Progesterone-containing pharmaceutical composition and use

Abstract

Disclosed are a progesterone-containing pharmaceutical composition and use, which belong to the technical field of pharmaceutical preparations. The pharmaceutical composition comprises progesterone as an active ingredient, a sustained-release carrier, a pharmaceutically acceptable solvent, and optionally a lipid-soluble non-ionic surfactant. The pharmaceutical composition possesses long-acting sustained-release properties, enables enhanced therapeutic efficacy, significantly reduces dosing frequency and injection-induced irritation, is projected to significantly improve patient compliance in clinical applications, and effectively reduces therapeutic operations.

Description

Pharmaceutical composition containing progesterone and its application Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and more specifically, relates to a pharmaceutical composition containing progesterone and its application. Background Art

[0002] Progesterone is a natural progestogen secreted by the corpus luteum of the ovary and the placenta. It is a white or off-white crystalline powder, odorless, highly soluble in chloroform, soluble in ethanol, ether, or vegetable oil, and insoluble in water. Progesterone is widely used in all stages of pregnancy, including luteal phase support in assisted reproductive technology, threatened abortion, recurrent miscarriage, and prevention of premature birth. As gestational age increases, the demand for assisted reproduction increases. As one of the main progestogens used in assisted reproduction, the clinical demand for progesterone is also increasing.

[0003] The incidence of infertility in my country has continued to rise since 1995. In 1995, the infertility rate was only about 3%. Between 2012 and 2019, the number of infertile people in my country rapidly increased from 40 million to 50 million, and the overall infertility rate rose to 16.4%. It can be clearly seen that the rate of increase in infertility in my country has accelerated in recent years. In 2020, the infertility rate in my country was approximately 16.9%, and Sullivan's data showed that by 2023, the infertility rate of couples of childbearing age in my country will reach 18.2%. With the opening of my country's fertility policy, the desire to have children has been further stimulated. Progesterone is a BCS Class IV drug (low solubility, low permeability). Currently, the progesterone preparations available on the market mainly include oral capsules, oil-soluble injections, and vaginal gels. Oral progesterone is limited by low solubility and first-pass effect in the liver, and its bioavailability is only about 5% (Stanczyk FZ, Hapgood JP, Winer S, et al. Progestogens used in postmenopausal hormone therapy: differences in their pharmacological properties, intracellular actions, and clinical effects [J]. Endocr Rev, 2013, 34 (2): 171-208). After oral administration, the plasma concentration of progesterone is significantly lower than that of intramuscular injection and is very unstable. The blood concentration reaches a peak 1-3 hours after oral administration, then gradually decreases, and the blood concentration is unstable, disappearing completely in about 72 hours. Due to its low bioavailability, a large dose is often required, resulting in significant side effects, including dizziness, drowsiness, and liver damage. Current studies have shown that oral progesterone capsules cannot fully support endometrial development, and the effectiveness of luteal support in assisted reproduction is lower than that of intramuscular and vaginal progesterone injections, while the side effects are increased compared to intramuscular and vaginal administrations. Therefore, oral progesterone capsules are not recommended as a routine luteal support drug in IVF. Progesterone injection uses oil as a solvent. After intramuscular injection, the blood concentration of the drug increases significantly, reaches a peak value in 6-8 hours, and then gradually decreases, which can last for 24 hours and disappear in 48 hours. The injection site is highly irritating and prone to forming local nodules. Occasionally, local abscesses and damage to the sciatic nerve may occur. The absorption and recovery of local nodules and sterile abscesses takes a long time and are difficult to eliminate. At the same time, due to the solvent diffusion effect, the drug is directly released, which not only often causes local pain and irritation at the injection site, but also causes pain and irritation at the injection site due to the peak concentration (C max) or too high blood concentration, causing adverse reactions such as headaches and abdominal pain. When progesterone is used for assisted reproduction, according to the guidelines, it needs to be injected 40-60mg per day, and the injection should be continued for more than 8 weeks. Long-term and high-frequency injections cause great inconvenience to the patient's life. Progesterone vaginal gel The launch of provides patients with an additional medication option, in which the progesterone content is 8%. After the product is administered via the vaginal route, it is rapidly absorbed by vaginal epithelial cells and diffuses to the cervix and uterine body, and completes the diffusion from the endometrium to the myometrium. Although it improves the therapeutic effect to a certain extent, it is easy to cause adverse reactions such as vaginal bleeding and abdominal pain. A randomized controlled study led by Peking University Third Hospital showed that the use of The incidence of luteal phase bleeding in the progesterone injection group was approximately 12.14%, much higher than that in the progesterone injection group (approximately 3%). When bleeding occurs during assisted reproduction, clinically, vaginal administration must be discontinued and replaced with injections. This not only poses corresponding safety risks to the patient, but also places great psychological pressure on both the patient and the medical staff. Although it greatly improves the convenience of drug administration, its bioavailability is only about 5%, so in order to ensure clinical effect, it needs to be taken twice a day. The main ingredients in the formula that play a sustained-release role are polycarbophil and carbomer, which will gradually melt and be discharged from the body in the vaginal environment, causing inconvenience to the patient. Considering the above physiological structure and long-term medication, although While this approach addresses the compliance issues associated with frequent progesterone injections (irritation and daily, continuous dosing), it still creates significant inconvenience for work and daily life, and it doesn't address the need for a reduced dosing frequency, making it an unsuitable option for long-term medication. Therefore, reducing dosing frequency while also minimizing injection irritation, thereby improving patient compliance, is a key clinical issue that urgently needs to be addressed.

[0004] In order to reduce the irritation of drug administration, the patent document of Chinese patent application publication number CN102397255A discloses a progesterone ethosome and its preparation method, wherein progesterone, lipid material, cholesterol and short-chain alcohol are mixed to prepare an alcohol phase, a nonionic surfactant (Tween-60, Tween-80, benzyl alcohol-35, sodium cholate, etc.) and water are mixed to prepare an aqueous phase, the alcohol phase is added to the aqueous phase to obtain colostrum, and then high-pressure emulsification, secondary emulsification, cooling and solidification are performed to obtain progesterone ethosome, and the proportion of water in the system is about 50%. The patent shows that the prepared progesterone ethosome has a high encapsulation rate, a small particle size, fast transdermal absorption, and a high permeation rate, which is beneficial to improving the bioavailability and therapeutic index of the drug, while avoiding the use of organic solvents that are harmful to the human body and having low irritation. However, the ethosome has the following shortcomings: (1) The progesterone content is only 0.05-0.5%, and the drug loading is low, which means that the administration volume is relatively large, which is not convenient for clinical use. (2) In the in vitro cumulative release experiment of the ethosome, a significant burst release phenomenon occurred at 1 hour, and the cumulative drug release rate reached 90% after only 12 hours. Therefore, this existing technology is difficult to solve the problem of reducing the frequency of progesterone administration.

[0005] To improve patient compliance, Chinese patent application publication number CN109223722A discloses a method for preparing a progesterone nanocrystal injection. The progesterone raw material is emulsified to obtain an oil-in-water emulsion, which is then directly freeze-dried and reconstituted to form nanocrystals. Nanocrystal injections often have physical stability issues, including sedimentation, agglomeration, crystal growth, and crystal changes, which may lead to changes in drug release behavior. Although nanocrystals are one of the optional technologies for long-acting preparations and may reduce the frequency of progesterone administration, nanocrystals also require sterile production, expensive production equipment and facilities, complex processes, and relatively small batch sizes, making it difficult to meet current market demand.

[0006] In order to achieve the slow release of progesterone, the Chinese patent application publication number CN113520990A discloses a progesterone thermosensitive gel injection and its preparation method, wherein progesterone is included in the cavity of cyclodextrin to form a complex and dissolved in water, and then the complex is mixed with a thermosensitive gel (i.e., poloxamer or chitosan / carboxymethyl chitosan / sodium glycerophosphate system hydrogel) to embed the complex into the thermosensitive gel. However, the injection obtained by this method only achieves slow release within 12 hours. In addition, the cyclodextrin inclusion compound injection of progesterone Although it has been launched overseas, it is rapidly absorbed after intramuscular and subcutaneous injection, resulting in large fluctuations in blood concentration and the potential for adverse reactions (Cometti B. Pharmaceutical and clinical development of a novel progesterone for mulation [J]. Acta Obstet Gynecol Scand, 2015, 94 Suppl 161: 28-37.).

