Rolapitant fat emulsion injection as well as preparation method and application thereof

Abstract

The invention provides a rolapitant fat emulsion injection as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The rolapitant fat emulsion injection provided by the invention comprises an oil phase component and a water phase component, the oil phase component comprises 0.1-1% of rolapitant, 0.1-5% of a cosolvent, 5-10% of vegetable oil and 1-2% of an emulsifier; the water phase component comprises 1-2.25% of an osmotic pressure regulator, 0.01-1% of an auxiliary emulsifier and the balance of water; the water is used, so that the total mass percentage of the oil phase component and the water phase component is 100%. According to the rolapitant fat emulsion injection disclosed by the invention, the use of high-concentration HS-15 with a hemolysis risk is avoided, and auxiliary materials with good safety are adopted, so that the potential toxic and side effect risk is reduced, the vomiting of a patient can be quickly relieved, and the clinical medication risk is remarkably reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a rolapitant fat emulsion injection, a preparation method thereof, and an application thereof. Background Art

[0002] Chemotherapy-induced nausea and vomiting (CINV) severely impacts patients' quality of life and treatment compliance. Internationally, chemotherapy drugs are classified into four categories based on their emetic risk: high (>90%), moderate (30%-90%), low (10%-30%), and mild (<10%). CINV is primarily categorized by onset time: acute (≤24 hours after treatment) and delayed (≥24 hours to 120-168 hours after treatment). 5-hydroxytryptamine (5-HT) is a key neurotransmitter in the pathogenesis of acute CINV; substance P (SP) and neurokinin-1 are the neurotransmitters most closely associated with delayed CINV. Clinically recommended methods for preventing and treating CINV caused by chemotherapy drugs with a high risk of emesis mainly include the combined use of NK-1 receptor antagonists, 5-HT3 receptor antagonists, and dexamethasone. For patients receiving chemotherapy drugs with a moderate risk of emesis, the combination of 5-HT3 receptor antagonists and dexamethasone with or without NK-1 receptor antagonists is generally recommended.

[0003] The neurotransmitter SP is a neuropeptide composed of 11 amino acids and is widely distributed in the central and peripheral nervous systems, as well as immune cells outside the nervous system, the liver, lungs, and the placenta. SP is an endogenous ligand for the NK-1 receptor, mediating vomiting, emotional behavior, depression, epilepsy, and neurodegenerative diseases through the NK-1 receptor. It is also associated with various inflammatory immune responses such as pain and asthma. The NK-1 receptor is a G protein-coupled receptor found in peripheral tissues, vascular endothelial cells, muscle cells, gastrointestinal tract, and urinary tract. Currently marketed NK-1 receptor antagonists such as aprepitant and its prodrug fosaprepitant can effectively prevent nausea and vomiting caused by various emetics, and also have a good preventive effect on nausea and vomiting caused by surgery, but there is a potential risk of interaction with CYP3A4.

[0004] Rolapitant, developed by Schering-Plough, is a long-acting, highly selective NK-1 receptor antagonist with a half-life of up to 180 hours. It antagonizes intracellular calcium release induced by NK-1 receptor agonists and inhibits G protein-coupled receptor signaling. OPKO Health is conducting post-Phase II clinical studies for this drug, and Tesaro is in charge of its marketing application. On September 1, 2015, rolapitant received marketing approval from the US Food and Drug Administration (FDA) in a 90 mg tablet under the trade name Varubi. It is indicated in combination with other antiemetics for the prevention of delayed nausea and vomiting associated with the initial or subsequent course of anti-cancer chemotherapy (including but not limited to chemotherapy with a high risk of emetogenicity). On October 25, 2017, rolapitant hydrochloride intravenous emulsion (Varubi) was approved for marketing in the US in a 166.5 mg / 92.5 mL dosage form. Clinical data indicate that the development of an intravenous emulsion of rolapitant has a shorter time to peak efficacy, faster onset of action, and enhanced efficacy and reduced toxicity compared to its oral formulation. Furthermore, the intravenous emulsion of rolapitant allows for a lower dosage and extends its half-life to seven days. However, rolapitant hydrochloride emulsion is prone to adverse reactions such as allergic reactions and anaphylactic shock within minutes of injection. Some patients experience symptoms such as difficulty breathing, facial or throat swelling, and hives, and even hypotension and anaphylactic shock, which can be life-threatening. Consequently, the FDA announced the withdrawal of rolapitant hydrochloride intravenous emulsion from the market in 2018.

