application

Abstract

The present invention provides a cinnamaldehyde drug-loaded self-microemulsion and its preparation method and application, belonging to the field of biopharmaceutical technology. The cinnamaldehyde drug-loaded self-microemulsion of the present invention includes 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 present invention adopts a self-microemulsion drug delivery system to improve the efficacy of cinnamaldehyde and cinnamaldehyde and other traditional Chinese medicine ingredients after being used together, with high drug loading, easy dispersion and high stability, and cinnamaldehyde and other traditional Chinese medicine ingredients have significant drug efficacy after being used together, have an improvement effect on liver fibrosis and have a significant therapeutic effect on non-alcoholic fatty liver disease, and have certain application and development value.

Description

Technical Field

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

[0002] Biochemical tests for patients with non-alcoholic fatty liver disease (NAFLD) reveal elevated triglycerides, elevated LDL cholesterol, and decreased HDL cholesterol. These symptoms are often associated with features of the metabolic syndrome, such as obesity, dyslipidemia, type 2 diabetes, and hypertension. Hepatic steatosis and lipid accumulation are the earliest stages of NAFLD and are typically asymptomatic. Appropriate dietary or lifestyle changes can revert the affected liver to a healthy state. Subsequently, lipotoxicity, oxidative stress, and endoplasmic reticulum stress damage hepatocytes, triggering extensive inflammation, and fatty liver transforms into non-alcoholic steatohepatitis (NASH). Without intervention, ongoing liver damage and inflammation further activate hepatic stellate cells, secreting collagen and leading to liver fibrosis. Simple steatosis and NASH are reversible stages of NAFLD, while advanced stages, such as cirrhosis and even liver cancer, can only be treated with liver transplantation and are irreversible. However, there are currently few therapeutic agents available for each stage of NAFLD. Therefore, the development of novel anti-NAFLD drugs is urgently needed, especially for early and middle stages of fatty liver, NASH, and liver fibrosis.

[0003] Cinnamaldehyde (CIN) is an aldehyde-like 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 pale yellow oily liquid at room temperature and has antibacterial, lipid-lowering, and antioxidant properties. Costunolide and dehydrocostuslactone are substances extracted from the roots of Costus vulgaris (Asteraceae) and have smooth muscle relaxant and antispasmodic effects. Dihydrotanshinone I is an active substance from the traditional Chinese medicine Salvia miltiorrhiza, with antibacterial, platelet aggregation inhibitory, antioxidant, and coronary vasodilatory activities. However, the clinical application of cinnamaldehyde, costunolide, dehydrocostuslactone, and dihydrotanshinone I is severely limited by their low solubility, poor lipid solubility, poor gastrointestinal absorption, and rapid metabolism. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide a cinnamaldehyde-loaded self-microemulsion and its preparation method and application. The cinnamaldehyde-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 capacity, easy dispersion and good stability. It has an improvement effect on liver fibrosis and a significant therapeutic effect on non-alcoholic fatty liver disease.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0006] The invention provides a cinnamaldehyde drug-loaded 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.

[0007] 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.

[0008] Preferably, the mass ratio of the oil phase to the emulsifier is 1:0.9-3.5.

[0009] Preferably, the oil phase comprises carbon chain triglycerides.

[0010] Preferably, the emulsifier includes one or more of castor oil polyoxyethylene ether and polyoxyethylene hydrogenated castor oil.

[0011] Preferably, the poorly soluble drug includes one or more of dihydrotanshinone I, dehydrocostus lactone, and costus lactone.

[0012] The invention provides a preparation method of the cinnamaldehyde drug-loaded self-microemulsion, comprising the following steps: uniformly mixing the cinnamaldehyde, oil phase, emulsifier and drug in parts by weight to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0013] Preferably, the mixing speed is 550-650 rpm.

[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 improving liver fibrosis.