[0007] Furthermore, Chinese Patent Application Publication No. CN117064845A discloses a locally injectable, reversible, thermosensitive progesterone gel formulation for long-acting administration. This patent utilizes poloxamer as the gel matrix, polysorbate 80 as a solubilizer, polyethylene glycol 400 as a suspending agent, benzyl alcohol as an analgesic and preservative, and water as the dispersing solvent. This patent involves directly adding micronized progesterone to the gel matrix and then grinding it. The progesterone is suspended in the system as a crystal, similar to nanocrystal technology and a non-true solution formulation. This approach aligns with the technology employed in Chinese Patent Application Publication No. CN109223722A.

[0008] To achieve the slow release of progesterone, Chinese Patent Application Publication No. CN108635330A discloses a long-acting, sustained-release progesterone gel composition. The composition comprises water-insoluble phospholipids, cholesterol, a surfactant, and progesterone, which are dispersed into a uniform suspension via microfluidization to form a progesterone delivery solution. This delivery solution is then dispersed in a gel matrix of poloxamer 407. Because progesterone is poorly soluble in water, with a solubility of approximately 8 μg / mL, it is difficult to achieve clinical dosing concentrations using conventional surfactant solubilization. This invention is essentially a composite technology of liposomes and nanocrystals, using phospholipids to encapsulate a portion of the progesterone, surfactants to solubilize a portion, and microfluidization to convert another portion into tiny crystals under high pressure. This mixture is then dispersed in the gel matrix. Consequently, the progesterone in this formulation exists in both molecular and crystalline states, resulting in uncontrollable drug release during administration. The molecular state is released first, while the size of the crystal particles also affects the release rate. Therefore, the present invention is similar to nanocrystals in that progesterone is micronized by physical means and then suspended in a gel matrix, which is basically consistent with the technical route adopted in the patent document CN109223722A. However, the progesterone in the technical route adopted in the technical solution of the present invention exists in a molecular state and appears to be a clear liquid, which is essentially different from the technical route in the patent document CN108635330A.

[0009] To reduce the frequency of dosing, Chinese Patent Application Publication No. CN107441061A discloses a polyethylene glycol (PEG)-modified progesterone nanoparticle for injection. Progesterone and a polymer are dissolved in an organic solvent and then shear-emulsified with an aqueous solution containing an emulsifier to obtain a final emulsion. The solution is then cryogenically solidified, washed, concentrated, and finally lyophilized with a protective agent to produce sustained-release progesterone nanoparticles. The sustained-release progesterone injection containing these sustained-release progesterone nanoparticles has higher bioavailability than commercially available formulations and traditional progesterone nanoparticles, and achieves a sustained-release effect of more than one week. This prior art improves patient compliance with medication to a certain extent. However, since PEGylated drugs easily stimulate the body to produce anti-PEG antibodies after entering the body, thereby targetedly clearing substances containing PEG structures, the drug efficacy gradually decreases with prolonged use. This has been confirmed in long-circulating doxorubicin liposomes. Furthermore, the degradation mechanism in vivo is still unclear, and this solution still has potential nephrotoxicity in clinical application.

[0010] Based on the above, the development of a progesterone dosage form with good safety and long-term sustained-release effect is a technical problem that needs to be solved urgently in this field. Summary of the Invention

[0011] 1. Problem to be solved

[0012] The first object of the present invention is to provide a progesterone pharmaceutical composition with a long-lasting sustained-release effect and low irritation, which can greatly reduce the administration frequency of progesterone preparations and improve patient compliance.

[0013] The second object of the present invention is to further provide a progesterone composition with a reduced administration volume based on the above object, which can maintain a single administration dose at a lower volume to further improve patient compliance.

[0014] The third object of the present invention is to provide a progesterone injection with long-lasting sustained-release effect and low irritation.

[0015] The fourth object of the present invention is to provide a progesterone subcutaneous injection with a small single-dose volume, a long-lasting sustained-release effect, and low irritation.

[0016] 2. Technical solution

[0017] In order to solve the problem of high frequency of drug administration, a drug delivery system with a sustained-release effect can be used as a carrier to deliver progesterone. In situ gel is a very promising drug delivery system that can undergo phase change at the injection site and form a local drug delivery reservoir due to changes in the external environment (such as light, temperature, pH value and solvent exchange, etc.), with good tissue compatibility and the ability to release drugs for a long time at the administration site. The gel matrix of in situ gel generally adopts phospholipids, chitosan, poloxamer and polyethylene glycol block copolymers. Among them, phospholipids, as one of the components of the cell membrane, are easy to fuse with the cell membrane to deliver the drug into the cell to play a role, and have the best biocompatibility and safety. At the same time, the self-assembly characteristics of phospholipids also determine the property of phospholipids to self-assemble into vesicles with a bilayer structure when they come into contact with water. As an excellent drug reservoir, vesicles can store both fat-soluble drugs and water-soluble drugs. Therefore, the lipid in situ gel prepared with phospholipids as the main excipient can utilize the water in the tissue fluid to interact with phospholipids to form a multilayer vesicle structure after injection, thereby forming a drug reservoir and achieving a sustained release effect.

[0018] However, during the research process of the present invention, it was discovered that using only phospholipids as a sustained-release carrier for progesterone, due to their generally low phase transition temperature, the resulting vesicle bilayer is fluid and not dense, resulting in a relatively rapid drug release rate and unsatisfactory sustained-release effect. For example, in some embodiments, when the progesterone:phospholipid ratio is 1:4, the release rate is approximately 72.3% over 48 hours, based on which it is estimated that complete release should occur between 72 and 96 hours. While there is a certain sustained-release effect, the bilayer structure formed by the phospholipids is relatively weak, making it difficult to achieve a sustained release of progesterone for more than 144 hours.

[0019] To further extend the sustained release of progesterone from phospholipids, the addition of cholesterol to the sustained-release system is considered to enhance the strength of the phospholipid bilayer structure. Cholesterol and phospholipids are the building blocks of cell membranes, often forming a co-factor with phospholipids. Cholesterol can modulate the fluidity and permeability of the phospholipid bilayer, thereby effectively altering the drug release rate and preventing a burst effect caused by rapid drug release.

[0020] The addition of cholesterol to the sustained-release system achieved the purpose of prolonging the sustained-release time, but the surprising discovery was that the addition of cholesterol unexpectedly increased the drug loading concentration of progesterone.

[0021]

Drug composition

[0022] On this basis, the first aspect of the present invention provides a pharmaceutical composition comprising the following components:

[0023] a.Active ingredients;

[0024] b. sustained-release carrier;

[0025] c. Fat-soluble nonionic surfactant;

[0026] d. a pharmaceutically acceptable solvent;

[0027] Wherein, the active ingredient includes progesterone;

[0028] The sustained-release carrier includes phospholipids and cholesterol;

[0029] The pharmaceutically acceptable solvent includes a dispersing solvent;

[0030] The dispersing solvent includes NMP and a short-chain alcohol;

[0031] The active ingredient accounts for 5-13% (w / w) of the total weight of the pharmaceutical composition;

[0032] The ratio of the phospholipid to the active ingredient is (3.5-10):1 (w / w)

[0033] The ratio of cholesterol to active ingredient is (0.5-3.5):1 (w / w);

[0034] The ratio of the dispersing solvent to the active ingredient is (2-8):1 (w / w);

[0035] The ratio of the fat-soluble nonionic surfactant to the active ingredient is (0-1.2):1 (w / w);

[0036] The pharmaceutically acceptable solvent optionally includes a phase transition regulator, and the weight proportion of the phase transition regulator in the composition is 0-4%.

[0037] In the present invention, "optionally" means that the component may be present or not. When the weight ratio is 0, the component is not present. For example, in the above-mentioned composition, when the weight ratio of the fat-soluble nonionic surfactant and the phase transition modifier is 0, the component is not present.

[0038] In the compositions of the present invention, a sustained-release carrier refers to a carrier for the active ingredient, enabling the active ingredient to slowly release the drug as desired in a prescribed release medium after administration. Compared to conventional formulations without a sustained-release carrier, this reduces dosing frequency, significantly improves patient compliance, and in some cases, reduces drug side effects. Sustained-release carriers include phospholipids and cholesterol. Phospholipids are amphiphilic and can spontaneously self-assemble into lyotropic gels in the presence of water, thereby exerting a sustained-release effect.

[0039] As a preferred embodiment of any technical solution of the first aspect of the present invention, the ratio of the phospholipid to the active ingredient is (4-8):1 (w / w).

