Preparation method of anti-hepatic fibrosis medicine containing N-acetylcitrulline

Abstract

The invention discloses a preparation method of an anti-hepatic fibrosis drug containing N-acetylcitrulline, and relates to the technical field of pharmaceutical preparations, and the preparation method comprises the following steps: S1, providing N-acetylcitrulline as an active component; s2, mixing the N-acetylcitrulline with one or more pharmaceutically acceptable auxiliary materials, so as to form a uniform mixture; and S3, processing the mixture into a pharmaceutical preparation suitable for administration. According to the invention, the medication compliance and the process amplification are improved, or the mixture is prepared into an injection which comprises the steps of dissolution, pH regulation, sterilization and filtration and sterilization, so that the sterility and the physical and chemical stability of the medicine are ensured, and the injection is suitable for intravenous administration of acute and critical patients, thereby expanding clinical application scenarios on the aspect of dosage forms, and improving the clinical application prospect. And finally, the beneficial effects that the new application is found and converted into a practical medicine product which can be industrially produced and has good safety and definite anti-fibrosis effect are achieved.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, and in particular to a method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline. Background Technology

[0002] Liver fibrosis is the core pathological stage in the progression of chronic liver disease to cirrhosis and hepatocellular carcinoma, and it constitutes a major factor in liver disease-related deaths worldwide. Current clinical intervention strategies mainly focus on etiological control, such as antiviral therapy and alcohol abstinence, which have limited efficacy in reversing existing fibrotic tissue. In the field of direct antifibrotic drug development, inhibitors of key signaling pathways such as transforming growth factor-β and platelet-derived growth factor, or collagen metabolism regulators, have become the direction of exploration. However, most candidate drugs are still in the preclinical or early clinical research stage, and generally face challenges such as uncertain efficacy, insufficient target specificity, or potential systemic toxic side effects. Although mesenchymal stem cell-based cell therapy can theoretically regulate hepatic stellate cell function through paracrine effects, it suffers from technical bottlenecks such as low in vivo colonization rate, short survival time, complex preparation process, and unstable efficacy, which seriously restricts its clinical translation prospects. Developing a drug with a novel mechanism of action that can effectively intervene in the fibrosis process from the source and has good safety has become a key technical problem that urgently needs to be solved in this field.

[0003] In the existing technology, the exploration of endogenous substances and their derivatives in the field of anti-liver fibrosis is not yet in-depth. Taking N-acetylcitrulline as an example, its known technical uses mainly revolve around its role as a nitric oxide precursor in improving vascular endothelial function or as a sports nutrition supplement. Based on its established metabolic pathway, the existing technology has not revealed that this compound has a direct inhibitory effect on the activation and proliferation of hepatic stellate cells, the core effector cells of liver fibrosis. This gap in knowledge has led to a lack of clear technical inspiration in the field for developing N-acetylcitrulline into an anti-liver fibrosis drug. The existing technology has failed to provide any teaching that this compound can intervene in the key initiation of the fibrosis process of hepatic stellate cell activation, thus making it impossible to have a feasible technical basis for constructing a highly effective and low-toxicity anti-fibrosis drug based on this compound. Summary of the Invention

[0004] In view of the aforementioned existing problems, the present invention is proposed.

[0005] Therefore, the present invention provides a method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline, which solves the technical problem that the prior art could not develop anti-hepatic fibrosis drugs based on compounds because it failed to reveal the new use of N-acetylcitrulline in inhibiting the activation of hepatic stellate cells.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution: In a first aspect, the present invention provides a method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline, comprising: S1, providing N-acetylcitrulline as an active ingredient; S2. The N-acetylcitrulline is mixed with one or more pharmaceutically acceptable excipients to form a homogeneous mixture; S3. Process the mixture into a pharmaceutical preparation suitable for administration.

[0007] In a preferred embodiment of the preparation method of the anti-hepatic fibrosis drug containing N-acetylcitrulline according to the present invention, the drug preparation is an oral preparation.

[0008] In a preferred embodiment of the preparation method of the N-acetylcitrulline-containing anti-hepatic fibrosis drug of the present invention, the oral formulation is a tablet comprising: S2a. Mix the N-acetylcitrulline with the filler and disintegrant until homogeneous; S2b: Add a binder solution to the mixed material, form a soft mass, granulate, and dry to obtain dry granules; S2c. Add lubricant to the dried particles and mix thoroughly. S3a. Compress the mixed granules into tablets.

