Progesterone self-emulsifying formulation, preparation method therefor, and use thereof

Abstract

A progesterone self-emulsifying formulation, a preparation method therefor, and use thereof. The progesterone self-emulsifying formulation comprises the following preparation raw materials by mass fraction: 0.5%-20% of progesterone, 15%-50% of an oily solvent, 45%-80% of a surfactant, and 0-25% of a co-surfactant. Progesterone in the progesterone self-emulsifying formulation is in a completely dissolved state, which promotes the absorption of progesterone in the body, thereby improving the oral bioavailability of progesterone. In addition, by means of dispersing progesterone in a self-emulsifying system, under the action of an oil phase, progesterone enters the lymphatic circulation together with chylomicrons, thereby avoiding the liver first-pass effect, not only increasing the oral bioavailability of progesterone, but also reducing the plasma concentration of metabolites, and reducing the incidence of adverse reactions of oral progesterone.

Description

Progesterone self-emulsifying preparation, preparation method and application thereof

[0001] The present application claims priority to the Chinese patent application No. 202410788143.7, filed on June 18, 2024, and entitled "Progesterone self-emulsifying preparation, preparation method and application thereof", the content of which is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0002] The present application relates to the technical field of pharmaceutical preparations, in particular to a progesterone self-emulsifying preparation, a preparation method and application thereof. BACKGROUND

[0003] Progesterone, also known as progestin, is a kind of endogenous progestogen with the chemical formula of C 21 H 30 O2, and the structural formula as shown in formula 1, is a kind of endogenous progestogen. Progesterone is a very important intermediate product involved in steroid synthesis, and plays a role in the reproductive system, mammary glands, metabolism, etc., and is the main hormone for luteal support and maintenance of pregnancy.

[0004] Progesterone is the main factor affecting the transformation of endometrium from the proliferative phase to the secretory phase, and can promote the maturation, exfoliation of endometrium and maintain the menstrual cycle of women. At the same time, progesterone affects the thickness and blood flow of endometrium, and plays a role in stabilizing endometrium and preventing miscarriage in early pregnancy. As a natural progestogen, progesterone has no adverse effects on the fetus and patients, and is the first choice and main drug for patients to obtain luteal support and maintain pregnancy. It is mainly used in the treatment of threatened abortion caused by luteal dysfunction, menstrual disorders caused by ovulation disorders, perimenstrual syndrome, etc. According to the data released by the China Population Association and the National Health Commission, the rate of infertility of Chinese couples of childbearing age is gradually rising, and there are about 50 million infertile people. With the continuous improvement of the people's demand for reproductive health and the comprehensive opening of the "three-child" policy, the demand for progesterone in maintaining pregnancy and assisted reproduction (ART) is increasing, which greatly promotes the market demand for progesterone. During the luteal support in the ART process, if the patient has luteal dysfunction, it may lead to a decrease in pregnancy rate and an increase in miscarriage rate, thereby affecting the final treatment effect. Therefore, supplementing exogenous progesterone is a crucial link in ART. At present, the progesterone preparations used in assisted reproduction in the clinic mainly include injection, vaginal gel and oral preparations. However, these preparations on the market have some problems, which are difficult to meet the compliance and safety needs of patients in clinical medication.

[0005] Progesterone oil injection is the traditional administration method of progesterone, which has high bioavailability, good absorption and low price. However, due to the fact that oil is not soluble in water and has high viscosity, it is not easy to diffuse and absorb after local injection, and local adverse reactions such as pain, inflammation, induration, swelling and bleeding often occur at the injection site. In addition, due to the long-term use of progesterone injection, the inflammatory and allergic reactions of some patients gradually worsen, and even severe cases require surgical intervention. In addition, progesterone needs to be injected daily, and the daily visits to the hospital by pregnant women lead to poor medication compliance.

[0006] Progesterone vaginal gel plays a role by increasing the local concentration of progesterone in the uterus through vaginal administration, and has high absorption and utilization. However, patients need to maintain a lying position when using it, which is inconvenient, and there are problems such as easy leakage, difficulty in discharging drug residues and poor patient adaptability after administration. The most important product on the market is "Cyclo-F" produced by Merck, which has been in a monopoly position for a long time due to the lack of similar competitors, and the price has been relatively high. The generic products of "Cyclo-F" and new products for vaginal administration need to pass the double evaluation of pharmacokinetic equivalence and pharmacodynamic equivalence, which makes the development of preparations difficult and the cost of clinical trials high.