[0040] As a preferred embodiment of any technical solution of the first aspect of the present invention, in the composition of the present invention, the phospholipid is a natural phospholipid, including soybean lecithin, egg phospholipid or a combination thereof. Preferably, the content of phosphatidylcholine in the natural phospholipid is not less than 80%. The drug loading concentration in the liquid composition is affected by multiple factors, for example, the solubility of the active ingredient in the dispersing solvent, the volume of the dispersing solvent, etc. In the liquid composition, in order to obtain a uniform and stable system, the solubility of other non-liquid components such as sustained-release carriers in the dispersing solvent should also be considered. In the system of the first aspect of the present invention, the dispersing solvent is used to dissolve the active ingredient and the sustained-release carrier, and under the combined influence of NMP as a dispersing solvent and short-chain alcohols and cholesterol, on the one hand, the proportion of the dispersing solvent is reduced, and on the other hand, the solubility of progesterone is increased, and the drug loading concentration of progesterone can be ultimately increased to a maximum of 130 mg / mL. When the drug loading concentration is increased, in some application scenarios, such as subcutaneous injection, a reduction in the single injection dose can be achieved, thereby improving the patient's compliance with the medication from multiple perspectives such as convenience and irritation.

[0041] While increasing drug loading concentration, the addition of cholesterol effectively enhances the sustained release of progesterone compared to phospholipid systems. In vitro release results show that, within a certain range, increasing cholesterol dosage slows the initial release of progesterone, reducing burst release. Surprisingly, the addition of cholesterol also unexpectedly increases the drug loading concentration of progesterone in the system, further reducing the dosage volume and improving clinical compliance.

[0042] Based on the technical solution of the first aspect of the present invention, the applicant further surprisingly found that the promoting effect of cholesterol on the solubility of progesterone does not have a proportional relationship as expected. When the ratio of cholesterol to progesterone in the system exceeds a certain range, the drug-loaded concentration of progesterone in the system decreases.

[0043] Therefore, as a preferred embodiment of any technical solution of the first aspect of the present invention, the ratio of cholesterol to the active ingredient is (1-2.5):1 (w / w), preferably (1.5-2.5):1 (w / w). When the ratio of cholesterol to the active ingredient is less than 0.5:1, the promoting effect of cholesterol on the dissolution of progesterone in the system is not ideal. When the ratio is between 1:1 and 2.5:1, the promoting effect of cholesterol on the dissolution of progesterone in the system is very significant, and the solution remains a light yellow clear liquid after 60 days of storage; when the ratio of cholesterol to the active ingredient is greater than or equal to 4:1, the promoting effect of cholesterol on the dissolution of progesterone in the system disappears. Although a clear solution can be obtained, a small amount of fine crystals precipitate after the solution is placed for 24 hours; a large amount of fine crystals precipitate after the solution is placed for 60 days.

[0044] The selection of a dispersion medium is crucial for injectables. Dispersion media can dissolve raw materials and excipients, facilitating convenient administration. However, they can also be irritating. Therefore, the selection of a dispersion medium must balance solubility and irritation to ensure drug safety and efficacy. For long-acting injectables, cosolvents are often used in in-situ formulations. Cosolvents inhibit gel formation, reduce viscosity, and ensure injectability. Once the drug reaches the injection site, the cosolvent combines with the surrounding tissue fluid to form an in-situ gel. Upon interaction with the tissue fluid, the phospholipid-based in-situ gel undergoes a phase transition, forming a drug reservoir with bilayer characteristics. The speed of this phase transition depends on the miscibility of the dispersion medium with water. Higher miscibility results in a faster phase transition and faster drug reservoir formation, effectively minimizing drug burst release. Common cosolvents include NMP, ethanol, 1,2-propylene glycol, and glycerol. NMP is a water-soluble organic solvent with strong biocompatibility. Because NMP is water-soluble, it rapidly disperses with the tissue fluid in the tissue at the injection site after injection, allowing the formulation to form an in-situ gel. NMP not only has good water solubility but also has good solubility for progesterone, making it the preferred dispersing solvent in the present invention. However, other excipients, such as cholesterol and phospholipids, have limited solubility in NMP, necessitating the use of one or more co-solvents to address the solubility of these other excipients.

[0045] In the composition of the present invention, the short-chain alcohol in the dispersing solvent (dispersing medium) refers to a fatty alcohol having no more than 3 carbon atoms, including a linear or branched saturated or unsaturated fatty alcohol, and the fatty alcohol can be a monohydric alcohol or a polyhydric alcohol, such as ethanol, propanol, propylene glycol, or glycerol.

[0046] As a preferred embodiment of any technical solution of the first aspect of the present invention, the short-chain alcohol includes one or more of ethanol, propylene glycol or glycerol.

[0047] As a preferred embodiment of any technical solution of the first aspect of the present invention, the short-chain alcohol comprises at least ethanol. Ethanol is a commonly used solvent in the preparation, has good biosafety, and also has good diffusivity, which can accelerate the speed at which the preparation forms a gel when it encounters an aqueous phase. The solubility of the phospholipid in the present invention in ethanol is greater than its solubility in NMP. Therefore, the present invention selects NMP and ethanol as co-solvents to solve the solubility problem of progesterone and excipients (phospholipids, cholesterol, surfactant).

[0048] The toxicity of the solvent determines its maximum dosage in the preparation. In order to further improve the safety of the preparation and reduce the impact of the solvent on the injection site after injection, the present invention screens the solvent dosage based on the safety of the composition and the drug loading concentration to obtain a solvent dosage suitable for the composition of the present invention.

[0049] As a preferred embodiment of any technical solution of the first aspect of the present invention, the dispersing solvent contains NMP and ethanol, and the ratio of NMP to ethanol is (2-4):1 (w / w).

[0050] As a preferred embodiment of any technical solution of the first aspect of the present invention, the ratio of the dispersing solvent to the active ingredient is (2-8):1 (w / w), preferably (3-6):1 (w / w).

[0051] In order to further increase the drug loading concentration and reduce the irritation caused by the dispersing solvent, based on any technical solution of the first aspect of the present invention, another object of the present invention is to further reduce the proportion of the dispersing solvent in the composition.

[0052] As a preferred embodiment of any technical solution of the first aspect of the present invention, a surfactant is added to the pharmaceutical composition to reduce the amount of the dispersing solvent while ensuring the drug loading concentration. In essence, the addition of a surfactant reduces the amount of the dispersing solvent. On the one hand, while ensuring the drug loading, the overall volume of the composition is reduced, thereby achieving an increase in the actual drug loading mass volume concentration. During the research process of the present invention, with the addition of a surfactant, the progesterone drug loading in the lipid gel constructed of phospholipids and cholesterol is effectively increased. On the other hand, the reduction in the amount of dispersing solvent can simultaneously reduce the irritation during administration. In some application scenarios, such as subcutaneous injection or intramuscular injection, it can reduce symptoms such as local redness, pain, and nodules at the injection site.

[0053] As a preferred embodiment of any technical solution of the first aspect of the present invention, the fat-soluble nonionic surfactant is GMS.

[0054] Since GMS itself is a solid, it needs to be dissolved in the dispersion solvent before enhancing the dissolution of progesterone. Therefore, its addition amount is also limited by the amount of dispersion solvent. When the GMS addition ratio is too high, although the amount of dispersion solvent can be further reduced, the stability of the composition is deteriorated, and the system is prone to crystal precipitation after being left for a period of time after dissolution. Therefore, as a preferred embodiment of any technical solution of the first aspect of the present invention, the ratio of GMS to active ingredient is (0-1.2):1 (w / w), more preferably (0-1):1 (w / w).

[0055] Another unexpected discovery is that, compared with the composition without GMS, the addition of GMS as a surfactant further prolongs the sustained-release time of the active ingredient while meeting the stability requirements of the composition.

[0056] As a preferred embodiment of any technical solution of the first aspect of the present invention, the pharmaceutical composition comprises the following components:

[0057] a.Active ingredients;

[0058] b. sustained-release carrier;

[0059] c. Fat-soluble nonionic surfactant;

[0060] d. a pharmaceutically acceptable solvent;

[0061] Wherein, the active ingredient includes progesterone;

[0062] The sustained-release carrier includes phospholipids and cholesterol;

[0063] The pharmaceutically acceptable solvent includes a dispersing solvent;

[0064] The dispersing solvent includes an organic solvent NMP and a short-chain alcohol;

[0065] The active ingredient accounts for 5-13% (w / w) of the total weight of the pharmaceutical composition;

[0066] The ratio of the phospholipid to the active ingredient is (4-8):1 (w / w);

[0067] The ratio of cholesterol to active ingredient is (1-2.5):1 (w / w);

[0068] The ratio of the dispersing solvent to the active ingredient is (3-6):1 (w / w);

[0069] The ratio of the fat-soluble nonionic surfactant to the active ingredient is (0-1):1 (w / w).