[0015] 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 treating fatty liver disease.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention utilizes a self-microemulsifying drug delivery system to enhance the efficacy of cinnamaldehyde and its combination with other traditional Chinese medicines. Furthermore, the cinnamaldehyde in the cinnamaldehyde-loaded self-microemulsion not only serves as a pharmaceutical ingredient but also as the oil phase of the self-microemulsion. Compared to a single nanoemulsion system, the cinnamaldehyde-loaded self-microemulsion system not only has the advantages of a simple preparation method, high drug loading, ease of dispersion, good stability, and ease of large-scale production, but also exhibits significant efficacy when cinnamaldehyde is combined with other traditional Chinese medicine ingredients, thereby enhancing the drugability of the drug, improving liver fibrosis symptoms, and having a significant therapeutic effect on non-alcoholic steatohepatitis, thus possessing considerable application and development value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The graph shows the anti-hepatic fibrosis efficacy results for each group of mice. 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 represents the sham operation group; BDL represents the liver fibrosis model group; CIN and DHI represent the cinnamaldehyde API and dihydrotanshinone I API, respectively; CR, DR, and CDR represent the cinnamaldehyde nanoemulsion, dihydrotanshinone I self-microemulsion, and cinnamaldehyde-loaded self-microemulsion, respectively.

[0019] Figure 2 The graph shows the efficacy of the drug against non-alcoholic steatohepatitis (NASH) in each group of mice. From left to right, the columns correspond to the Control, MCD, CIN, DHI, MIX, CR, DR, and CDR groups, respectively. Control represents the control group; MCD represents the non-alcoholic steatohepatitis model group; CIN, DHI, and MIX represent the cinnamaldehyde API, dihydrotanshinone I, and a physical mixture of the cinnamaldehyde API and dihydrotanshinone I, respectively; CR, DR, and CDR represent the cinnamaldehyde nanoemulsion, dihydrotanshinone I self-microemulsion, and cinnamaldehyde-loaded self-microemulsion, respectively. DETAILED DESCRIPTION

[0020] The present invention provides a cinnamaldehyde drug-loaded 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-loaded 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.

[0021] 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, more preferably 1:1; the drug includes one or more of dihydrotanshinone I, dehydrocostus lactone, and costus lactone, preferably dihydrotanshinone I, dehydrocostus lactone or costus lactone.

[0022] In one embodiment of the present invention, the cinnamaldehyde drug-loaded self-microemulsion comprises the following raw materials by weight: 1 part cinnamaldehyde, 1 part emulsifier, and 0.01-0.035 parts of a poorly soluble drug. In this embodiment, the cinnamaldehyde serves as both the drug component and the oil phase. When the cinnamaldehyde serves as the oil phase, it acts as both the oil phase of the emulsifier and the active ingredient.

[0023] In another embodiment of the present invention, the cinnamaldehyde-loaded drug self-microemulsion comprises the following raw materials in parts by weight: 1 part cinnamaldehyde, 1 part oil phase, 1 part emulsifier, and 0.01-0.035 parts of a poorly soluble drug. In this embodiment, cinnamaldehyde, as the first API, also functions as an oil phase. The poorly soluble drug of the present invention is preferably dihydrotanshinone I, dehydrocostus lactone, or costunolide, as the second raw material. The self-microemulsion prepared by this embodiment of the present invention is referred to as a cinnamaldehyde composite self-microemulsion.

[0024] The present invention provides a method for preparing the cinnamaldehyde drug-loaded self-microemulsion, comprising the steps of: uniformly mixing parts by weight of cinnamaldehyde, an oil phase, an emulsifier, and a poorly soluble drug to obtain the cinnamaldehyde drug-loaded self-microemulsion. The mixing speed of the present invention is 550-650 rpm, preferably 580-630 rpm, and more preferably 600 rpm.

[0025] 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- to late-stage liver fibrosis.

[0026] The present invention also provides the use of the cinnamaldehyde drug-loaded self-microemulsion in preparing a product for treating steatohepatitis. The steatohepatitis described in the present invention includes but is not limited to early non-alcoholic steatohepatitis.

[0027] In the present invention, unless otherwise specified, all raw materials are commercially available products well known to those skilled in the art.

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some 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 making creative efforts are within the scope of protection of the present invention.

[0029] Example 1

[0030] 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 dihydrotanshinone I, and mix them uniformly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0031] Example 2

[0032] A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: 1 g of cinnamaldehyde, 1.2 g of castor oil polyoxyethylene ether EL and 10 mg of dihydrotanshinone I are weighed and mixed uniformly at a rotation speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0033] Example 3

[0034] A method for preparing a cinnamaldehyde-loaded drug self-microemulsion is as follows: weigh 1 g of cinnamaldehyde, 1 g 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 10 mg of dihydrotanshinone I, and mix them uniformly at a rotation speed of 600 rpm to obtain the cinnamaldehyde-loaded drug self-microemulsion.