[0007] Progesterone oral preparations do not require patients to frequently visit hospitals, and the administration method is simple and convenient, which has good patient medication compliance compared with injections and vaginal administration. The progesterone oral products that have been marketed in China include "Angistan (France Besins)", "Yimashin (Zhejiang Xianju Pharmaceutical Co., Ltd.)" and "Qining (Zhejiang Aisheng Pharmaceutical Co., Ltd.)" progesterone capsules. However, the above progesterone oral preparations have significant liver first-pass effect, resulting in oral bioavailability of less than 10% and large individual differences among patients. In addition, the side effects of progesterone after oral administration are affected by the metabolite of progesterone, allopregnanolone. A Allopregnanolone is a neurosteroid and a positive allosteric modulator of GABA A receptors. GABA receptors are the main biological targets of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). When the blood concentration of allopregnanolone reaches a certain concentration, it will cause side effects such as headache, dizziness and fatigue.

[0008] Therefore, there is a great clinical need and market prospect for a progesterone oral preparation with high bioavailability and low toxicity.

[0009] SUMMARY

[0010] Therefore, the purpose of the present application is to provide a progesterone self-emulsifying preparation, a preparation method and application thereof. The progesterone self-emulsifying preparation of the present application has high bioavailability and low toxicity.

[0011] To achieve the above-mentioned application purposes, the present application provides the following technical solutions.

[0012] The present application provides a progesterone self-emulsifying preparation, which comprises the following preparation raw materials in mass fraction:

[0013] Progesterone 0.5% to 20%, oily solvent 15% to 50%, and surfactant 45% to 80%.

[0014] Preferably, the preparation raw materials comprise progesterone 1% to 15%, oily solvent 20% to 50%, and surfactant 50% to 70% in mass fraction.

[0015] Preferably, the preparation raw materials comprise progesterone 1% to 10%, oily solvent 30% to 50%, and surfactant 50% to 70% in mass fraction.

[0016] Preferably, the surfactant is one or more of polyoxyethylene castor oil, hydrogenated polyoxyethylene castor oil, Tween-20, Tween-60, Tween-80, vitamin E-TPGS, and caprylocaproyl macrogol glycerides.

[0017] Preferably, the preparation raw materials of the progesterone self-emulsifying preparation further comprise a co-surfactant, which is one or more of diethylene glycol monoethyl ether, polyglycerol oleate, PEG400, isopropyl alcohol, propylene glycol laurate, ethanol, and 1,2-propanediol; the mass fraction of the co-surfactant is ≤25% based on the mass fraction of the progesterone.

[0018] Preferably, the oily solvent is one or more of soybean oil, rapeseed oil, palm oil, peanut oil, sunflower oil, coconut oil, flaxseed oil, castor oil, perilla oil, corn oil, monolinolein, linolein, trilinolein, monoolein, diolein, triolein, medium-chain triglyceride, caprylocaproyl macrogol glyceride, oleic acid, linoleic acid, ethyl oleate, ethyl linoleate, and palmitic acid.

[0019] The present application also provides a preparation method of the progesterone self-emulsifying preparation described in the above technical solutions, which comprises the following steps:

[0020] Mixing the preparation raw materials to obtain the progesterone self-emulsifying preparation.

[0021] Preferably, the mixing is performed at a temperature of 20 to 100℃ for 0.5 to 4h; and the mixing is performed under stirring.

[0022] The application further provides application of the progesterone self-emulsifying preparation or the progesterone self-emulsifying preparation prepared by the preparation method in the technical solution to progesterone oral preparations.

[0023] Preferably, the dosage form of the progesterone oral preparation is a soft capsule, a liquid capsule, a hard capsule or a dripping pill.

[0024] The application provides a progesterone self-emulsifying preparation, which comprises the following raw materials in mass fraction: 0.5-20% of progesterone, 15-50% of an oily solvent and 45-80% of a surfactant. Beneficial effects:

[0025] (1) The progesterone in the progesterone self-emulsifying preparation provided by the application is in a completely dissolved state, and after oral administration, the progesterone is in micelles, mixed micelles or other carriers in the intestinal tract, which enhances the solubility of the progesterone in the intestinal environment and promotes the absorption of the progesterone in the body, i.e., the bioavailability of the progesterone is improved.

[0026] (2) By dispersing the progesterone in the self-emulsifying system, the progesterone enters the lymphatic circulation together with chylomicrons under the action of the oil phase, thereby avoiding the liver first-pass effect, increasing the oral bioavailability of the progesterone and reducing the generation of metabolites with toxic side effects. Compared with the commercially available French progesterone gel pills (angitrin), the progesterone self-emulsifying preparation has significantly improved oral bioavailability and significantly reduced the concentration of metabolites causing adverse reactions.