[0070] As a preferred embodiment of any technical solution of the first aspect of the present invention, the phase transition modifier includes water. Preferably, the water is water for injection.

[0071] The injection water can adjust the phase change time of the in situ gel preparation formed by the pharmaceutical composition. In order to maximize the speed of phase change after the in situ gel is injected, a certain proportion of injection water can be added to the present invention.

[0072] As a preferred embodiment of any technical solution of the first aspect of the present invention, the water accounts for 0-4% of the total weight of the composition. Within this range, the pharmaceutical composition does not undergo phase transition during storage and is suitable for use as an injection.

[0073] As a preferred embodiment of any technical solution of the first aspect of the present invention, the water accounts for 4%-7% of the total weight of the composition, excluding 4%. Within this percentage range, the pharmaceutical composition undergoes phase transition and is suitable for use as a gel.

[0074] As a preferred embodiment of any technical solution of the first aspect of the present invention, the pharmaceutical composition is a drug for administration by injection, or a drug for administration by subcutaneous implantation, or a drug for external use and vaginal administration.

[0075] According to a preferred embodiment of any of the technical solutions of the first aspect of the present invention, the form (dosage form) of the pharmaceutical composition having a preventive effect and / or the pharmaceutical composition having a therapeutic effect is mainly a liquid form, which can be directly formulated or prepared into a form of raw materials that can be used for pharmaceuticals, quasi-drugs, etc. Preferably, the dosage form of the pharmaceutical composition includes a sustained-release formulation, a controlled-release formulation, a pulsed-release formulation, and an injection. The formulation and preparation methods of the pharmaceutical composition of the present invention in these pharmaceutical formulation forms can be achieved by methods and experience known to those skilled in the art.

[0076] As a preferred embodiment of any technical solution of the first aspect of the present invention, the pharmaceutical composition is in the form of an injection.

[0077] As a preferred embodiment of any technical solution of the first aspect of the present invention, the injection solution includes subcutaneous injection solution or intramuscular injection solution.

[0078] As a preferred embodiment of any technical solution of the first aspect of the present invention, the mass percentage of progesterone in the pharmaceutical composition is selected from any value within the following group of numerical ranges:

[0079] 5.0~12.0wt%;

[0080] 5.0~11.0wt%;

[0081] 5.0~10.0wt%, 5.5~10.0wt%, 6.0~10.0wt%, 6.5~10.0wt%, 7.0~10.0wt%, 7.5~10.0wt%, 8.0~10.0wt%, 8.5~10.0wt%, 9.0~10.0wt%, 9.5~10.0wt%;

[0082] 5.0~9.0wt%, 5.5~9.0wt%, 6.0~9.0wt%, 6.5~9.0wt%, 7.0~9.0wt%, 7.5~9.0wt%, 8.0~9.0wt%, 8.5~9.0wt%;

[0083] 5.0~8.0wt%, 5.5~8.0wt%, 6.0~8.0wt%, 6.5~8.0wt%, 7.0~8.0wt%, 7.5~8.0wt%.

[0084] It is preferably 5.5 to 10.0 wt %; more preferably 6.0 to 9.0 wt %; and most preferably 7.5 to 8.5 wt %.

[0085] As a preferred embodiment of any technical solution of the first aspect of the present invention, the concentration of progesterone in the pharmaceutical composition (drug loading concentration) is 55 to 130 mg / mL.

[0086] It is preferably 55 to 100 mg / mL; preferably 60 to 100 mg / mL; and more preferably 60 to 80 mg / mL.

[0087] Injection

[0088] The second aspect of the present invention provides an injection comprising the pharmaceutical composition described in any technical solution of the first aspect of the present invention.

[0089] According to the injection described in any technical solution of the second aspect of the present invention, the injection is a subcutaneous injection or an intramuscular injection.

[0090] In situ gel

[0091] The third aspect of the present invention provides an in situ gel comprising the pharmaceutical composition described in any technical solution of the first aspect of the present invention.

[0092]

Sustained-release depot drugs

[0093] The fourth aspect of the present invention provides a sustained-release depot drug comprising the pharmaceutical composition according to any one of the technical solutions in the first aspect of the present invention. It should be noted that the in situ gel can be regarded as a specific dosage form of the sustained-release depot drug.

[0094] Preparation method of pharmaceutical composition

[0095] The fifth aspect of the present invention provides a method for preparing the pharmaceutical composition according to any technical solution of the first aspect of the present invention, comprising the following steps:

[0096] Preparing a sustained-release carrier solution: dissolving the sustained-release carrier in a pharmaceutically acceptable solvent;

[0097] Dissolving the active ingredient: dissolving the active ingredient in a sustained-release carrier solution to obtain a pharmaceutical composition.

[0098] As a preferred embodiment of any technical solution of the fifth aspect of the present invention, before dissolving the active ingredient in the sustained-release carrier solution, a surfactant is dissolved in the sustained-release carrier solution.

[0099] As a preferred embodiment of any technical solution of the fifth aspect of the present invention, the solution obtained after dissolving the active ingredient is filtered using a filter membrane; the filter membrane is preferably a filter membrane with a pore size of no more than 0.22 μm.

[0100] As a preferred embodiment of any technical solution of the fifth aspect of the present invention, water for injection is added to the solution obtained after dissolving the active ingredient. Preferably, the water for injection is filtered using a filter membrane; the filter membrane preferably has a pore size of no greater than 0.22 μm.

[0101] As a preferred embodiment of any technical solution of the fifth aspect of the present invention, the pharmaceutical composition is stored in a sterile vial. Preferably, the pharmaceutical composition is stored in a light-proof environment at a storage temperature not higher than 30°C.

[0102] As a preferred embodiment of any technical solution of the fifth aspect of the present invention, the pharmaceutical composition can be sterilized by a moist heat sterilization process.

[0103] Specifically, the preparation method of the pharmaceutical composition includes:

[0104] (1) Weigh the prescribed amount of phospholipid, add the prescribed amount of anhydrous ethanol, stir or ultrasonically dissolve, add NMP after dissolution, and mix well;

[0105] (2) Add the prescribed amount of cholesterol. If the formula contains GMS, add the prescribed amount of GMS and stir or ultrasonically dissolve;

[0106] (3) Add the prescribed amount of active ingredient and dissolve;

[0107] (4) Filter through a 0.22 μm filter membrane. If the formula contains water, add the prescribed amount of 0.22 μm filtered water for injection to the filtered sample and stir evenly.

[0108] (5) Dispense into sterile vials, fill with nitrogen, stopper, cap, and store at room temperature, away from light.

[0109] (6) The capped samples can also be sterilized by a wet heat sterilization process at 121°C / 12 min, and the finished products can be stored away from light after sterilization.

[0110] Pharmaceutical applications of drug compositions, injections, in-situ gels, and sustained-release reservoir drugs

[0111] The sixth aspect of the present invention provides the pharmaceutical composition of the first aspect of the present invention, or the injection of the second aspect of the present invention, or the in situ gel of the third aspect of the present invention, or the sustained-release reservoir drug of the fourth aspect of the present invention, and the use of any of these technical solutions in the preparation of drugs for preventing or treating menstrual disorders, such as amenorrhea and functional uterine bleeding, luteal insufficiency, threatened abortion and habitual abortion.

[0112] As a preferred embodiment of any technical solution of the sixth aspect of the present invention, the concentration of progesterone in the pharmaceutical composition is 55-130 mg / mL.

[0113] It is preferably 55 to 100 mg / mL; preferably 60 to 100 mg / mL; and more preferably 60 to 80 mg / mL.

[0114] When administered at the above drug loading concentration, satisfactory sustained-release effects can be obtained.

[0115] Any embodiment (technical solution) of any aspect of the present invention may be combined with other embodiments, provided that no contradiction arises. In addition, in any embodiment of any aspect of the present invention, any technical feature may be applicable to the technical feature in other embodiments, provided that no contradiction arises.

