Gemcitabine hydrochloride for injection and preparation method thereof

Abstract

The invention belongs to the technical field of antitumor drug preparations, and relates to gemcitabine hydrochloride for injection and a preparation method thereof. The gemcitabine hydrochloride for injection disclosed by the invention is prepared from gemcitabine hydrochloride, thiamine modified phosphatidylcholine, cholesterol and methyl glucoside sesquistearate. According to the invention, thiamine and phosphatidylcholine are combined as a carrier material, so that the stability of a liposome membrane is enhanced, the encapsulation efficiency is improved, more loading sites are provided for drugs, and the drug loading efficiency of the drugs is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of anti-tumor drug preparations, and specifically relates to gemcitabine hydrochloride for injection and a preparation method thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Cancer is one of the major diseases that seriously threaten human health in the world today, and its morbidity and mortality rates have remained high. Chemotherapy, as one of the important means of comprehensive cancer treatment, plays an indispensable role in the field of cancer treatment. Gemcitabine hydrochloride is a broad-spectrum antimetabolite anti-tumor drug with a chemical formula of C9H12ClF2N3O4. It is converted into active diphosphate and triphosphate nucleosides through the action of a series of enzymes in cells. These active metabolites can inhibit DNA synthesis, thereby preventing the proliferation and growth of tumor cells. Gemcitabine hydrochloride has shown significant therapeutic effects on a variety of solid tumors, such as non-small cell lung cancer, pancreatic cancer, breast cancer, etc., and occupies an important position in clinical tumor treatment.

[0004] However, gemcitabine hydrochloride faces many challenges during formulation development.

[0005] (1) Poor drug solubility: As a highly hydrophilic drug, gemcitabine has poor water solubility, which leads to low oral bioavailability and incomplete absorption, thus affecting the efficacy of the drug. In order to improve the efficacy of gemcitabine, it is usually necessary to prepare it into an injection form, but the existing preparation method of injection forms still faces certain technical bottlenecks.

[0006] (2) Poor drug stability: The chemical structure of gemcitabine hydrochloride is unstable and it is prone to degradation in aqueous solution, especially under high temperature, light, acidic or alkaline conditions, where its degradation rate will be significantly accelerated. This will not only lead to the loss of active ingredients of the drug and reduce the efficacy of the drug, but may also produce some unknown impurities and increase the risk of toxic side effects of the drug.

[0007] (3) Adverse reactions: Although gemcitabine has achieved remarkable results in anti-tumor treatment, it is often accompanied by some adverse reactions during its use, including bone marrow suppression, nausea, vomiting, liver and kidney function damage, etc. These adverse reactions limit the scope of use of gemcitabine, especially in weak patients, elderly patients or patients with other diseases, the adverse reactions of the drug may be more serious.

[0008] As an excellent drug carrier, liposomes have the advantages of good biocompatibility, increased drug therapeutic index, reduced toxicity, etc. It is expected that gemcitabine hydrochloride can be made into liposomes to overcome its existing defects and improve drug efficacy and safety. Chinese patent No. CN111437259A discloses a liposome composition containing gemcitabine or its salt, which is a liposome composition containing gemcitabine or its salt in liposomes, wherein the content of cholesterol relative to the total amount of lipids constituting the liposomes is 10 mol% to 35 mol%, and the osmotic pressure of the inner water phase of the liposome is 2 to 8 times the osmotic pressure of the outer water phase of the liposome. The Chinese patent with publication number CN117942305A discloses a gemcitabine liposome with high encapsulation rate, which includes the following components by weight: 0.1-0.7 parts of gemcitabine hydrochloride, 0.9-6 parts of hydrogenated bean phosphatidylcholine, 0.4-3 parts of cholesterol, 0.3-2 parts of phosphatidylethanolamine, 9-66 parts of ammonium sulfate, 1-8 parts of buffer citric acid / sodium citrate and 4-25 parts of acidity regulator. However, the drug loading of the gemcitabine liposome is only about 4%, which will limit the efficacy of single administration, prolong the treatment cycle and increase the cost of drug preparation. Summary of the invention

[0009] In order to overcome the shortcomings of the prior art, the present invention provides a stable gemcitabine hydrochloride for injection with high drug loading, high encapsulation rate, and the gemcitabine hydrochloride for injection comprises: 10 to 30 parts by weight of gemcitabine hydrochloride, 15 to 40 parts by weight of thiamine-modified phosphatidylcholine, 2 to 10 parts by weight of cholesterol, and 1 to 3 parts by weight of methyl glucoside sesquistearate.