[0027] (3) The raw materials required for the preparation of the progesterone self-emulsifying preparation are easy to obtain. BRIEF DESCRIPTION OF DRAWINGS

[0028] Fig. 1 is a particle size distribution diagram of the progesterone self-emulsifying preparation prepared in Example 4;

[0029] Fig. 2 is a ternary phase diagram of caprylic / capric triglyceride, Tween-80 and Transcutol HP;

[0030] Fig. 3 is a drug-time curve diagram of rats orally administered with the progesterone self-emulsifying preparation obtained in Example 2 and angitrin;

[0031] Fig. 4 is a blood drug concentration change diagram of mice administered with angitrin, the progesterone self-emulsifying preparation obtained in Example 5 and the progesterone self-emulsifying preparation obtained in Example 5 and milbemycin;

[0032] Fig. 5 is a progesterone drug-time curve of beagles orally administered with the progesterone self-emulsifying preparation obtained in Example 2 and angitrin;

[0033] Figure 6 is a plot of the concentration-time curve of metabolite allopregnanolone after oral administration of progesterone self-emulsifying formulation of Example 2 and androstanediol to beagle dogs. DETAILED DESCRIPTION

[0034] The present application provides a progesterone self-emulsifying formulation, which comprises the following raw materials by mass fraction:

[0035] Progesterone 0.5% to 20%, oily solvent 15% to 50%, surfactant 45% to 80%.

[0036] In the present application, the raw materials used in the present application are preferably commercially available products, unless otherwise specified.

[0037] The progesterone self-emulsifying formulation provided by the present application comprises progesterone by mass fraction of 0.5% to 20%, preferably 1% to 15%, further preferably 1% to 10%, most preferably 2% to 5%, and specifically preferably 2.0%, 2.5%, 3.0% or 4.0%.

[0038] The progesterone self-emulsifying formulation provided by the present application comprises oily solvent by mass fraction of 15% to 50% based on the mass fraction of progesterone, preferably 20% to 50%, further preferably 30% to 50%, and specifically preferably 19.2%, 19.5%, 28.8%, 29.1%, 29.3%, 29.4%, 38.4% or 38.8%. In the present application, the oily solvent is preferably one or more of soybean oil, rapeseed oil, palm oil, peanut oil, sunflower oil, coconut oil, flaxseed oil, castor oil, perilla oil, corn oil, monolinolein, linolein, trilinolein, monoolein, diolein, triolein, medium-chain triglyceride, caprylocapryl triglyceride, oleic acid, linoleic acid, ethyl oleate, ethyl linoleate and palmitic acid, and further preferably ethyl linoleate, ethyl oleate-monolinolein, monolinolein, caprylocapryl triglyceride, caprylocapryl triglyceride-monolinolein, monoolein, ethyl oleate, linolein, linolein-trilinolein, diolein-triolein. The selected oily solvent of the present application can improve the dispersibility of the progesterone drug in the gastrointestinal tract, thereby improving the oral bioavailability of the progesterone drug. In addition, the oily solvent of the present application can promote the lymphatic absorption of progesterone, thereby reducing its liver first-pass effect, reducing the concentration of metabolite allopregnanolone in the body, and further reducing the side effects of the progesterone self-emulsifying formulation.

[0039] The preparation raw material of the progesterone self-emulsifying preparation provided in the present application comprises a surfactant with a mass fraction of 45% to 80% of progesterone, preferably 50% to 70%, and specifically preferably 48.2%, 48.5%, 52.8%, 53.7%, 57.60%, 58.2%, 58.8%, 67.2% or 67.3%. In the present application, the surfactant is preferably one or more of polyoxyethylene castor oil, hydrogenated polyoxyethylene castor oil, Tween-20, Tween-60, Tween-80, vitamin E-TPGS and caprylocaproyl macrogol glycerides, and is further preferably Tween-20, polyoxyethylene castor oil-caprylocaproyl macrogol glycerides, caprylocaproyl macrogol glycerides, Tween-80, hydrogenated polyoxyethylene castor oil, Tween-60. The selection of the surfactant can reduce the surface tension and form a monolayer between the oil phase and the aqueous phase, so that the progesterone stays in the gastrointestinal tract (GIT) in a soluble form, which is beneficial to the absorption of the drug by the intestinal epithelium.

[0040] The preparation raw material of the progesterone self-emulsifying preparation provided in the present application further comprises a co-surfactant, which is preferably one or more of diethylene glycol monoethyl ether, polyglycerol oleate, PEG400, isopropyl alcohol, propylene glycol laurate, ethanol and 1,2-propanediol, and is more preferably diethylene glycol monoethyl ether, polyglycerol oleate, propylene glycol laurate, 1,2-propanediol, PEG400. In the present application, the mass fraction of the co-surfactant is preferably ≤25%, further preferably 0.1% to 25%, more preferably 0.1% to 15%, most preferably 0.1% to 5%, and specifically preferably 4.8%, 9.5%, 9.6%, 9.7%, 9.8%, 20.0% or 24.3%, based on the mass fraction of the progesterone. In the present application, the co-surfactant can increase the drug loading capacity of the progesterone self-emulsifying preparation and enhance the dispersion capacity of the drug in the progesterone self-emulsifying preparation by increasing the interfacial fluidity.

[0041] The present application also provides a preparation method of the progesterone self-emulsifying preparation described in the above technical solutions, comprising the following steps:

[0042] Mixing the preparation raw materials to obtain the progesterone self-emulsifying preparation.