[0035] Example 4

[0036] 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 20mg of dihydrotanshinone I, and mix them uniformly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0037] Example 5

[0038] 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 uniformly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0039] Example 6

[0040] 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 30mg of dihydrotanshinone I, and mix them uniformly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0041] Example 7

[0042] 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 35mg of dihydrotanshinone I, and mix them uniformly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0043] Example 8

[0044] A preparation method of a cinnamaldehyde-loaded self-microemulsion is as follows: 1 g of medium-chain triglyceride, 1 g of cinnamaldehyde, 1 g of castor oil polyoxyethylene ether EL and 10 mg of dihydrotanshinone I are weighed and mixed uniformly at a speed of 600 rpm to obtain the cinnamaldehyde-loaded self-microemulsion.

[0045] Example 9

[0046] A preparation method of a cinnamaldehyde-loaded self-microemulsion is as follows: 1 g of medium-chain triglyceride, 1 g of cinnamaldehyde, 2 g of castor oil polyoxyethylene ether EL and 10 mg of dihydrotanshinone I are weighed and mixed uniformly at a rotation speed of 600 rpm to obtain the cinnamaldehyde-loaded self-microemulsion.

[0047] Example 10

[0048] A preparation method of a cinnamaldehyde-loaded self-microemulsion is as follows: 1 g of medium-chain triglyceride, 1 g of cinnamaldehyde, 3 g of castor oil polyoxyethylene ether EL and 10 mg of dihydrotanshinone I are weighed and mixed uniformly at a rotation speed of 600 rpm to obtain the cinnamaldehyde-loaded self-microemulsion.

[0049] Example 11

[0050] A method for preparing a cinnamaldehyde-loaded self-microemulsion is as follows: weigh 1 g of medium-chain triglyceride, 1 g of cinnamaldehyde, 1 g 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 10 mg of dihydrotanshinone I, and mix them uniformly at a rotation speed of 600 rpm to obtain a cinnamaldehyde-loaded self-microemulsion.

[0051] Example 12

[0052] A method for preparing a cinnamaldehyde-loaded self-microemulsion is as follows: weigh 1 g of medium-chain triglyceride, 1 g of cinnamaldehyde, 2 g 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 10 mg of dihydrotanshinone I, and mix them uniformly at a rotation speed of 600 rpm to obtain a cinnamaldehyde-loaded self-microemulsion.

[0053] Example 13

[0054] A method for preparing a cinnamaldehyde-loaded drug self-microemulsion is as follows: weigh 1 g of medium-chain triglyceride, 1 g of cinnamaldehyde, 3 g 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 10 mg of dihydrotanshinone I, and mix them uniformly at a rotation speed of 600 rpm to obtain a cinnamaldehyde-loaded drug self-microemulsion.

[0055] Example 14

[0056] A preparation method of a cinnamaldehyde drug-loaded self-microemulsion is as follows: 1g of cinnamaldehyde, 1g of castor oil polyoxyethylene ether EL and 10mg of dehydrocostus lactone are weighed and mixed uniformly at a rotation speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0057] Example 15

[0058] 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 costus lactone, and mix them uniformly at a speed of 600 rpm to obtain the cinnamaldehyde drug-loaded self-microemulsion.

[0059] Comparative Example 1

[0060] The difference from Example 1 is that castor oil polyoxyethylene ether EL is replaced by Tween 80, and the other steps are the same.

[0061] Comparative Example 2

[0062] The difference from Example 1 is that castor oil polyoxyethylene ether EL is replaced by polyoxyethylene hydrogenated castor oil RH, and the remaining steps are the same.

[0063] Comparative Example 3

[0064] 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.

[0065] Comparative Example 4

[0066] 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.

[0067] Comparative Example 5

[0068] 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.

[0069] Comparative Example 6

[0070] 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.

[0071] Comparative Example 7

[0072] 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.

[0073] Comparative Example 8

[0074] 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.

[0075] Comparative Example 9

[0076] 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.

[0077] Comparative Example 10

[0078] The difference from Example 1 is that the dosage of dihydrotanshinone I is replaced with 5 mg, and the remaining steps are the same.