[0009] In a preferred embodiment of the preparation method of the anti-hepatic fibrosis drug containing N-acetylcitrulline according to the present invention, the filler is selected from one or more of microcrystalline cellulose, lactose, pregelatinized starch, and mannitol. The disintegrant is selected from one or more of croscarmellose sodium, croscarmellose, and carboxymethyl starch sodium.

[0010] As an excellent method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in this invention Option 1: The adhesive is selected from one or more of hydroxypropyl methylcellulose, povidone, and starch paste. The lubricant is selected from one or more of magnesium stearate, sodium stearate fumarate, and micronized silica gel.

[0011] In a preferred embodiment of the preparation method of the anti-hepatic fibrosis drug containing N-acetylcitrulline according to the present invention, in step S2b, the drying temperature is 60°C to 70°C, and the moisture content of the dried particles is controlled at 2.0% to 3.0%.

[0012] In a preferred embodiment of the preparation method of the anti-hepatic fibrosis drug containing N-acetylcitrulline according to the present invention, the drug preparation is an injection.

[0013] In a preferred embodiment of the preparation method of the N-acetylcitrulline-containing anti-hepatic fibrosis drug of the present invention, steps S2 and S3 include: S2d. Dissolve the N-acetylcitrulline in a portion of water for injection and stir until completely dissolved. S2e, adjust the pH of the solution to 6.0-7.5, and add water for injection to the full volume; S3b. The liquid medicine is sterilized, filtered, filled, sealed, and sterilized to obtain an injectable solution.

[0014] In a preferred embodiment of the preparation method of the N-acetylcitrulline-containing anti-hepatic fibrosis drug of the present invention, in step S3b, the sterilization filtration uses a 0.22 μm microporous membrane; the sterilization is performed using a moist heat sterilization method at 115°C for 30 minutes. As a preferred embodiment of the preparation method of the anti-hepatic fibrosis drug containing N-acetylcitrulline according to the present invention, the concentration of N-acetylcitrulline in the final drug formulation is from 1 mg / mL to 500 mg / mL, or accounts for 1% to 90% of the total weight of the formulation.

[0015] The beneficial effects of this invention are as follows: By providing the active ingredient N-acetylcitrulline, and based on its newly discovered medical use of inhibiting hepatic stellate cell activation, it is mixed with pharmaceutically acceptable excipients to form a homogeneous mixture, and then processed into a drug formulation suitable for administration. This innovatively transforms a known compound into an anti-hepatic fibrosis drug with clear targeting and source intervention effects. Furthermore, by further preparing the mixture into oral tablets, including the steps of mixing the active ingredient with specific fillers and disintegrants, granulation, drying, and adding lubricants before tableting, uniform distribution of the drug components, stable shaping, and convenient long-term oral administration to patients are achieved, improving medication adherence and process scalability. Alternatively, by preparing the mixture into an injectable form, including dissolution, pH adjustment, sterile filtration, and sterilization, the sterility and physicochemical stability of the drug are ensured, making it suitable for intravenous administration to critically ill patients. This expands the clinical application scenarios at the dosage form level, ultimately achieving the beneficial effect of transforming a new use discovery into a practical drug product that can be industrially produced, has good safety, and clear anti-fibrotic efficacy. Attached Figure Description

[0016] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the following description of the embodiments will be briefly introduced. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1This is a flowchart illustrating the preparation method of an anti-hepatic fibrosis drug containing N-acetylcitrulline. Detailed Implementation

[0018] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0019] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0020] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that is mutually exclusive with other embodiments.

[0021] Example 1, referring to Figure 1 This is the first embodiment of the present invention, which provides a method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline, comprising the following steps: Prescription composition (per 1000 tablets) N-acetylated citrulline: 50.0 g Microcrystalline cellulose (filler): 70.0 g Croscarmellose sodium (disintegrant): 10.0 g Hydroxypropyl methylcellulose (5% aqueous solution, binder): Appropriate amount Magnesium stearate (lubricant): 1.5 g Preparation process S1 (Provide active ingredient): Take 50.0g of N-acetylcitrulline raw material that meets pharmaceutical standards, and set aside.