[0043] In the present application, the temperature of the mixing is preferably 20 to 100°C, and the time is preferably 0.5 to 4h; the mixing is performed under stirring, and the stirring mode is preferably magnetic stirring.

[0044] The preparation method provided in the present application is very simple, only needs to mix the preparation raw materials, is convenient for large-scale process production, and the prepared progesterone self-emulsifying preparation is relatively stable. The progesterone self-emulsifying preparation prepared in the present application can be directly used for the production of soft capsules and dripping pills, which is beneficial to the industrialized production.

[0045] The application also provides the use of the progesterone self-emulsifying preparation described in the technical solution or the progesterone self-emulsifying preparation prepared by the preparation method in the technical solution in the preparation of a progesterone oral preparation.

[0046] In the application, the dosage form of the progesterone oral preparation is preferably a soft capsule, a liquid capsule, a hard capsule or a dripping pill.

[0047] The progesterone self-emulsifying preparation, the preparation method and the application thereof provided in the application are described in detail below in combination with examples, but they should not be understood as limitations to the protection scope of the application.

[0048] Example 1

[0049] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0050] Progesterone 4.0 wt%, ethyl linoleate 19.2 wt%, Tween-20 67.2 wt%, diethylene glycol monoethyl ether 9.6 wt%.

[0051] The preparation process is as follows: 40.0 mg of progesterone, 192 mg of ethyl linoleate, 672 mg of Tween-20 and 96 mg of diethylene glycol monoethyl ether are weighed in a round-bottom flask, and the progesterone self-emulsifying preparation is obtained after magnetic stirring at 40°C for 30 min.

[0052] Example 2

[0053] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0054] Progesterone 3.0 wt%, ethyl oleate 20.4 wt%, glyceryl monolinoleate 18.4 wt%, polyoxyethylene castor oil 20.4 wt%, caprylocaproyl macrogol glycerides 28.1 wt%, polyglyceryl oleate 9.7 wt%.

[0055] The preparation process is as follows: 30.0 mg of progesterone, 204.0 mg of ethyl oleate, 184.0 mg of glyceryl monolinoleate, 204 mg of polyoxyethylene castor oil, 281 mg of caprylocaproyl macrogol glycerides and 97.0 mg of polyglyceryl oleate are weighed in a round-bottom flask, and the progesterone self-emulsifying preparation is obtained after magnetic stirring at 40°C for 30 min.

[0056] Example 3

[0057] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0058] Progesterone 4.0 wt%, glyceryl monolinoleate 38.4 wt%, caprylocaproyl macrogol glycerides 52.8 wt%, polyglyceryl oleate 4.8 wt%.

[0059] The preparation process is: taking 40.0 mg of progesterone, 384.0 mg of monolinoleate glyceride, 528.0 mg of caprylic capric acid polyethylene glycol glyceride and 48.0 mg of polyglyceryl oleate in a round-bottom flask, stirring at 40°C for 30 min to obtain a progesterone self-emulsifying preparation.

[0060] Example 4

[0061] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0062] Progesterone 4.0 wt%, caprylic capric acid triglyceride 19.2 wt%, Tween-80 67.3 wt%, diethylene glycol monoethyl ether 9.5 wt%.

[0063] The preparation process is: taking 40.0 mg of progesterone, 192.0 mg of caprylic capric acid triglyceride, 673.0 mg of Tween-80 and 95 mg of diethylene glycol monoethyl ether in a round-bottom flask, stirring at 40°C for 30 min to obtain a progesterone self-emulsifying preparation.

[0064] The obtained progesterone self-emulsifying preparation is diluted with water to 100 times, and the particle size is measured by a particle size instrument. The results are shown in Figure 1. As can be seen from Figure 1, the particle size is 149.10±1.17 nm, and the PDI is 0.275±0.007.

[0065] Example 5

[0066] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0067] Progesterone 2.5 wt%, caprylic capric acid triglyceride 9.7 wt%, monolinoleate glyceride 9.8 wt%, Tween-80 53.7 wt%, diethylene glycol monoethyl ether 24.3 wt%.

[0068] The preparation process is: taking 38.8 g of caprylic capric acid triglyceride, 39.2 g of monolinoleate glyceride, 214.8 g of Tween-80 and 97.2 g of diethylene glycol monoethyl ether according to the selected prescription ratio, stirring uniformly by magnetic stirring, then adding 10.0 g of progesterone raw material, continuing to stir by magnetic stirring at 45°C until the solution is clear, to obtain a progesterone self-emulsifying preparation.

[0069] Example 6

[0070] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0071] Progesterone 2.5 wt%, monolinoleate glyceride 29.3 wt%, hydrogenated polyoxyethylene castor oil 48.2 wt%, lauric acid propylene glycol ester 20.00 wt%.

[0072] The preparation process is as follows: taking 25.0 mg of progesterone, 293.0 mg of glyceryl monooleate, 482.0 mg of hydrogenated polyoxyethylene castor oil, and 200.0 mg of propylene glycol laurate in a round-bottom flask, and stirring magnetically at 40°C for 30 min to obtain a progesterone self-emulsifying preparation.