[0116] Provided that no contradiction arises, any technical feature of any aspect of the present invention or any embodiment of such aspect is also applicable to any other embodiment or any embodiment of any other aspect. Of course, when applicable to each other, the corresponding features may be appropriately modified as necessary. The various aspects and features of the present invention are further described below.

[0117] In addition, the various terms and phrases used in the present invention have general meanings known to those skilled in the art. Even so, the present invention still hopes to provide a more detailed description and explanation of these terms and phrases herein. If the mentioned terms and phrases are inconsistent with the known meanings, the meanings expressed in the present invention shall prevail.

[0118] In the present invention, the term "comprising" or "containing" means that various components can be used together in the composition of the present invention.

[0119] The composition of the present invention also includes isomers, solvates, or pharmaceutically acceptable salts thereof of the specific compounds in the above composition, as long as they have the same or substantially the same function as the specific compounds in the composition.

[0120] The term "in situ gel" in the present invention refers to a preparation in which progesterone is administered in a solution state and undergoes a phase transition at the administration site, solidifying from a liquid to form a semi-solid gel.

[0121] The term "sustained-release reservoir drug" in the present invention refers to an injection solution with progesterone as the active ingredient and phospholipids, cholesterol, GMS, NMP and ethanol as components, which forms a drug reservoir at the injection site and slowly releases the drug to produce the drug effect.

[0122] The term "phase transition regulator" in the present invention refers to water, which refers to a substance that can accelerate the speed at which the composition changes from a clear liquid to a light yellow or white semi-solid gel after being added.

[0123] The actual dosage level and route of administration of the active ingredients in the pharmaceutical compositions of the present invention may be varied, for example, by increasing the drug loading concentration, increasing the volume of administration, or increasing the frequency of administration, so as to obtain an amount of active substance effective to achieve the desired therapeutic response in a particular patient. The dosage level should be selected based on the activity of the specific active substance, the route of administration, the severity of the condition being treated, and the condition and medical history of the patient being treated.

[0124] Therefore, compared with the prior art, the present invention uses phospholipids and cholesterol as the main excipients to construct a progesterone pharmaceutical composition, effectively increasing the drug loading concentration of progesterone and achieving a slow release of progesterone. In vivo experiments show that the sustained release time of the pharmaceutical composition 3 in the embodiment reaches more than 168 hours. In particular, when the pharmaceutical composition is administered as a progesterone injection for subcutaneous injection, due to the high drug loading concentration, the injection can form a drug reservoir after injection at the lowest possible injection volume, achieving the dual purpose of long-term release and reducing irritation. It is an ideal long-acting preparation with high drug loading, long release time, low injection irritation, and good patient compliance.

[0125] 3. Beneficial effects

[0126] Compared with the prior art, the present invention has the following beneficial effects:

[0127] (1) The design process of the present invention has always been centered around the dissolution and release of progesterone. Through a large number of experimental studies, a progesterone pharmaceutical composition was invented. The pharmaceutical composition can be used as a lipid in situ gel, has a long-lasting sustained-release effect, and significantly reduces the frequency of administration and injection irritation. Furthermore, in Example 14, after pharmacokinetic testing, the duration of pharmaceutical composition 3 in rabbits can reach more than 168 hours. In the rabbit endometrial conversion pharmacodynamics experiment, the progesterone pharmaceutical composition in the test group was only administered once a week, repeated 3 times, while the progesterone oil injection in the control group was administered continuously for 21 days. The experimental results surprisingly found that the progesterone pharmaceutical composition in the test group effectively increased the concentration of progesterone in the uterus. The progesterone concentration in the test group was about 15 times that of the control group, and the uterine coefficient was nearly 2 times that of the control group. The results of uterine staining sections showed that the number of glands in the progesterone pharmaceutical composition in the test group was significantly more than that in the control group. Furthermore, in Example 11, the progesterone pharmaceutical composition showed no irritation at the injection site compared to the progesterone oil injection. However, the progesterone oil injection group developed erythema at the injection site seven days after administration, indicating mild irritation. Therefore, compared to the prior art, the pharmaceutical composition prepared by the present invention exhibits a long-lasting sustained-release effect, exerts superior efficacy, and significantly reduces dosing frequency and injection irritation. This will significantly improve patient compliance during clinical use and effectively reduce treatment procedures.

[0128] (2) Based on the purpose of long-term sustained release, the present invention adds cholesterol to the composition. Cholesterol can change the fluidity of the membrane formed after the phase transition of the phospholipid reservoir, making the phospholipid membrane after the phase transition more stable, effectively controlling the sudden release of the drug, and exerting a long-term effect together with the phospholipid. However, the present invention surprisingly found that the addition of cholesterol to the composition effectively increased the concentration of the drug. For example, on the basis of a prescription without cholesterol, a certain amount of cholesterol was added so that the weight ratio of cholesterol to progesterone was 1:1, and the drug loading concentration of the pharmaceutical composition increased from about 68 mg / ml to about 116 mg / ml. At the same time, the increase in drug loading concentration promoted by cholesterol can only be achieved under a specific ratio, and the increase in drug loading concentration significantly reduces the administration volume or administration frequency of the pharmaceutical composition preparation of the present invention, thereby improving the convenience of clinical use of the preparation.

[0129] (3) The present invention further discovered that the fat-soluble nonionic surfactant GMS unexpectedly exhibited the effect of delaying drug release in the composition. As shown in the data of Example 8, the cumulative release rate of the composition without GMS in 72 hours was about 58.3%, while the cumulative release rate of the composition after adding GMS in 72 hours was about 39.8%, indicating that it further significantly reduced the drug release rate of the phospholipid reservoir, which can make the composition have a longer release time, effectively achieving the invention purpose of long-term sustained release. At the same time, the present invention uses GMS as a fat-soluble nonionic surfactant to effectively increase the concentration of the drug in the composition, thereby further reducing the dosage volume or dosage frequency of the preparation, which is more conducive to improving the convenience of clinical use of the preparation.

[0130] (4) Further compositions of the present invention may contain a small amount of water. Water, as a phase transition modifier, can effectively regulate reservoir formation. For example, in the in vitro experiment of Example 12, when approximately 7% of water was added to the system, the composition transformed from a clear liquid to a milky white semisolid, compared to the composition without water. Therefore, the addition of water to the system can accelerate the formation of an in situ gel at the injection site and reduce burst release to a certain extent. BRIEF DESCRIPTION OF THE DRAWINGS

[0131] FIG1 shows the appearance of pharmaceutical composition 3 in Example 12 and its appearance after phase change.

[0132] Figure 2 shows the safety and in vivo development of the solvent group and composition 1 in Example 13 24 hours and 168 hours after injection, respectively (the right figure is a partial enlarged view of the left figure).

[0133] FIG3 is a diagram showing the irritation at the injection site and the in vivo formation in the rabbit endometrial transformation experiment in Example 14.

[0134] FIG4 shows the uterine weight (A) and the progesterone concentration in the uterine tissue (B) in the rabbit endometrial transformation experiment in Example 14.

[0135] FIG5 is a pathological section of the uterus in the rabbit endometrial transformation experiment in Example 14. DETAILED DESCRIPTION

[0136] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains; the term "and / or" used herein includes any and all combinations of one or more of the associated listed items.

[0137] If the specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or those recommended by the manufacturer. All reagents or instruments used, if the manufacturer is not specified, are commercially available conventional products.

[0138] As used herein, the term "about" is used to provide flexibility and imprecision associated with a given term, measurement, or value. One skilled in the art can readily determine the degree of flexibility for a particular variable.

[0139] Concentration, amount and other numerical data can be presented in range format in this article.Should be understood that such range format is only used for convenience and brevity, and should be flexibly interpreted as not only including the numerical value clearly described as range limit, but also including all independent numerical values ​​or subranges encompassed within the scope, just as each numerical value and subrange are clearly described.For example, the numerical range of about 1 to about 4.5 should be interpreted as not only including the limit value of 1 to about 4.5 clearly described, but also including independent numerals (such as 2,3,4) and subranges (such as 1 to 3,2 to 4 etc.).The same principle is applicable to the scope of only narrating a numerical value, such as "less than about 4.5", which should be interpreted as including all above-mentioned values ​​and scopes.In addition, no matter how the breadth of described scope or feature is, this explanation should be applicable.

[0140] The present invention will be further described below with reference to specific embodiments.