[0010] The preparation method of the gemcitabine hydrochloride for injection is:

[0011] (1) The thiamine-modified phosphatidylcholine is ground and dissolved with cholesterol and methyl glucoside sesquistearate in a suitable amount of a mixed solvent of chloroform and ethanol in a volume ratio of 2 to 4:1, and then subjected to reduced pressure rotary evaporation at 35°C ± 5°C until a thin film is formed;

[0012] (2) dissolving gemcitabine hydrochloride in a buffer solution of pH = 4.5 to 6.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40° C. ± 5° C. for 20 to 40 min, performing high pressure homogenization, and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0013] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C ± 5°C, time 1 to 3 h, b. primary drying: temperature 10°C ± 5°C, pressure 10 to 100 Pa, time 12 to 20 h, c. secondary drying: temperature 15°C ± 5°C, pressure 1 to 10 Pa, time 6 to 12 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0014] Thiamine-modified phosphatidylcholine is prepared by solvent evaporation under acidic conditions from thiamine and phosphatidylcholine, specifically, by dropwise mixing of thiamine aqueous solution, phosphatidylcholine ethanol solution, and citric acid ethanol solution, standing, waiting for crystals to precipitate, suction filtration, washing, and drying. The molar ratio of thiamine, phosphatidylcholine, and citric acid is 2:2:1. It should be noted that the present invention has verified the combination of thiamine and phosphatidylcholine by infrared spectroscopy, and the infrared spectrum of thiamine-modified phosphatidylcholine has both characteristic peaks of thiamine and characteristic peaks of phosphatidylcholine, and is somewhat different from the infrared spectrum of a physical mixture of thiamine and phosphatidylcholine.

[0015] Thiamine contains multiple functional groups in its structure. Through interactions with phospholipids (such as hydrogen bonds, electrostatic effects, etc.), a large number of screening experiments have shown that thiamine is more suitable for binding to phosphatidylcholine, which can provide more loading sites for drugs and load drugs more effectively, thereby improving drug loading efficiency. It can also enhance the stability of the liposome membrane, increase the encapsulation rate of drugs, and allow more drugs to be encapsulated in the liposome.

[0016] Compared with the prior art, the technical effects of the present invention are:

[0017] The present invention optimizes the lipid carrier material, combines thiamine with phosphatidylcholine, can enhance the stability of the liposome membrane and improve the encapsulation rate in the preparation of gemcitabine hydrochloride liposomes, and also provides more loading sites for the drug, thereby improving the drug loading efficiency of the drug. In addition, the present invention also preferably selects a nonionic surfactant methyl glucoside sesquistearate, which can increase the steric hindrance between liposomes, prevent the liposomes from aggregating due to mutual collision, stabilize the liposome structure, and reduce the leakage rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Drug loading of gemcitabine hydrochloride for injection of Examples 1 to 3 and gemcitabine hydrochloride for injection of Comparative Examples 1 to 5.

[0019] Figure 2 : The encapsulation efficiency of gemcitabine hydrochloride for injection of Examples 1 to 3 and gemcitabine hydrochloride for injection of Comparative Examples 1 to 5.

[0020] Figure 3 : The leakage rates of gemcitabine hydrochloride for injection of Examples 1 to 3 and gemcitabine hydrochloride for injection of Comparative Examples 1 to 5. DETAILED DESCRIPTION

[0021] In order to make the purpose and technical solution of the present invention clearer, the present invention is further described below in conjunction with embodiments, but the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. It should be understood by those skilled in the art that any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0022] Preparation of Thiamine-modified Phosphatidylcholine

[0023] 10g thiamine is dissolved in 100ml water, 22.4g phosphatidylcholine is dissolved in 100ml ethanol, and 2.8g citric acid is dissolved in 50ml ethanol. Add the above three solutions dropwise and mix them. Let them stand at 22℃±5℃ for 10 days until crystals are precipitated. Then filter, wash and dry them to obtain the product.

[0024] Preparation of thiamine-modified phosphatidylethanolamine

[0025] 10g thiamine is dissolved in 100ml water, 21.8g phosphatidylethanolamine is dissolved in 100ml chloroform, and 2.8g citric acid is dissolved in 50ml ethanol. Add the three solutions dropwise and mix them. Let them stand at 22℃±5℃ for 10 days until crystals are precipitated. Then filter, wash and dry them to obtain the product.

[0026] Preparation of thiamine-modified phosphatidylserine

[0027] 10g thiamine is dissolved in 100ml water, 11.4g phosphatidylserine is dissolved in 100ml chloroform, 2.8g citric acid is dissolved in 50ml ethanol, the above three solutions are added dropwise and mixed, and the mixture is allowed to stand at 22℃±5℃ for 10 days until crystals are precipitated, which are then filtered, washed, and dried to obtain the product.