[0073] Example 7

[0074] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0075] Progesterone 2.0 wt%, ethyl oleate 29.4 wt%, Tween-80 58.8 wt%, 1,2-propylene glycol 9.8 wt%.

[0076] The preparation process is as follows: taking 20.0 mg of progesterone, 294.0 mg of ethyl oleate, 588.0 mg of Tween-80, and 98.0 mg of 1,2-propylene glycol in a round-bottom flask, and stirring magnetically at 40°C for 30 min to obtain a progesterone self-emulsifying preparation.

[0077] Example 8

[0078] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0079] Progesterone 3.0 wt%, glyceryl dilinoleate 29.1 wt%, Tween-60 58.2 wt%, PEG400 9.7 wt%.

[0080] The preparation process is as follows: taking 30.0 mg of progesterone, 291.0 mg of glyceryl dilinoleate, 582.0 mg of Tween-60, and 97.0 mg of PEG400 in a round-bottom flask, and stirring magnetically at 40°C for 30 min to obtain a progesterone self-emulsifying preparation.

[0081] Example 9

[0082] The raw materials for preparing the progesterone self-emulsifying preparation include:

[0083] Progesterone 4.00 wt%, glyceryl dilinoleate 14.40 wt%, glyceryl trilinoleate 14.40 wt%, Tween-60 57.60 wt%, 1,2-propylene glycol 9.60 wt%.

[0084] The preparation process is as follows: taking 40.0 mg of progesterone, 144.0 mg of glyceryl dilinoleate, 144.0 mg of glyceryl trilinoleate, 576.0 mg of Tween-60, and 96.0 mg of 1,2-propylene glycol in a round-bottom flask, and stirring magnetically at 40°C for 30 min to obtain a progesterone self-emulsifying preparation.

[0085] Example 10

[0086] The preparation raw materials of the progesterone self-emulsifying formulation include:

[0087] Progesterone 4.00 wt%, glyceryl dioleate 14.40 wt%, glyceryl trioleate 14.40 wt%, Tween-60 57.60 wt%, 1,2-propanediol 9.60 wt%.

[0088] The preparation process is as follows: 40.0 mg of progesterone, 144.0 mg of glyceryl dioleate, 144.0 mg of glyceryl trioleate, 576.0 mg of Tween-60, and 96.0 mg of 1,2-propanediol are weighed in a round-bottom flask, and the progesterone self-emulsifying formulation is obtained after magnetic stirring at 40°C for 30 min.

[0089] Test Example 1: Ternary phase diagram of caprylic / capric triglyceride, Tween-80, and diethylene glycol monoethyl ether three kinds of excipients

[0090] The ratio of Example 3 is screened by making a ternary phase diagram of caprylic / capric triglyceride, Tween-80, and diethylene glycol monoethyl ether three kinds of excipients, and the excipient ratio that is nanoemulsion after emulsification is screened.

[0091] The mass concentration of the fixed co-surfactant in the system is 0%, 5%, 10%, 15%, 20%, 25%, and 30%, and the corresponding mass concentration of the surfactant is 40%, 45%, 50%, 55%, 60%, 65%, and 70% at each concentration, and the oil phase concentration is added to 100%. Different excipients are weighed according to the above concentration ratio, vortexed uniformly, and then the blank emulsion is added to 100 times the volume of water, and constant speed stirring is carried out on a magnetic stirrer for 2 min. The self-emulsifying zone is determined according to the clarity of the emulsion. Pseudo-ternary phase diagrams are made with the oil phase, surfactant, and co-surfactant as the apex, and the experimental results are shown in FIG. 2. FIG. 2 is a ternary phase diagram of caprylic / capric triglyceride, Tween-80, and diethylene glycol monoethyl ether (Transcutol HP) three kinds of excipients. As can be seen from FIG. 2, the oil phase, surfactant, and co-surfactant need to be in a certain ratio to cause self-emulsification.

[0092] Test Example 2: Changes in progesterone and allopregnanolone concentrations in rat plasma after intragastrically administering the progesterone self-emulsifying formulation prepared in Example 2 and Angiotensin

[0093] The progesterone self-emulsifying formulation prepared in Example 2 is subjected to pharmacokinetic analysis, and the oral bioavailability thereof is calculated.

[0094] Select 24 female SD rats, randomly divided into A, B two groups, 12 in each group, before the experiment fasting without water. A group of single dose of 180 mg / kg progesterone gavage given progesterone soft capsules (Angelica), B group single dose given progesterone dose of 156 mg / kg progesterone self-emulsifying preparation in the application of example 2, before and after administration at different time points, the orbital blood was placed in heparin sodium tube, 3500 rpm centrifugation 10 min, separation of plasma.