[0141] Example 1

[0142] Study on the solubility of active ingredients and phospholipids in solvents

[0143] Combining the solubility of the drug in the solvent and the solubilization effect of the surfactant, a combined approach was used to improve the solubility of the target drug, and the solvent or solvent combination was used to screen for the optimal system for dissolving progesterone. The results are shown in Table 1.

[0144] Table 1 Solubility of progesterone in different solvents or solvent combinations and rat toxicity data of the solvents

[0145] As can be seen from the above table, among the single solvents, progesterone has the highest solubility in NMP, followed by ethanol. In addition, animal toxicity data show that each dispersion medium has good biosafety when used for subcutaneous injection. According to the dosage calculation of the present invention, the amount of each dispersion solvent used for injection is lower than LD 50 Therefore, using NMP and ethanol as dispersion solvents has good safety.

[0146] Phospholipids have the property of self-assembly, spontaneously forming vesicle structures in water, effectively encapsulating active ingredients. As shown in the table above, the addition of phospholipids can improve the solubility of active ingredients in ethanol, and the type and amount of phospholipids directly affect their solubility in the solvent. To further screen for suitable phospholipids, solubility experiments were conducted on phospholipids of different types and with varying phosphatidylcholine (PC) content. 0.5g of phospholipid and 1g of solvent (ethanol or NMP) were placed in a sealed container and placed in a 50°C water bath. The dissolution time and state of the different phospholipids were observed. The results are shown in Table 2.

[0147] Table 2 Phospholipid solubility experiment

[0148] According to the above results, ethanol is more suitable as a good solvent for phospholipids than NMP. Natural soybean lecithin and egg yolk lecithin have good solubility in ethanol, while synthetic dipalmitoylphosphatidylcholine (DPPC) and hydrogenated soybean lecithin (HSPC) cannot be completely dissolved in ethanol. As the PC content in soybean lecithin increases, its dissolution rate in ethanol is faster. Based on this, the present invention prefers soybean lecithin with a purity of 95% or 80% egg yolk lecithin as a sustained-release carrier.

[0149] In summary, solubility studies show that the target active ingredient, progesterone, has the highest solubility in NMP, followed by ethanol. Phospholipids, as sustained-release carriers, have the highest solubility in ethanol. Therefore, NMP and ethanol are preferred co-solvents for dissolving the target active ingredient and phospholipids.

[0150] Example 2

[0151] Screening of phospholipid ratio

[0152] A sample containing progesterone was prepared according to the prescription in Table 3, and the dissolution of the progesterone was observed at room temperature.

[0153] Table 3 Prescription composition and solution properties

[0154] Experimental results show that phospholipids have a certain solubilizing effect on the fat-soluble drug progesterone due to their amphiphilic nature. Therefore, the preferred ratio of progesterone to phospholipids in the present invention is 1:(3.5-8), and more preferably 1:(4-8).

[0155] Example 3

[0156] Screening of the ratio of dispersing solvent NMP to ethanol

[0157] The samples were prepared according to the prescription in Table 4, and the dissolution of progesterone was observed at room temperature.

[0158] Table 4 Prescription composition and dissolution

[0159] Experimental results show that when the ratio of NMP to anhydrous ethanol is (2-4):1, the sample is a yellow clear liquid, so this ratio range is preferred for the prescription of the present invention.

[0160] Example 4

[0161] Cholesterol ratio screening

[0162] In order to explore the effect of cholesterol on the solubility of progesterone, samples containing progesterone were prepared according to the prescription in Table 5, placed at room temperature for 24 h and 60 d, and the properties were observed.

[0163] Table 5 Prescription composition and properties

[0164] In the above formula, the progesterone:cholesterol ratio of 1:4 precipitated crystals. No crystals were precipitated in the other groups after 60 days. Therefore, the preferred ratio of progesterone to cholesterol in the present invention is 1:(1-2.5).

[0165] Example 5

[0166] Screening of surfactant types and optimization of content

[0167] The addition of surfactants helps increase the solubility of a substance in a solvent and is a common method for solubilizing pharmaceutical formulations. To further enhance the solubility of progesterone, samples were prepared as shown in Table 6 to investigate changes in the solubility of progesterone.

[0168] Table 6 Prescription composition and dissolution

[0169] The experimental results show that GMS, as a fat-soluble nonionic surfactant, can effectively dissolve progesterone under the condition of reducing solvent after addition.

[0170] The changes in the solubility of progesterone in the system after adding different proportions of GMS were investigated. Samples containing progesterone were prepared according to Table 7 and placed at room temperature for 24 hours, 60 days, and at a constant temperature (25°C) for 60 days to investigate the solubility of progesterone.

[0171] Table 7 Prescription composition and dissolution

[0172] The above samples were placed in a sample room at room temperature and 25°C for 60 days and observed. The results showed that no crystals were precipitated when the progesterone:GMS ratio was 1:0 and 1:1, proving that the system was clear, stable and uniform within this range. Therefore, the preferred weight ratio of progesterone to GMS in the present invention is 1:(0-1.2), and more preferably 1:(0-1).

[0173] Example 6

[0174] Effect of phospholipid ratio on sustained-release effect

[0175] Samples containing different proportions of phospholipids were prepared according to the formulation in Table 8, and the in vitro release was investigated.

[0176] In vitro release experiment: About 1.0 g of the composition sample was taken and placed in a dialysis bag (3000 Da). The dialysis bag was placed in 250 mL of phosphate buffer (pH 7.4) and kept at 37°C with a stirring speed of 100 rpm. Samples were taken at different time points and measured.

[0177] Table 8 Prescription composition and cumulative release data

[0178] Experimental results showed that the higher the phospholipid content in the formulation, the slower the release of progesterone from the sample, demonstrating that increasing the phospholipid content can effectively delay the release of progesterone. However, as the proportion of phospholipids in the sample increases, the solution viscosity increases, which may cause the progesterone to be too tightly packaged, slowing the release rate and causing the released concentration in the body to be lower than the effective concentration.

[0179] Example 7

[0180] Effect of cholesterol ratio on sustained-release effect

[0181] Samples were prepared according to the formulation in Table 9, and the cumulative release data were determined according to the method in Example 6.

[0182] Table 9 Formulation composition and cumulative release data

[0183] The results surprisingly revealed that the addition of cholesterol slowed the release rate of the composition compared to a cholesterol-free formulation. Furthermore, increasing cholesterol levels effectively reduced the burst release of the active ingredient. Cholesterol within this range can alter the fluidity and integrity of the phospholipid layer, improving stability and preventing rapid release of the active ingredient within the first four hours.

[0184] Example 8

[0185] Effect of GMS ratio on sustained-release effect

[0186] It is surprisingly found that, compared with the composition without GMS, the addition of GMS as a surfactant further prolongs the sustained-release time of the active ingredient while meeting the stability requirements of the composition.

[0187] Samples were prepared according to the formulation in Table 10, and the cumulative release data were determined according to the method in Example 6.

[0188] Table 10 Formulation composition and cumulative release data

[0189] The results showed that the addition of GMS could slow down the release rate of progesterone. In summary, the addition of GMS to the lipid in situ gel not only reduced the amount of solvent used but also increased the solubility of progesterone and slowed the release rate of progesterone.

[0190] Example 9

[0191] In order to further verify the proportion of water in the total weight of the composition, samples were prepared according to the formulation in Table 11, placed at room temperature for 24 hours, and the properties were observed.

[0192] Table 11 Prescription composition and properties

[0193] The results showed that the addition of water can alter the state of the system. When the amount of water added is too high, the system becomes unstable, manifested by the solution changing from clear to turbid. This experiment simulated the in vitro phenomenon of the in situ gel being injected into the body, demonstrating that it utilizes the interaction between water in the tissue fluid and phospholipids to form an in situ gel-type sustained-release drug reservoir at the injection site.

[0194] Example 10

[0195] In order to further verify the effect of the composition of the present invention, a pharmacokinetic study in rats was performed.

[0196] Composition 1 and Composition 2 were prepared according to the prescription in Table 12. Composition 1 had a progesterone concentration of 66 mg / mL, or 7.04% by weight, and Composition 2 had a progesterone concentration of 66 mg / mL, or 6.67% by weight. The preparation method for Composition 1 or Composition 2 is as follows:

[0197] (1) Weigh the prescribed amount of phospholipid, add the prescribed amount of anhydrous ethanol, stir to dissolve, and then add NMP after dissolution;

[0198] (2) Add the prescribed amount of cholesterol and active ingredient and dissolve;

[0199] (3) Filter using a 0.22 μm filter membrane;

[0200] (4) Dispense into sterile vials, stopper and place in a dark place.