[0022] S2a (mixing): N-acetylcitrulline, microcrystalline cellulose, and cross-linked sodium carboxymethyl cellulose are passed through an 80-mesh sieve and then placed in a three-dimensional motion mixer and mixed for 20 minutes to obtain a uniform powder mixture.

[0023] S2b (Granulation and Drying): Slowly add an appropriate amount of 5% hydroxypropyl methylcellulose aqueous solution as a binder to the above powder mixture while stirring to obtain a soft material with suitable hardness. Granulate the soft material through a 14-mesh sieve, and place the resulting wet granules in a hot air circulating oven to dry at 65°C until the moisture content of the granules drops to 2.5%.

[0024] S2c (total mixing): The dried granules are sized by passing them through a 16-mesh sieve, and then the sieved magnesium stearate is added. The mixture is then mixed in a mixer for 5 minutes to ensure thorough mixing.

[0025] S3a (Compression): Place the mixed granules in a rotary tablet press, adjust the pressure, and compress them into round tablets with a weight of approximately 131.5 mg.

[0026] Example 2, refer to Figure 1 This is a second embodiment of the present invention, which provides a method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline, comprising the following steps: Preparation of N-acetylicitrulline injection 1. Formula composition (based on 10 mL per vial): N-acetylcitrulline: 2.0 g, sodium hydroxide / hydrochloric acid solution (pH adjuster): appropriate amount, water for injection: add to 10 mL.

[0027] Preparation process S1 (Provide active ingredient): Take 2.0g of N-acetylcitrulline raw material that meets the standards for injection, and set aside.

[0028] S2d (Resolution Preparation): In the solution preparation tank, add approximately 80% of the total volume of water for injection, and slowly add N-acetylcitrulline while stirring until completely dissolved.

[0029] S2e (pH Adjustment and Volume Adjustment): Monitor the pH value of the solution with a pH meter, add 0.1 mol / L sodium hydroxide solution dropwise, and precisely adjust the pH to 6.8. Add water for injection to the total volume and stir well.

[0030] S3b (Sterilization and Filling): The above-mentioned drug solution is sterilized by sequentially passing it through 0.45μm and 0.22μm microporous membranes. Under aseptic conditions, the filtrate is filled into 10mL sterile ampoules, each containing 10.2mL, and immediately sealed.

[0031] Sterilization: Place the filled ampoules in a high-pressure steam sterilizer and sterilize at 115°C for 30 minutes.

[0032] Inspection and Packaging: After sterilization, leak detection and visible foreign matter inspection are carried out, and qualified products are labeled and packaged.

[0033] Quality Inspection The resulting injection solution was a colorless and clear liquid. Testing showed a pH of 6.8, sterility, pyrogen-free properties, and related substance content meeting regulations. The N-acetylic acid content was 101.2% of the labeled amount.

[0034] Example 3, referring to Figure 1This is the third embodiment of the present invention, which provides a method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline, comprising the following steps: Preparation of high-dose N-acetylcitrulline capsules Formula composition (per 1000 capsules): N-acetylicitrulline: 100.0 g, pregelatinized starch (filler): 50.0 g, sodium stearate fumarate (lubricant): 1.0 g.

[0035] Preparation process S1: Take 100.0g of N-acetylcitrulline.

[0036] S2: Sift N-acetylcitrulline and pregelatinized starch and mix them evenly. Then add sodium stearate fumarate and mix for 5 minutes.

[0037] S3: Fill the well-mixed powder into hollow capsules of appropriate size to obtain capsules.

[0038] Quality Inspection The contents of the capsules were mixed evenly, and the difference in filling volume met the regulations.

[0039] Comparative Example 1: Blank tablets without active ingredients Blank tablets were prepared according to the formulation and process of Example 1, but without the addition of N-acetylcitrulline as the active ingredient, using only microcrystalline cellulose to make up the weight. This comparative example is used to demonstrate that the drug efficacy derives from N-acetylcitrulline, rather than the excipients themselves.

[0040] Comparative Example 2: Tablets with citrulline as the active ingredient Tablets were prepared according to the formulation and process of Example 1, but with an equal amount of N-acetylcitrulline replaced by an equal molar amount of citrulline. This comparative example is intended to demonstrate that N-acetylcitrulline may have unique and unexpected superior effects (such as higher stability, bioavailability, or specificity) in anti-liver fibrosis compared to its parent compound citrulline.