[0095] Plasma sample analysis: rat plasma sample processing takes the ammonium sulfate-acetonitrile two-phase extraction method, the operation steps are as follows: take 50 μL of plasma sample, add 10 μL of internal standard working solution (1 μg / mL progesterone-D9 working solution) and 25 μL of saturated ammonium sulfate solution, vortex mix, then add 200 μL of acetonitrile, vortex for about 1 min, centrifuge for 10 min (4℃, 20,200 g), take 100 μL of supernatant, add 100 μL of water, vortex (about 30 s), take 100 μL of sample for LC-MS / MS analysis.

[0096] Figure 3 is a rat oral progesterone self-emulsifying preparation of example 2 and Angelica time curve diagram, from figure 3 can be seen: compared with Angelica, rat oral progesterone self-emulsifying preparation of example 2 in vivo plasma progesterone concentration increased significantly.

[0097] Table 1 rat gavage Angelica and example 2 after the main PK parameters of progesterone in plasma

[0098] Note: the above data is the statistical moment parameter obtained by DAS2.0 analysis.C max : peak concentration; T max : peak time; t 1 / 2 :

[0099] half-life; AUC: area under the curve. AUC dose normalized adjustment.

[0100] ** p<0.01, compared with Angelica group.

[0101] Table 1 is the rat gavage Angelica and example 2 after the main PK parameters of progesterone in plasma. Compared with the reference preparation Angelica, oral progesterone self-emulsifying preparation of example 2 t 1 / 2 There is no significant change, which shows that the progesterone self-emulsifying preparation of example 2 has no significant effect on the metabolism of progesterone in vivo. Oral progesterone self-emulsifying preparation of example 2 and Angelica C max There is a significant gap, progesterone self-emulsifying preparation of example 2 will increase the C maxfrom 36.35 ± 18.54 ng / mL to 247.84 ± 317.75 ng / mL. Since the dosages of anastrozole and progesterone self-emulsifying formulation obtained in Example 2 were different, the AUCs need to be normalized by dosage, and calculated based on the dosage of anastrozole. The progesterone AUC of the anastrozole group was 99.85 ± 42.83 ng / mL·h, and the progesterone AUC of the Example 2 group was 293.43 ± 134.05 ng / mL·h, and the progesterone AUCs of the two groups had a significant difference (p < 0.01). The relative oral bioavailability of progesterone of Example 2 was 293.87% of anastrozole. It can be seen that the blood drug concentration of progesterone in the body after oral administration of anastrozole is low, and the bioavailability is poor. In comparison, Example 2 can significantly improve the blood drug concentration of progesterone in the body and improve the oral absorption of progesterone. 0-t 0-t 0-t

[0102] Investigation of the absorption mechanism of the progesterone self-emulsifying formulation of Test Example 3

[0103] To investigate the overall absorption-promoting effect of progesterone self-emulsifying formulation on progesterone in animals, the pharmacokinetics in vivo was investigated by gavage administration of ICR mice. At the same time, a mouse chylomicron blocking model was established to study the influence of lymphatic transport on the improvement of progesterone absorption and oral bioavailability.

[0104] Experimental scheme: Before the experiment, the ICR mice were fasted with free water. 36 mice were randomly divided into A1, A2, B1, B2, C1, C2 groups, 6 mice in each group. A1, A2 groups were given anastrozole by gavage, and blood was taken at intervals; B1, B2 groups were given the progesterone self-emulsifying formulation obtained in Example 5 by gavage, and blood was taken at intervals; C1, C2 groups were given anastrozole by gavage after intraperitoneal injection of cycloheximide 1 hour before administration to establish a chylomicron blocking model, and blood was taken at intervals. The progesterone gavage dose of the 6 groups of mice was 200 mg / kg.

[0105] According to the grouping, the mice of A1 group, B1 group and C1 group were taken blood from the orbit at 0.5, 1.5, 4 and 8 hours after gavage administration, respectively, and the mice of A2 group, B2 group and C2 group were taken blood from the orbit at 1, 2, 6 and 12 hours after gavage administration, respectively. The obtained blood samples were placed in heparin tubes and immediately centrifuged at 400g for 10 min, and the supernatant was taken to obtain plasma samples and stored at -20°C.

[0106] ​​​Plasma sample analysis: 100 μL of plasma sample was taken, 100 μL of saturated ammonium sulfate solution was added and vortexed for 1 min, 400 μL of methanol was added, vortexed for 1 min and centrifuged at 20,200 g for 10 min. The supernatant was taken and placed in a new 1.5 mL centrifuge tube, dried by nitrogen blowing at 50°C, 100 μL of methanol was added, vortexed for 3 min, centrifuged at 20,200 g for 10 min, and the supernatant was taken for HPLC analysis. The results are shown in Figure 4, which is a graph showing the changes in blood drug concentration of mice administered with anecortave, the progesterone self-emulsifying preparation of Example 5 and the progesterone self-emulsifying preparation of Example 5 and rapamycin. As shown in Figure 4, the bioavailability of Example 5 was significantly higher than that of anecortave after a single dose (200 mg / kg) of oral administration of Example 5 and anecortave (reference preparation) in mice.