[0201] The storage temperature should be below 30℃.

[0202] Table 12 Prescription composition of composition 1 and composition 2

[0203] The experimental animals were male SD rats, randomly divided into three groups and administered subcutaneously to the following regimens. Production License Number: SCXK(Zhejiang)2019-0001; Certificate Number: 20230106Aazz0619000180.

[0204] Table 13 Dosage regimen

[0205] *20 mg / mL, Zhejiang Xianjun Pharmaceutical Co., Ltd.

[0206] The experiment was conducted using a commercially available progesterone injection (20 mg / mL, also known as progesterone oil injection) as a control. The dosing schedule is shown in Table 14. Blood samples were collected from the retinal venous plexus at 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after administration for content testing. The results are shown in Tables 14-16:

[0207] Table 14 Concentration-time data of progesterone in plasma after subcutaneous injection of progesterone oil injection (ng / mL)

[0208] Note: BLQ: below the limit of detection.

[0209] Table 15 Concentration-time data of progesterone in plasma after subcutaneous injection of composition 1 (ng / mL)

[0210] Table 16 Concentration-time data of progesterone in plasma after subcutaneous injection of composition 2 (ng / mL)

[0211] The processed data are shown in Tables 14-16:

[0212] Table 17 Pharmacokinetic parameters of progesterone in plasma after subcutaneous administration of progesterone injection

[0213] Table 18 Pharmacokinetic parameters of progesterone in plasma after subcutaneous administration of Composition 1

[0214] Table 19 Pharmacokinetic parameters of progesterone in plasma after subcutaneous administration of composition 2

[0215] After subcutaneous injection of progesterone oil injection, the progesterone in the rat plasma was basically completely eliminated about 72 hours after administration; after subcutaneous injection of composition 1 and composition 2, the concentration of progesterone in the rat plasma was still detectable until 120 hours after administration, and remained at a relatively stable level from 24 hours to 96 hours after administration. The half-life was significantly improved compared with that of progesterone oil injection, indicating that a better sustained-release effect was achieved.

[0216] Composition 1 and composition 2 compared with the control group progesterone oil injection t 1 / 2 The results showed that the release rate of C max The release rates were only 29.4% and 34.8% of the control group, respectively. Compared with the control group, the release rate was mild and there was no burst release. The AUCs were 58.4% and 63.3% of the control group, respectively. This may be because the detection time was cut off at 120 hours and the measurement was not complete.

[0217] Example 11

[0218] The experimental animals were male beagle dogs (source: Animal Center, Medical College of Southeast University). Each beagle dog was subcutaneously injected with Composition 3 in Table 20. The progesterone concentration of Composition 3 was 66 mg / mL, the mass percentage was 6.06%, the dosage was 2 mg / kg, and the administration was only one injection.

[0219] The preparation method of composition 3 is:

[0220] (1) Weigh the prescribed amount of phospholipid, add the prescribed amount of anhydrous ethanol, stir to dissolve, and then add NMP after dissolution;

[0221] (2) Add the prescribed amount of cholesterol and GMS and dissolve by stirring or ultrasonication;

[0222] (3) Add the prescribed amount of active ingredient and dissolve;

[0223] (4) Filter using a 0.22 μm filter membrane;

[0224] (5) Dispense into sterile vials, stopper and place in a dark place.

[0225] The storage temperature should be below 30℃.

[0226] Table 20 Prescription composition of composition 3

[0227] Blood samples were collected from the cephalic vein of the forelimb at 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 24 hours, 32 hours, 48 ​​hours, 72 hours, 96 hours, 120 hours, and 144 hours after administration for content testing. The data are shown in Table 21:

[0228] Table 21 Concentration-time data of progesterone in plasma after subcutaneous injection of composition 3 (ng / mL)

[0229] The processed data are shown in Tables 22-23.

[0230] Table 22 Pharmacokinetic parameters of progesterone in beagle dog plasma

[0231] As can be seen from Table 22, compared with the reference progesterone oil injection (intramuscular injection), the average MRT of progesterone composition 3 is 41.55h, while the average MRT of progesterone oil injection recorded in the literature is 14.69h (Rui Li, Chen Ying, Chen Xun, Zhang Ziqiang. Pharmacokinetics study of progesterone preparations in Beagledog [J]. Journal of Pharmaceutical Research, 2022, 41: 709-712.); the elimination half-life of composition 3 is 64.34h, while the elimination half-life of progesterone oil injection is only 12.11h, indicating that composition 3 has a longer retention time in the body and has a longer pharmacological effect. Compared with progesterone oil injection, the C of composition 3 is 14.69h. max Decrease, T max The prolonged release further proves that the composition of the present invention has a sustained release effect.

[0232] Table 23 Drug concentration at different time points accounted for C max proportion

[0233] Table 23 shows the blood drug concentrations at different collection points and C max The ratio of C further proves the sustained release effect of the composition. The drug concentration can still be monitored at 144h, and its intensity is about C max The sustained-release effect can reach more than 144h.

[0234] Example 12

[0235] Application of pharmaceutical composition as in situ gel or sustained-release reservoir drug

[0236] FIG1 shows the changes in the appearance of the sample before and after a certain amount of water (water accounts for about 7% of the total weight of the composition) is added to the pharmaceutical composition 3 in Example 11.

[0237] The above experiments simulated the changes in the properties of the pharmaceutical composition after phase transition during the exchange of interstitial fluid and the pharmaceutical composition in the body during subcutaneous injection. The above results can further show that the pharmaceutical composition of the present invention can form a gel after interacting with interstitial fluid at the injection site, becoming a sustained-release depot drug and slowly releasing the active ingredient.

[0238] Example 13

[0239] To further prove the safety of the pharmaceutical composition, 4 rats, half male and half female, were taken and divided into two groups. One group was subcutaneously injected with 1 mL of solvent (NMP: absolute ethanol = 2:1), and the other group was subcutaneously injected with 1 mL of Composition 1 in Example 10. The animals were dissected at 24 h and 168 h after injection respectively to observe whether there was bleeding and color change of tissues subcutaneously, and at the same time, the changes in the administration group were observed.

[0240] The results showed that there were no redness or swelling phenomena in the subcutaneous tissues of the rats after 24 h and 168 h of administration of the solvent and the composition, indicating that both the composition and the solvent were non-irritating at this dose. The results are shown in Figure 2.

[0241] Example 14

[0242] Pharmacodynamic data

[0243] In this example, the "progesterone pharmaceutical composition group", "pharmaceutical composition 3" or "composition 3" all refer to the composition formula in pharmaceutical composition 3 in Example 11.

[0244] Through the rabbit endometrial transformation experiment, the biological activity of the progesterone pharmaceutical composition was further verified. The experimental animals were 15 immature New Zealand female rabbits, 6 weeks old, with a body weight of 0.5 - 0.8 kg. Original source: Hubei Yizhicheng Biotechnology Co., Ltd., production license number: SCXK(E)2021 - 0020, certificate number: 42817300002822.

[0245] The progesterone oil injection group was injected intramuscularly into the hind legs at 5 mg / kg, with both legs administered alternately once a day for a total of 21 times. The progesterone pharmaceutical composition group was Composition 3 of the pharmaceutical composition, administered subcutaneously at the neck at 35 mg / kg, with an injection frequency of once a week for a total of 3 times. The solvent group was the pharmaceutical composition 3 without active ingredients, with the same injection site, dose and frequency as the pharmaceutical composition 3 group. At 1, 3, 7, 14 and 21 days after administration, the irritation at the injection site of the solvent group, the pharmaceutical composition 3 group and the progesterone oil injection group was observed, and the injection site was photographed and scored. After 21 days, the rabbits were euthanized and the uterus was taken. The wet weights of the 2 uteri of each female rabbit were weighed. One was stored at 4°C for the detection of drug tissue distribution; the other was fixed with Bouin in the middle of the uterus, stained with H&E, and observed and photographed.

[0246] Because progesterone oil injection was administered daily, the final two testing time points were set at 1 hour before dosing on days 6 and 7. The results in Tables 24-25 show that despite daily injections, blood concentrations were only 1.922±0.642 ng / mL and 2.920±1.026 ng / mL in less than 24 hours. Composition 3, on the other hand, was administered only once, achieving blood concentrations of 2.688±1.291 ng / mL and 1.640±0.411 ng / mL at 144 and 168 hours, respectively, demonstrating a sustained-release effect.