[0079] Comparative Example 11

[0080] The difference from Example 1 is that the dosage of dihydrotanshinone I is replaced with 40 mg, and the remaining steps are the same.

[0081] Comparative Example 12

[0082] The difference from Example 1 is that the dosage of dihydrotanshinone I is replaced with 60 mg, and the remaining steps are the same.

[0083] Comparative Example 13

[0084] The difference from Example 1 is that the amount of dihydrotanshinone I is replaced with 80 mg, and the remaining steps are the same.

[0085] Comparative Example 14

[0086] The difference from Example 8 is that castor oil polyoxyethylene ether EL is replaced by Tween 80, and the other steps are the same.

[0087] Comparative Example 15

[0088] The difference from Comparative Example 14 is that the amount of Tween 80 is replaced with 2 g, and the remaining steps are the same.

[0089] Comparative Example 16

[0090] The difference from Comparative Example 14 is that the amount of Tween 80 is replaced with 3 g, and the remaining steps are the same.

[0091] Comparative Example 17

[0092] The difference from Example 8 is that the medium-chain triglyceride is replaced by olive oil, and the remaining steps are the same.

[0093] Comparative Example 18

[0094] The difference from Example 8 is that the medium-chain triglyceride is replaced by soybean oil, and the other steps are the same.

[0095] Comparative Example 19

[0096] The difference from Example 8 is that the medium-chain triglyceride is replaced by isopropyl palmitate, and the remaining steps are the same.

[0097] Comparative Example 20

[0098] The difference from Example 8 is that the medium-chain triglyceride is replaced by octanoic acid glyceride, and the remaining steps are the same.

[0099] Comparative Example 21

[0100] The difference from Example 8 is that the medium-chain triglyceride is replaced by isopropyl myristate, and the remaining steps are the same.

[0101] Comparative Example 22

[0102] 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.

[0103] Experimental Example 1

[0104] 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

[0105] 3 mL of water was added dropwise to each of the cinnamaldehyde-loaded self-microemulsions prepared above, maintaining a rotational speed of 600 rpm. A uniform dispersion, i.e., the cinnamaldehyde-loaded nanoemulsion, spontaneously formed. The cinnamaldehyde-loaded nanoemulsions were then incubated at 37°C for 0, 8, 12, and 24 hours. Their appearance and properties were observed, and their average particle size and particle size distribution (PDI) were measured using a Malvern laser particle size analyzer to simulate the in vivo stability of the cinnamaldehyde-loaded self-microemulsions upon intestinal emulsification into nanoemulsions. The results are shown in Table 1.

[0106] 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

[0107] After each group of cinnamaldehyde-loaded self-microemulsions were placed at room temperature, their appearance and properties were observed, and their particle size and PDI were measured to investigate their shelf stability. The results are shown in Table 2.

[0108] The results in Tables 1 and 2 show that when cinnamaldehyde was used as the oil phase and Tween 80 was used as the emulsifier, uniform emulsions were difficult to form, the PDI was large, and stratification and drug precipitation occurred after storage. When cinnamaldehyde was used as the oil phase and polyoxyethylene hydrogenated castor oil (RH) was used as the emulsifier, uniform self-microemulsions and nanoemulsions were formed, but stratification occurred after 12 hours at 37°C, indicating poor stability at in vivo temperatures. When polyoxyethylene castor oil ether (EL) or a mixture of RH and EL was used as the emulsifier, uniform self-microemulsions and nanoemulsions were formed, with good storage stability. When the oil phase to emulsion ratio was less than 1:1, stratification was more likely to occur after prolonged storage.

[0109] When the oil phase was replaced with soybean oil, olive oil, isopropyl palmitate, caprylic glyceride and isopropyl myristate, the drug was easily precipitated and stratified in the microemulsion, and it was difficult to form stable and uniform nanoparticles within 24 hours at 37°C.

[0110] Table 1 Results of selection of emulsifier and oil phase for cinnamaldehyde drug-loaded self-microemulsification

[0111]

[0112] Note: - represents not measured.

[0113] Table 2 Storage stability results of cinnamaldehyde drug-loaded self-microemulsions in each group

[0114]

[0115] Note: - represents not measured.