[0041] Comparative Example 3: Tablets with Changed Key Process Parameters According to the formulation of Example 1, but during the drying process in step S2b, the drying temperature was increased to 85°C, resulting in a particle moisture content of less than 1.0%. Although the resulting tablets had high hardness, the disintegration time was significantly prolonged, affecting drug dissolution and potential bioavailability. This comparative example is used to highlight the crucial role of controlling specific process parameters of drying temperature and moisture (e.g., 60-70°C, moisture 2.0%-3.0%) in ensuring the quality of the finished product.

[0042] In summary, this invention provides the active ingredient N-acetylcitrulline, and based on its newly discovered medical use of inhibiting hepatic stellate cell activation, mixes it with pharmaceutically acceptable excipients to form a homogeneous mixture, and then processes it into a drug formulation suitable for administration. This innovatively transforms a known compound into an anti-hepatic fibrosis drug with clear targeting and source intervention effects. Specifically, by further preparing the mixture into oral tablets, including mixing the active ingredient with specific fillers and disintegrants, granulation, drying, and adding lubricants before tableting, uniform distribution of the drug components, stable shaping, and convenient long-term oral administration are achieved, improving medication adherence and process scalability. Alternatively, by preparing the mixture into an injectable form, including dissolution, pH adjustment, sterile filtration, and sterilization, the sterility and physicochemical stability of the drug are ensured, making it suitable for intravenous administration to critically ill patients. This expands the clinical application scenarios at the dosage form level, ultimately achieving the beneficial effect of transforming a new use discovery into a practical drug product that can be industrially produced, possesses good safety, and has clear anti-fibrotic efficacy.

[0043] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A method for preparing an anti-hepatic fibrosis drug containing N-acetylcitrulline, characterized in that: include, S1 provides N-acetylcitrulline as an active ingredient; S2. The N-acetylcitrulline is mixed with one or more pharmaceutically acceptable excipients to form a homogeneous mixture; S3. Process the mixture into a pharmaceutical preparation suitable for administration.

2. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 1, characterized in that: The pharmaceutical preparation is an oral preparation.

3. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 2, characterized in that: The oral preparation is a tablet, comprising: S2a. Mix the N-acetylcitrulline with the filler and disintegrant until homogeneous; S2b: Add a binder solution to the mixed material, form a soft mass, granulate, and dry to obtain dry granules; S2c. Add lubricant to the dried particles and mix thoroughly. S3a. Compress the mixed granules into tablets.

4. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 3, characterized in that: The filler is selected from one or more of microcrystalline cellulose, lactose, pregelatinized starch, and mannitol; The disintegrant is selected from one or more of croscarmellose sodium, croscarmellose, and carboxymethyl starch sodium.

5. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 3, characterized in that: The adhesive is selected from one or more of hydroxypropyl methylcellulose, povidone, and starch paste; The lubricant is selected from one or more of magnesium stearate, sodium stearate fumarate, and micronized silica gel.

6. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 3, characterized in that: In step S2b, the drying temperature is 60°C to 70°C, and the moisture content of the dried particles is controlled at 2.0% to 3.0%.

7. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 1, characterized in that: The pharmaceutical preparation is an injectable formulation.

8. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 7, characterized in that, Steps S2 and S3 include: S2d. Dissolve the N-acetylcitrulline in a portion of water for injection and stir until completely dissolved. S2e, adjust the pH of the solution to 6.0-7.5, and add water for injection to the full volume; S3b. The liquid medicine is sterilized, filtered, filled, sealed, and sterilized to obtain an injectable solution.

9. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 8, characterized in that: In step S3b, the sterilization filtration uses a 0.22μm microporous membrane; the sterilization uses a moist heat sterilization method at 115℃ for 30 minutes.

10. The method for preparing the anti-hepatic fibrosis drug containing N-acetylcitrulline as described in claim 9, characterized in that, The N-acetylcitrulline is present in a concentration of 1 mg / mL to 500 mg / mL in the final pharmaceutical formulation, or accounts for 1% to 90% of the total weight of the formulation.

Images