[0247] Table 24 Concentration-time data of progesterone in plasma after subcutaneous injection of composition 3 (ng / mL) (7 days / time)

[0248] Table 25 Concentration-time data of progesterone in plasma after intramuscular injection of progesterone oil injection (ng / mL) (1 day / time)

[0249] *Data at 143h and 167h are data before the next injection; data at other times are data after administration.

[0250] Figure 3 shows that the progesterone oil injection group produced erythema at the injection site 7 days after administration, which was mildly irritating; while the progesterone pharmaceutical composition group and the solvent group showed no irritation, indicating that the irritation at the injection site was greatly reduced.

[0251] The results of the uterine weight gain experiment are shown in Figure 4(A) and Table 26. The uterine weight of the vehicle group was 0.228±0.066 g, and the uterine coefficient was 0.017±0.004. The uterine weight of the progesterone pharmaceutical composition group was 1.550±0.513 g, and the uterine coefficient was 0.116±0.039. The uterine weight of the progesterone oil injection group was 0.792±0.506 g, and the uterine coefficient was 0.059±0.039. Figure 4(B) shows that the progesterone concentration in the rabbit uterus in the vehicle group was 1.780±1.933 ng / g, and the progesterone concentration in the rabbit uterus in the progesterone pharmaceutical composition group was 107.54±60.922 ng / g, which was much higher than the progesterone concentration of 7.140±4.294 ng / g in the progesterone oil injection group. This strongly proves that the progesterone pharmaceutical composition group can maintain a higher concentration of progesterone in the uterus while greatly reducing the frequency of administration, thereby effectively exerting the role of progesterone in promoting endometrial development.

[0252] Table 26 Rabbit uterine weight and uterine coefficient

[0253] As shown in Figure 5, the red circled area shows the uterine glands. In the vehicle group, the number of glands was small, and only scattered glands under the endometrium were seen, and there was no obvious uterine gland structure in the deeper matrix. In the drug composition 3 group, the glands were heavily proliferated, the number increased, and the arrangement was crowded. The proliferated glands extended to the near muscle layer, and the common wall and back-to-back phenomena were seen. In the progesterone oil injection group, the glands were moderately proliferated, the number was large, and the uterine glands extended to the deeper matrix, and the common wall and back-to-back phenomena were seen.

[0254] In addition, as shown in Table 27, the number of uterine glands in the rabbits in the vehicle group was approximately 1.333 ± 1.269, the number of uterine glands in the Pharmaceutical Composition 3 group was approximately 30.933 ± 8.719, and the number of glands in the progesterone oil injection group was approximately 22.200 ± 13.146. The number of glands in the Pharmaceutical Composition 3 group was higher than that in the progesterone oil injection group, indicating that the progesterone pharmaceutical composition of the present invention is more effective in exerting its progestogen effect and promoting the increase of uterine glands.

[0255] Table 27 Number of uterine glands in rabbits

[0256] The above results further verify that the preparation has achieved the invention objectives of reducing injection frequency, reducing injection irritation and exerting excellent pharmacological effects proposed by the present invention.

[0257] Example 15

[0258] Moist heat sterilization

[0259] The composition 3 sample in Example 11 (the formulation composition and preparation process are detailed in Example 11) was sterilized by moist heat sterilization using a sterilization process of 121°C / 12 min. The properties and contents of the samples before and after sterilization are compared in the table below.

[0260] Table 28 Comparison of properties and contents of composition 3 before and after sterilization

[0261] The results showed no significant changes in the composition's properties and content before and after sterilization, demonstrating that the pharmaceutical composition of the present invention can be sterilized with moist heat at 121°C for 12 minutes. Compared to aseptic processing, moist heat sterilization offers advantages such as reliable sterilization, high efficiency, convenience, and ease of control, minimizing contamination risks.

[0262] The above description is merely an illustrative description of the present invention and its embodiments, which is not restrictive. The embodiment shown in the embodiment is only one embodiment of the present invention, and the actual embodiment is not limited thereto. Therefore, if a person skilled in the art is inspired by the above description and, without departing from the purpose of the present invention, designs an embodiment and examples similar to the technical solution without creatively designing, they shall fall within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition comprising the following components: a. An active ingredient; b. A sustained-release carrier; c. A lipophilic non-ionic surfactant; d. A pharmaceutically acceptable solvent; Among them, The active ingredient includes progesterone; The sustained-release carrier includes phospholipids and cholesterol; The pharmaceutically acceptable solvent includes a dispersion solvent; The dispersion solvent includes an organic solvent NMP and a short-chain alcohol; The active ingredient accounts for 5-13% (w / w) of the total weight of the pharmaceutical composition; The ratio of the phospholipids to the active ingredient is (3.5-10):1 (w / w); The ratio of the cholesterol to the active ingredient is (0.5-3.5):1 (w / w); The ratio of the dispersion solvent to the active ingredient is (2-8):1 (w / w); The ratio of the lipophilic non-ionic surfactant to the active ingredient is (0-1.2):1 (w / w); The pharmaceutically acceptable solvent optionally includes a phase transition regulator, and the weight percentage of the phase transition regulator in the composition is 0-4%.

2. The pharmaceutical composition according to claim 1, wherein the ratio of the cholesterol to the active ingredient is (1-2.5):1 (w / w), and more preferably the ratio of the cholesterol to the active ingredient is (1.5-2.5):1 (w / w).

3. The pharmaceutical composition according to any one of claims 1 or 2, wherein the ratio of the organic dispersion solvent to the active ingredient is (3-6):1 (w / w).

4. The pharmaceutical composition according to claim 1 or 2, wherein the short-chain alcohol includes one or more of ethanol, propylene glycol or glycerol.

5. The pharmaceutical composition according to claim 3, wherein the short-chain alcohol at least includes ethanol.

6. The pharmaceutical composition according to claim 5, wherein the ratio of NMP to ethanol in the organic dispersion solvent is (2-4):1 (w / w).

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the lipophilic non-ionic surfactant is selected from glyceryl monostearate (GMS).

8. The pharmaceutical composition according to claim 7, wherein the ratio of GMS to the active ingredient is (0-1):

1.

9. The pharmaceutical composition according to claim 1, comprising the following components: a. An active ingredient; b. A sustained-release carrier; c. A lipophilic non-ionic surfactant; d. A pharmaceutically acceptable solvent; Among them, The active ingredient includes progesterone; The sustained-release carrier includes phospholipids and cholesterol; The pharmaceutically acceptable solvent includes a dispersion solvent; The dispersion solvent includes an organic solvent NMP and a short-chain alcohol; The active ingredient accounts for 5-13% (w / w) of the total weight of the pharmaceutical composition; The ratio of the phospholipids to the active ingredient is (4-8):1 (w / w); The ratio of the cholesterol to the active ingredient is (1-2.5):1 (w / w); The ratio of the dispersion solvent to the active ingredient is (3-6):1 (w / w); The ratio of the lipophilic non-ionic surfactant to the active ingredient is (0-1):1 (w / w).

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the phase transition regulator includes water.

11. The pharmaceutical composition according to claim 10, wherein the water accounts for 0 - 4% of the total weight of the composition.

12. The pharmaceutical composition according to any one of claims 1 - 11, wherein the phospholipid is a natural phospholipid, including soybean phospholipid, lecithin or a combination thereof.

13. The pharmaceutical composition according to claim 12, wherein the content of phosphatidylcholine in the natural phospholipid is not less than 80%.

14. The pharmaceutical composition according to any one of claims 1 - 13, wherein the dosage form of the pharmaceutical composition is selected from injection solutions. Preferably, the injection solution includes subcutaneous injection or intramuscular injection.

15. The pharmaceutical composition according to any one of claims 1 - 14, wherein the concentration of progesterone in the pharmaceutical composition is: 55 - 130 mg / mL; preferably 60 - 100 mg / mL; more preferably 60 - 80 mg / mL.

16. An injection solution comprising the pharmaceutical composition according to any one of claims 1 - 15. Preferably, the injection solution is a subcutaneous injection solution or an intramuscular injection solution.

17. An in-situ gel comprising the pharmaceutical composition according to any one of claims 1 - 15.

18. A sustained-release depot drug comprising the pharmaceutical composition according to any one of claims 1 - 15.

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