[0107] Table 2. Main PK parameters of progesterone in plasma of mice with chylomicron blocking model and normal mice after oral administration of anecortave and Example 5

[0108] Note: The above data are statistical moment parameters obtained by DAS 2.0 analysis. max : peak concentration; T max : time to peak; t 1 / 2 : half-life; AUC: area under the curve; MRT: mean residence time; CL z : clearance rate.

[0109] * p < 0.05, ** p < 0.01, compared with the anecortave group;

[0110] ▲ p < 0.05, compared with the Example 5 group.

[0111] The pharmacokinetic parameters are shown in Table 2: the progesterone C max of the Example 5 group was 8.58 μg / mL, and the progesterone C max of the anecortave group was 8.14 μg / mL. Although the progesterone C max of Example 5 was not improved, the progesterone AUC of the Example 5 group was 382.52% of that of the anecortave group, i.e. the relative oral bioavailability of progesterone was increased by 3.82 times compared with anecortave. It can be seen that the progesterone self-emulsifying preparation of Example 5 can significantly promote the oral absorption of progesterone. In the chylomicron blocking model, the bioavailability of Example 5 was lower than that of normal mice after a single dose (200 mg / kg) of oral administration in mice, and the relative bioavailability of progesterone was only 35.76% of that of normal mice, i.e. the generation of chylomicrons can inhibit the oral absorption of progesterone. The experimental results can preliminarily prove that Example 5 mainly promotes lymphatic absorption to improve the oral bioavailability of progesterone.

[0112] Test Example 4 Changes in progesterone and allopregnanolone concentrations in the plasma of beagle dogs after oral administration of Example 2 and angusta

[0113] The bioavailability of Example 2 was proved to be greatly improved compared with angusta through the pharmacokinetic results of rats. On this basis, in order to verify the absorption-promoting effect of Example 2, further pharmacokinetic study of Example 2 was carried out on beagle dogs.

[0114] A total of 5 male beagle dogs, weighing 9.66±0.78 kg, were divided into two groups for cross-over experiment, with a washout period of 1 week. The beagle dogs were fasted but not water-deprived for 12 h before the experiment. One group was fed with 2 enteric-coated capsules of Example 2 (94.65 mg of progesterone), and one group was fed with 1 angusta (104.65 mg of progesterone). About 1 mL of blood was collected from the front limb vein before administration and at 5, 10, 20, 30, 45 min and 1, 1.5, 2, 3, 4, 6, 8 h after administration into an anticoagulant tube, centrifuged at 2,000 g (4°C) for 10 min, and the supernatant was stored in a -70°C refrigerator for subsequent LC-MS / MS analysis.

[0115] Plasma sample processing: The beagle dog plasma samples were processed by organic solvent protein precipitation method, and the operation steps were as follows: 200 μL of plasma sample was taken, 40 μL of internal standard working solution (500 ng / mL) was added, vortexed and mixed, then 600 μL of acetonitrile was added, vortexed for about 1 min, centrifuged at 4°C for 10 min at 20,200 g, 700 μL of supernatant was taken and dried, re-dissolved with 100 μL of water containing 50% acetonitrile, vortexed (about 30 s), centrifuged at 4°C for 10 min at 20,200 g, and 100 μL of sample was taken for LC-MS / MS analysis.

[0116] Table 3 Main PK parameters of progesterone in the plasma of beagle dogs after oral administration of angusta and Example 2

[0117] Note: The above data are statistical moment parameters obtained by DAS2.0 analysis. max : peak concentration; T max : time to peak; t 1 / 2 :

[0118] half-life; AUC: area under the curve. AUC was adjusted by dose normalization.

[0119] ** p<0.01, compared with the angusta group.

[0120] Figure 5 shows the pharmacokinetic (PK) curves of progesterone and acetaminophen after oral administration of the self-emulsified progesterone formulation obtained in Example 2 to beagle dogs. Table 3 shows the main PK parameters of progesterone in plasma after gavage administration of acetaminophen and the self-emulsified progesterone formulation obtained in Example 2. After dose normalization, the AUC of progesterone in Example 2 was calculated. 0-t The AUC of progesterone in Angelica was 100.20 ± 46.41 ng / mL·h. 0-t The bioavailability was 12.56 ± 16.59 ng / mL·h, which means that the bioavailability of progesterone in Example 2 was 797.77% of that of Angelica.

[0121] Table 4. Main pharmacokinetic parameters of allogestranolone in plasma of rats after gavage administration of angelon and in Example 2.

[0122] Note: The above data are statistical moment parameters obtained from DAS2.0 analysis. max Peak concentration; T max Peak time;

[0123] t 1 / 2 Half-life; AUC: Area under the curve. AUC is adjusted for dose normalization.