[0028] Example 1 Gemcitabine Hydrochloride for Injection

[0029] formula:

[0030] Dosage(g) Gemcitabine Hydrochloride 20 Thiamine-modified phosphatidylcholine 25 cholesterol 5 Methyl glucoside sesquistearate 2

[0031] Preparation method:

[0032] (1) The thiamine-modified phosphatidylcholine was ground and dissolved in 50 ml of a 3:1 chloroform-ethanol mixed solvent with cholesterol and methyl glucoside sesquistearate, and then evaporated under reduced pressure at 35°C ± 5°C until a thin film was formed;

[0033] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=5.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 30 min, performing high pressure homogenization (1100 psi, 2 min, 3 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0034] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C, time 2 h, b. primary drying: temperature 10°C, pressure 50 Pa, time 18 h, c. secondary drying: temperature 15°C, pressure 5 Pa, time 8 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0035] Example 2 Gemcitabine Hydrochloride for Injection

[0036] formula:

[0037] Dosage(g) Gemcitabine Hydrochloride 10 Thiamine-modified phosphatidylcholine 15 cholesterol 2 Methyl glucoside sesquistearate 1

[0038] Preparation method:

[0039] (1) The thiamine-modified phosphatidylcholine was ground and dissolved in 50 ml of a chloroform-ethanol mixed solvent with cholesterol and methyl glucoside sesquistearate in a volume ratio of 2:1, and then vacuum evaporated at 35°C ± 5°C until a thin film was formed;

[0040] (2) dissolving gemcitabine hydrochloride in a citrate buffer solution at pH = 4.5, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C ± 5°C for 20 min, performing high pressure homogenization (1000 psi, 1 min, 4 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0041] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -40°C, time 1 h, b. primary drying: temperature 5°C, pressure 100 Pa, time 12 h, c. secondary drying: temperature 10°C, pressure 10 Pa, time 6 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0042] Example 3 Gemcitabine Hydrochloride for Injection

[0043] formula:

[0044] Dosage(g) Gemcitabine Hydrochloride 30 Thiamine-modified phosphatidylcholine 40 cholesterol 10 Methyl glucoside sesquistearate 3

[0045] Preparation method:

[0046] (1) The thiamine-modified phosphatidylcholine was ground and dissolved in 50 ml of a chloroform-ethanol mixed solvent with cholesterol and methyl glucoside sesquistearate in a volume ratio of 4:1, and then vacuum evaporated at 35°C ± 5°C until a thin film was formed;

[0047] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=6.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 40 min, performing high pressure homogenization (1200 psi, 3 min, 2 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0048] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -30°C, time 3 h, b. primary drying: temperature 15°C, pressure 10 Pa, time 20 h, c. secondary drying: temperature 20°C, pressure 1 Pa, time 12 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0049] Comparative Example 1 Gemcitabine Hydrochloride for Injection

[0050] formula:

[0051] Dosage(g) Gemcitabine Hydrochloride 20 Thiamine-modified phosphatidylcholine 25 cholesterol 5

[0052] Preparation method:

[0053] (1) The thiamine-modified phosphatidylcholine was ground and dissolved with cholesterol in 50 ml of a 3:1 chloroform-ethanol mixed solvent, and then vacuum evaporated at 35°C ± 5°C until a thin film was formed;

[0054] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=5.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 30 min, performing high pressure homogenization (1100 psi, 2 min, 3 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0055] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C, time 2 h, b. primary drying: temperature 10°C, pressure 50 Pa, time 18 h, c. secondary drying: temperature 15°C, pressure 5 Pa, time 8 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0056] Comparative Example 2 Gemcitabine Hydrochloride for Injection

[0057] formula:

[0058]

[0059]

[0060] Preparation method:

[0061] (1) The thiamine-modified phosphatidylcholine was ground and dissolved in 50 ml of a 3:1 chloroform-ethanol mixed solvent with cholesterol and Tween-80, and then evaporated under reduced pressure at 35°C ± 5°C until a thin film was formed;

[0062] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=5.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 30 min, performing high pressure homogenization (1100 psi, 2 min, 3 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0063] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C, time 2 h, b. primary drying: temperature 10°C, pressure 50 Pa, time 18 h, c. secondary drying: temperature 15°C, pressure 5 Pa, time 8 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0064] Comparative Example 3 Gemcitabine Hydrochloride for Injection

[0065] formula:

[0066] Dosage(g) Gemcitabine Hydrochloride 20 Phosphatidylcholine 25 cholesterol 5 Methyl glucoside sesquistearate 2

[0067] Preparation method:

[0068] (1) Dissolve phosphatidylcholine, cholesterol, and methyl glucoside sesquistearate in 50 ml of a 3:1 chloroform-ethanol mixed solvent, and evaporate under reduced pressure at 35°C ± 5°C until a thin film is formed;

[0069] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=5.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 30 min, performing high pressure homogenization (1100 psi, 2 min, 3 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0070] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C, time 2 h, b. primary drying: temperature 10°C, pressure 50 Pa, time 18 h, c. secondary drying: temperature 15°C, pressure 5 Pa, time 8 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0071] Comparative Example 4 Gemcitabine Hydrochloride for Injection

[0072] formula:

[0073]

[0074]

[0075] Preparation method:

[0076] (1) The thiamine-modified phosphatidylethanolamine was ground and dissolved in 50 ml of a 3:1 chloroform-ethanol mixed solvent with cholesterol and methyl glucoside sesquistearate, and then evaporated under reduced pressure at 35°C ± 5°C until a thin film was formed;

[0077] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=5.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 30 min, performing high pressure homogenization (1100 psi, 2 min, 3 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0078] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C, time 2 h, b. primary drying: temperature 10°C, pressure 50 Pa, time 18 h, c. secondary drying: temperature 15°C, pressure 5 Pa, time 8 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0079] Comparative Example 5 Gemcitabine Hydrochloride for Injection

[0080] formula:

[0081] Dosage(g) Gemcitabine Hydrochloride 20 Thiamine-modified phosphatidylserine 25 cholesterol 5 Methyl glucoside sesquistearate 2

[0082] Preparation method:

[0083] (1) The thiamine-modified phosphatidylserine was crushed and ground, dissolved with cholesterol and methyl glucoside sesquistearate in 50 ml of a 3:1 chloroform-ethanol mixed solvent, and evaporated under reduced pressure at 35°C ± 5°C until a thin film was formed;

[0084] (2) dissolving gemcitabine hydrochloride in a phosphate buffer solution of pH=5.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40°C±5°C for 30 min, performing high pressure homogenization (1100 psi, 2 min, 3 cycles), and performing microfiltration to obtain gemcitabine hydrochloride liposomes;

[0085] (3) Freeze-drying the gemcitabine hydrochloride liposomes (a. pre-freezing: temperature -35°C, time 2 h, b. primary drying: temperature 10°C, pressure 50 Pa, time 18 h, c. secondary drying: temperature 15°C, pressure 5 Pa, time 8 h, d. sealing) to obtain gemcitabine hydrochloride for injection.

[0086] Quality evaluation of gemcitabine hydrochloride for injection of the present invention

[0087] Drug loading

[0088] Drug loading refers to the ratio of the weight of the drug in the liposome to the weight of the liposome (including the drug and lipid material), and the calculation formula is: drug loading (%) = (weight of the drug in the liposome / total weight of the liposome) × 100%. Drug loading is an important indicator for measuring the quality of drug-loaded liposomes and drug loading efficiency.

[0089] Figure 1The drug loading of the gemcitabine hydrochloride for injection of Examples 1 to 3 and the gemcitabine hydrochloride for injection of Comparative Examples 1 to 5. The gemcitabine hydrochloride for injection of Examples 1 to 3 of the present invention has a high drug loading, and a higher drug loading means that under the same dose of liposome administration, more drugs can be carried into the body, thereby improving the therapeutic effect of the drug.

[0090] Encapsulation efficiency

[0091] Encapsulation rate refers to the ratio of the amount of drug encapsulated in liposomes to the total amount of drug input, expressed as a percentage. It reflects the degree to which the drug is successfully encapsulated by the liposomes and is a key indicator for evaluating the quality of the preparation process of drug-loaded liposomes. The calculation formula is: Encapsulation rate (%) = (amount of drug in liposomes / total amount of drug input) × 100%.

[0092] Figure 2 The encapsulation rates of gemcitabine hydrochloride for injection of Examples 1 to 3 and gemcitabine hydrochloride for injection of Comparative Examples 1 to 5 are shown. The gemcitabine hydrochloride for injection of Examples 1 to 3 of the present invention has a high encapsulation rate, which indicates that the preparation method can effectively encapsulate the drug in the liposome, reduce the loss of the drug during the preparation process, and is conducive to ensuring the stability of the drug during storage and transportation, and also has an important influence on the release and bioavailability of the drug.