[0116] Experimental Example 2

[0117] 3 mL of water was added dropwise to each set of cinnamaldehyde-loaded SEMs (prepared in Example 1, Examples 4-7, and Comparative Examples 10-13). The rotational speed was maintained at 600 rpm during the addition process, resulting in the spontaneous formation of a uniform dispersion system, i.e., the cinnamaldehyde-loaded nanoemulsion. The cinnamaldehyde-loaded nanoemulsions were then incubated at 37°C for 0, 8, 12, and 24 hours. Their appearance and properties were observed, and their particle size and PDI were measured to simulate the stability of the cinnamaldehyde-loaded SEMs upon in vivo emulsification into nanoemulsions via gastrointestinal fluid. The results are shown in Table 3.

[0118] Table 3 shows the optimal drug loading of dihydrotanshinone I. When cinnamaldehyde was the oil phase, castor oil polyoxyethylene ether EL was the emulsifier, and the added amount of dihydrotanshinone I was ≥40 mg, drug precipitation occurred within 3 days, preventing the formation of a uniform emulsion. When the added amount of dihydrotanshinone I was ≤35 mg, uniform cinnamaldehyde-loaded self-microemulsions and nanoemulsions were formed, and the particle size and PDI did not change significantly at both 37°C and room temperature. The maximum drug loading of cinnamaldehyde-loaded dihydrotanshinone I self-microemulsions was achieved when the cinnamaldehyde dosage was 1 g and the dihydrotanshinone I dosage was 35 mg.

[0119] Table 3 Screening results of maximum drug loading capacity of self-microemulsion

[0120]

[0121] Note: - represents not measured.

[0122] Experimental Example 3

[0123] The storage stability of the cinnamaldehyde drug-loaded self-microemulsions prepared in Examples 8-13 and Comparative Examples 14-16 was investigated.

[0124] After each group of cinnamaldehyde-loaded self-microemulsions were placed at room temperature in the dark, their appearance and properties were observed, and their particle size and PDI were measured to investigate their shelf stability. The results are shown in Table 4.

[0125] The results in Table 4 show that when the emulsifier is Tween 80, the self-emulsifying system easily separates. However, when the emulsifier is castor oil polyoxyethylene ether EL or a mixture of RH and EL, stable self-microemulsions are formed upon contact with water, and the self-microemulsification system exhibits good shelf stability. When the mass ratio of emulsifier to oil phase is greater than or equal to 50%, an emulsion with relatively uniform particle size is easily formed upon contact with water. Conversely, when the mass ratio of emulsifier to oil phase is less than 50%, it is difficult to emulsify the emulsion with uniform particle size upon contact with water. Therefore, a self-microemulsification system with a mass ratio of emulsifier to oil phase greater than or equal to 50% is preferred.

[0126] Table 4 Results of room temperature storage of cinnamaldehyde loaded self-microemulsions in each group

[0127]

[0128] Note: - represents not tested

[0129] Experimental Example 4

[0130] The stability of the cinnamaldehyde drug-loaded self-microemulsions prepared in Examples 7, 14 and 15 was investigated.

[0131] The cinnamaldehyde-loaded self-microemulsions prepared in each group were placed in vials and stored in the dark at 4°C. Particle size, PDI, and cinnamaldehyde and drug content were measured by HPLC after 0, 3, 7, 15, 30, 45, and 60 days, respectively. The results are shown in Table 5. As can be seen from the results in the table, the particle size and PDI of the cinnamaldehyde-loaded self-microemulsions did not change significantly within 60 days, and the remaining percentages of cinnamaldehyde and drug did not change significantly.

[0132] Table 5 The results of the storage stability of each group of cinnamaldehyde drug-loaded self-microemulsions

[0133]

[0134] Experimental Example 5: Drug Efficacy Experiment on Rat Liver Fibrosis

[0135] Experimental animals: SD male rats, weighing 180-200 g.

[0136] Construction of an 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 margin was elevated, the duodenum was pulled open, and the common bile duct was separated by 2-3 cm. 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, and the liver was restored to its original position and the incision was sutured. After the animals woke up from anesthesia, they were given a normal diet and free access to water.

[0137] Model group (5 animals): 5 of the above BDL model animals were selected as the BDL model group (abbreviated as BDL). Starting from the second day after surgery, normal saline was administered by gavage once a day.