[0124] **p<0.01, compared with the Anqitan group.

[0125] Figure 6 shows the concentration-time curves of allogeneic ketone, a metabolite of progesterone obtained in Example 2, after oral administration of the self-emulsified progesterone preparation and angelica in beagle dogs. Table 4 shows the main pharmacokinetic parameters of allogeneic ketone in plasma of rats after gavage administration of angelica and the self-emulsified progesterone preparation obtained in Example 2. After dose normalization, the AUC of allogeneic ketone in Example 2 was calculated. 0-t The AUC of allogeneic alcohol ketone in Angelica was 25.62 ± 6.07 ng / mL·h. 0-t The concentration was 14.97 ± 3.11 ng / mL·h. Calculate the AUC of allogeneic ketone. 0-t AUC of progesterone 0-t The ratio of allogeneic alcohol ketone to progesterone AUC was found to be only 21.85% of that of oral progesterone ketone after oral administration of the progesterone self-emulsifying preparation obtained in Example 2. This indicates that to achieve the same progesterone AUC, the allogeneic alcohol ketone produced by oral administration of the progesterone self-emulsifying preparation obtained in Example 2 is only 21.85% of that produced by oral administration of progesterone ketone ketone. This result may be because Example 2 allows a large amount of progesterone to enter the lymphatic circulation, avoiding the first-pass effect in the liver and reducing the generation of metabolites. This suggests that the progesterone self-emulsifying preparation obtained in Example 2 has the ability to reduce the incidence of adverse reactions after oral administration of progesterone.

[0126] The above merely describes the preferred embodiments of the present application, and it should be pointed out that, for those skilled in the art, some improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

[0127] The above merely describes the preferred embodiments of the present application, and it should be pointed out that, for those skilled in the art, some improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A self-emulsifying formulation of progesterone, characterized by, The preparation raw materials comprise the following mass fractions: Progesterone 0.5%-20%, oily solvent 15%-50%, and surfactant 45%-80%.

2. The progesterone self-emulsifying formulation according to claim 1, characterized in that, The preparation raw materials comprise the following mass fractions: progesterone 1%-15%, oily solvent 20%-50%, and surfactant 50%-70%.

3. The progesterone self-emulsifying formulation according to claim 2, characterized in that, The preparation raw materials comprise the following mass fractions: progesterone 1%-10%, oily solvent 30%-50%, and surfactant 50%-70%.

4. The progesterone self-emulsifying formulation according to any one of claims 1 to 3, characterized in that, The surfactant is one or more of polyoxyethylene castor oil, hydrogenated polyoxyethylene castor oil, Tween-20, Tween-60, Tween-80, vitamin E-TPGS, and caprylocaproyl macrogol glycerides.

5. The progesterone self-emulsifying formulation according to any one of claims 1 to 3, characterized in that, The preparation raw materials of the progesterone self-emulsifying preparation further comprise a co-surfactant, which is one or more of diethylene glycol monoethyl ether, polyglycerol oleate, PEG400, isopropyl alcohol, propylene glycol laurate, ethanol, and 1,2-propanediol; the mass fraction of the co-surfactant is ≤25%.

6. The progesterone self-emulsifying formulation according to claim 5, characterized in that, The co-surfactant has a mass fraction of 0.1-25%.

7. The progesterone self-emulsifying formulation according to any one of claims 1 to 3, characterized in that, The oily solvent is one or more of soybean oil, rapeseed oil, palm oil, peanut oil, sunflower oil, coconut oil, linseed oil, castor oil, perilla oil, corn oil, monolinolein, dilinolein, trilinolein, monoolein, diolein, triolein, medium-chain triglyceride, caprylocapryl triglyceride, oleic acid, linoleic acid, ethyl oleate, ethyl linoleate, and palmitic acid.

8. A process for the preparation of a self-emulsifying formulation of progesterone according to any one of claims 1 to 7, characterized in that, The method comprises the following steps: The preparation raw materials are mixed to obtain the progesterone self-emulsifying preparation.

9. The production method according to claim 8, characterized by, The mixing is performed at a temperature of 20-100°C for 0.5-4h under stirring.

10. Use of the progesterone self-emulsifying preparation of any one of claims 1-7 or the progesterone self-emulsifying preparation obtained by the preparation method of any one of claims 8-9 in the preparation of a progesterone oral preparation.

11. Use according to claim 10, characterized in that, The dosage form of the progesterone oral preparation is a soft capsule, a liquid capsule, a hard capsule, or a dripping pill.

12. An oral preparation of progesterone, characterized by, The progesterone oral preparation comprises the progesterone self-emulsifying preparation of any one of claims 1-7 or the progesterone self-emulsifying preparation obtained by the preparation method of any one of claims 8-9.

13. The progesterone oral formulation according to claim 12, wherein The dosage form of the progesterone oral preparation is a soft capsule, a liquid capsule, a hard capsule, or a dripping pill.

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