[0093] Leakage rate

[0094] The leakage rate refers to the ratio of the amount of drug leaked from the liposome under certain conditions (such as during storage) to the amount of drug initially encapsulated in the liposome. It reflects the ability of the liposome to retain the drug and is an important indicator for measuring the stability of the drug-loaded liposome. Calculation formula: leakage rate (%) = (amount of drug leaked / amount of drug initially encapsulated in the liposome) × 100%.

[0095] The gemcitabine hydrochloride for injection of Examples 1 to 3 and the gemcitabine hydrochloride for injection of Comparative Examples 1 to 5 were placed in an environment with a temperature of 40° C.±2° C. and a relative humidity of 75%±5% for 90 days, and the leakage rate was evaluated. Figure 3 The leakage rates of gemcitabine hydrochloride for injection of Examples 1 to 3 and gemcitabine hydrochloride for injection of Comparative Examples 1 to 5 are shown. The leakage rates of gemcitabine hydrochloride for injection of Examples 1 to 3 of the present invention are low. The low leakage rate indicates that the liposome can effectively keep the drug inside, and the drug is not easy to leak during storage and transportation, thereby ensuring the effectiveness and safety of the drug. The leakage rate of gemcitabine hydrochloride for injection of Comparative Examples 1 to 5 is too high, which will cause a large amount of drug to leak before reaching the target site, thereby reducing the therapeutic effect of the drug.

Claims

1. A gemcitabine hydrochloride for injection, characterized in that: The gemcitabine hydrochloride for injection comprises: 10 to 30 parts by weight of gemcitabine hydrochloride, 15 to 40 parts by weight of thiamine-modified phosphatidylcholine, 2 to 10 parts by weight of cholesterol, and 1 to 3 parts by weight of methyl glucoside sesquistearate.

2. The gemcitabine hydrochloride for injection according to claim 1, characterized in that: The thiamine-modified phosphatidylcholine is prepared by using thiamine and phosphatidylcholine through a solvent evaporation method under acidic conditions.

3. The gemcitabine hydrochloride for injection according to claim 1, characterized in that: The preparation method of the thiamine-modified phosphatidylcholine is as follows: dropwise adding a thiamine aqueous solution, a phosphatidylcholine ethanol solution and a citric acid ethanol solution, mixing them, standing them, waiting for crystals to precipitate, filtering them by suction, washing them and drying them.

4. The gemcitabine hydrochloride for injection according to claim 1, characterized in that: The molar ratio of thiamine, phosphatidylcholine and citric acid is 2:2:

1.

5. The gemcitabine hydrochloride for injection according to claim 1, characterized in that: The gemcitabine hydrochloride for injection comprises: 20 parts by weight of gemcitabine hydrochloride, 25 parts by weight of thiamine-modified phosphatidylcholine, 5 parts by weight of cholesterol, and 2 parts by weight of methyl glucoside sesquistearate.

6. A method for preparing gemcitabine hydrochloride for injection according to claim 1, characterized in that: The preparation method is: (1) grind thiamine-modified phosphatidylcholine, dissolve it in an appropriate amount of mixed solvent with cholesterol and methyl glucoside sesquistearate, and evaporate it under reduced pressure at 35°C ± 5°C until a thin film is formed; (2) dissolving gemcitabine hydrochloride in a buffer solution of pH = 4.5 to 6.0, adding the solution to the film of step (1) for hydration, performing vacuum rotary evaporation at 40° C. ± 5° C. for 20 to 40 min, performing high pressure homogenization, and performing microfiltration to obtain gemcitabine hydrochloride liposomes; (3) freeze-drying the gemcitabine hydrochloride liposome to obtain gemcitabine hydrochloride for injection.

7. The method for preparing gemcitabine hydrochloride for injection according to claim 6, characterized in that: The mixed solvent is a mixed solvent of chloroform and ethanol in a volume ratio of 2 to 4:

1.

8. The method for preparing gemcitabine hydrochloride for injection according to claim 6, characterized in that: The buffer solution is one of a phosphate buffer solution and a citrate buffer solution.

9. The method for preparing gemcitabine hydrochloride for injection according to claim 6, characterized in that: The pH=5.

0.

10. The method for preparing gemcitabine hydrochloride for injection according to claim 6, characterized in that: The freeze drying procedure is: a. Pre-freezing: temperature -35℃±5℃, time 1~3h, b. Primary drying: temperature 10℃±5℃, pressure 10~100Pa, time 12~20h, c. Secondary drying: temperature 15℃±5℃, pressure 1~10Pa, time 6~12h, d. Sealing.

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