[0138] Sham-operated group (5 animals): 5 experimental animals were selected as the sham-operated group (abbreviated as Sham). They were fasted for 12 hours before surgery. After anesthesia with isoflurane, the abdomen was opened under sterile conditions, and the incision was sutured. After the animals woke up from anesthesia, they were fed normally and had free access to water. Starting from the second day after surgery, normal saline was administered by gavage once a day.

[0139] Active pharmaceutical ingredient group (10 animals): Ten of the BDL model animals were selected as active pharmaceutical ingredient groups (referred to as CIN and DHI), with 5 animals in each group. Cinnamaldehyde (referred to as CIN) and dihydrotanshinone I (referred to as DHI) were administered daily by gavage starting the day after surgery. Cinnamaldehyde was evenly dispersed in olive oil; dihydrotanshinone I was evenly dispersed in CMC-Na solution. The single dose for both groups was approximately 82 mg / kg / day for cinnamaldehyde and 2.9 mg / kg / day for dihydrotanshinone I.

[0140] Emulsion Group (15 animals): Fifteen of the BDL model animals were selected as the emulsion groups (abbreviated as CR, DR, and CDR). Starting on the second day after surgery, they were gavage-administered once daily with the cinnamaldehyde-loaded self-microemulsion prepared in Example 7 (abbreviated as CDR), the cinnamaldehyde self-microemulsion prepared using the same formulation as in Example 7 (abbreviated as CR), or the dihydrotanshinone I self-microemulsion (abbreviated as DR). The single dose was approximately 82 mg / kg / day for cinnamaldehyde and 2.9 mg / kg / day for dihydrotanshinone I. The dihydrotanshinone I self-microemulsion (abbreviated as DR) was prepared 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%.

[0141] After 14 days of drug administration (or saline), the rats in each group fasted for 12 hours, after which blood and liver tissue samples were collected. Serum was collected for biochemical analysis. The results are shown in Table 6. Compared with the sham group, ALT, AST, ALP, TBA, and TBiLi levels were significantly elevated in the BDL model group. Compared with the BDL model group, the CDR group significantly reduced AST, TBA, and TBiLi levels in rat serum, indicating that cinnamaldehyde-loaded self-microemulsion significantly improved liver function in BDL rats, with superior effects compared to both the API and a single drug-loaded emulsion.

[0142] Table 6 Anti-liver fibrosis efficacy index results

[0143]

[0144] 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.

[0145] 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 in the figure. The results showed that compared with the sham group, the BDL model group showed significant hepatocellular 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 showed no significant improvement in bile duct hyperplasia, hepatocellular necrosis, and collagen deposition in the liver, while CR, DR, and CDR reduced bile duct hyperplasia, hepatocellular necrosis, and collagen deposition. However, some inflammatory infiltration and collagen deposition were still present in the CDR group, indicating that cinnamaldehyde-loaded self-microemulsion has a certain effect on improving the symptoms of liver fibrosis in the middle and late stages, and can delay the progression of liver fibrosis, but cannot reverse it.

[0146] Experimental Example 6: Efficacy of Non-alcoholic Steatohepatitis (NASH)

[0147] Experimental animals: C57BL / 6 male mice, weighing 25-28 g.

[0148] Non-alcoholic steatohepatitis (MCD) animal model: 64 experimental animals were randomly divided into 8 groups, with 8 animals in each group. Each group was given an adaptive diet for the first 3 days. Starting from the fourth day, they were given an MCD diet for 7 weeks to establish NASH model. Continuous drug administration began from the 5th week.

[0149] 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 gavaged with normal saline once a day.

[0150] Control group (8 animals): 8 experimental animals were taken as the control group (abbreviated as Control), and were given a normal diet for 7 consecutive weeks. Starting from the 5th week, they were given normal saline by gavage once a day.

[0151] Active pharmaceutical ingredient group (24 animals): 24 of the MCD model animals were selected as active pharmaceutical ingredient groups, with 8 animals in each group. Starting in week 5, they were gavaged once daily with cinnamaldehyde active pharmaceutical ingredient (CIN), dihydrotanshinone I active pharmaceutical ingredient (DHI), or a physical mixture of cinnamaldehyde and dihydrotanshinone I (MIX). Cinnamaldehyde was evenly dispersed in olive oil; dihydrotanshinone I was evenly dispersed in CMC-Na solution. The single dose for each group was approximately 82 mg / kg / day for cinnamaldehyde and 2.9 mg / kg / day for dihydrotanshinone I.

[0152] Emulsion group (24 animals): 24 of the MCD model animals were selected as the emulsion group. Starting from week 5, they were gavaged once daily with the cinnamaldehyde drug-loaded self-microemulsion (CDR) prepared in Example 7, as well as the cinnamaldehyde nanoemulsion (CR) and dihydrotanshinone I self-microemulsion (DR) prepared in Step 1 of Experimental Example 7 using the same formulation. The single dose for each group was approximately 82 mg / kg / day for cinnamaldehyde and 2.9 mg / kg / day for dihydrotanshinone I.

[0153] After 7 weeks, the mice were fasted for 12 hours, and then blood, liver tissue and other samples were collected. 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, single drug-loaded emulsion and their physical mixture.

[0154] Table 7 Anti-nonalcoholic fatty liver disease efficacy index results 1

[0155]

[0156] Table 8 Anti-nonalcoholic fatty liver disease efficacy index results 2

[0157]

[0158]

[0159] Note: In Table 7 and Table 8, *** indicates significant difference compared with the Control group, P < 0.001; ** indicates significant difference compared with the Control group, P < 0.01; * indicates significant difference compared with the Control group, P < 0.05; ### indicates significant difference compared with the MCD group, P < 0.001; ## indicates significant difference compared with the MCD group, P < 0.01; # indicates significant difference compared with the MCD group, P < 0.05.

[0160] The liver tissue samples of each group were stained with hematoxylin-eosin (HE) and oil red, and the staining results were as follows: Figure 2 As shown. The results showed that compared with the Control group, the MCD model group had a large amount of fat vacuoles accumulation, lipid degeneration and inflammatory infiltration in the liver, 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 the CDR group had no obvious fat vacuoles under the field of view, reversing non-alcoholic fatty liver disease. However, the CIN, DHI and CIN and DHI physical mixture groups had no significant improvement in liver fat accumulation and inflammatory infiltration. In summary, the results show that cinnamaldehyde-loaded self-microemulsion has a significant therapeutic effect on early non-alcoholic fatty liver disease, can promote fat decomposition, reduce the occurrence of inflammation, and reverse non-alcoholic fatty liver disease.

[0161] 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 principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A cinnamaldehyde drug-loaded self-microemulsion, characterized in that: The method is prepared from the following raw materials in parts by weight: 1-4.5 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 drug; When the oil phase is 0 parts, the emulsifier is castor oil polyoxyethylene ether EL or a combination of castor oil polyoxyethylene ether EL + polyoxyethylene hydrogenated castor oil RH, the mass ratio of cinnamaldehyde to emulsifier is 1:0.9-1.1, and the poorly soluble drug is dihydrotanshinone I; When the oil phase is not 0 parts, the emulsifier is castor oil polyoxyethylene ether EL or a combination of castor oil polyoxyethylene ether EL + polyoxyethylene hydrogenated castor oil RH, the mass ratio of cinnamaldehyde to emulsifier is 1:0.9-3.5, the oil phase is medium-chain triglyceride, the mass ratio of oil phase to emulsifier is 1:0.9-3.5, and the insoluble drug is dihydrotanshinone I; When the emulsifier is a combination of polyoxyethylene castor oil ether EL and polyoxyethylene hydrogenated castor oil RH, the mass ratio of polyoxyethylene castor oil ether EL to polyoxyethylene hydrogenated castor oil RH is 1:

1.

2. The method for preparing the cinnamaldehyde drug-loaded self-microemulsion according to claim 1, wherein: The method comprises the following steps: uniformly mixing the weight portions of cinnamaldehyde, oil phase, emulsifier and insoluble drug to obtain cinnamaldehyde drug-loaded self-microemulsion.

3. The preparation method according to claim 2, wherein The mixing speed is 550-650 rpm.

4. Use of the cinnamaldehyde drug-loaded self-microemulsion according to claim 1 or the cinnamaldehyde drug-loaded self-microemulsion obtained by the preparation method according to claim 2 or 3 in preparing a product for improving liver fibrosis.

5. Use of the cinnamaldehyde drug-loaded self-microemulsion according to claim 1 or the cinnamaldehyde drug-loaded self-microemulsion obtained by the preparation method according to claim 2 or 3 in preparing a product for treating non-alcoholic steatohepatitis.

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