[0005] Rolapitant hydrochloride intravenous emulsion contains 0.2% rolapitant hydrochloride monohydrate, 1.1% medium-chain triglycerides (MCT), 0.65% soybean oil, 4.4% polyethylene glycol-15 hydroxystearic acid (HS-15), 0.28% anhydrous sodium dihydrogen phosphate, 0.62% sodium chloride, water for injection, and hydrochloric acid or sodium hydroxide to adjust the pH to 7.0-8.0. HS-15 is a nonionic solubilizer and hemolytic. Excessive use can cause serious toxic side effects. HS-15 also contains nearly 30% free polyethylene glycol (PEG), which can increase histamine levels in the body and may be the cause of allergies. Summary of the Invention

[0006] The object of the present invention is to provide a rolapitant fat emulsion injection and a preparation method and application thereof, wherein the rolapitant fat emulsion injection is safe, has a low risk of toxic and side effects, and has good stability.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a rolapitant fat emulsion injection, comprising an oil phase component and an aqueous phase component;

[0009] The oil phase components include: 0.1-1% of rolapitant, 0.1-5% of cosolvent, 5-10% of vegetable oil, and 1-2% of emulsifier;

[0010] The aqueous phase components include: 1-2.25% osmotic pressure regulator, 0.01-1% auxiliary emulsifier, and the balance water;

[0011] The amount of water used is such that the total mass percentage of the oil phase component and the water phase component is 100%.

[0012] Preferably, the cosolvent includes one or more of ethanol, polyethylene glycol, glycerol and ethylene glycol.

[0013] Preferably, the vegetable oil comprises one or more of soybean oil, medium-chain oil, olive oil, castor oil, sesame oil and hydrogenated vegetable oil.

[0014] Preferably, the emulsifier includes one or more of egg yolk lecithin, egg yolk phosphatidylcholine, soybean lecithin, polyene phosphatidylcholine and hydrogenated phospholipids.

[0015] Preferably, the osmotic pressure regulator includes one or more of glycerol, sodium chloride, glucose, phosphate and citrate.

[0016] Preferably, the auxiliary emulsifier includes one or more of oleic acid, sodium oleate and phosphatidylglycerol.

[0017] The present invention provides a method for preparing the rolapitant fat emulsion injection described in the above technical solution, comprising the following steps:

[0018] mixing rolapitant and a cosolvent, and mixing the resulting mixture with an emulsifier and vegetable oil to obtain an oil phase;

[0019] mixing water, an osmotic pressure regulator, and a co-emulsifier to obtain an aqueous phase;

[0020] The aqueous phase and the oil phase are combined under shear conditions to obtain colostrum;

[0021] The colostrum is sequentially subjected to high-pressure homogenization and sterilization to obtain rolapitant fat emulsion injection.

[0022] Preferably, the pressure of the high-pressure homogenization is 10,000 to 20,000 psi.

[0023] The present invention provides the use of the rolapitant fat emulsion injection described in the above technical solution or the rolapitant fat emulsion injection prepared by the preparation method described in the above technical solution in the preparation of anti-nausea and vomiting drugs.

[0024] The present invention provides a rolapitant fat emulsion injection, wherein a cosolvent can increase the solubility of rolapitant, a vegetable oil is used as the oil phase of the emulsion, an emulsifier has good surface activity during the emulsification process, can reduce the surface tension between liquids, and form emulsion droplets, an osmotic pressure regulator can adjust the osmotic pressure, and an auxiliary emulsifier makes the surface of the emulsion droplets negatively charged. Due to the repulsive effect of like charges, the emulsion droplets are not easy to aggregate, thereby increasing the stability of the emulsion, stabilizing the surface Zeta potential value between -25 and -40 mV, and the injection has good stability and is equivalent to the original rolapitant hydrochloride intravenous emulsion (Varubi). The rolapitant fat emulsion injection of the present invention avoids the use of a higher concentration of HS-15 with a hemolytic risk, adopts an excipient with good safety, reduces the risk of potential toxic and side effects, can quickly relieve vomiting in patients, and significantly reduces the risk of clinical medication.

[0025] The rolapitant fat emulsion injection of the present invention has a milky white or translucent appearance, an average particle size of less than 200 nm, stable quality, and reduces adverse reactions such as hemolysis caused by free drugs and excipients.

[0026] The rolapitant fat emulsion injection of the present invention is prepared by an emulsification and homogenization method, and the preparation process is mature, reproducible, and stable, and can be produced in large quantities. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The drug concentration-time curves of Rolapitant injection prepared in Example 1 and Comparative Example 1 are shown;

[0028] Figure 2 This is a semi-logarithmic curve graph of the rolapitant injection prepared in Example 1 and Comparative Example 1;

[0029] Figure 3 This is a diagram of the hemolytic test process of the rolapitant injection prepared in Example 1 (before incubation, incubation for 15 minutes, 30 minutes, and 45 minutes);

[0030] Figure 4 This is a diagram of the hemolytic test process of the rolapitant injection prepared in Example 1 (incubation for 1 h, 2 h, 3 h, and 8 min after the end of incubation);

[0031] Figure 5 This is a diagram of the hemolytic test process of the rolapitant injection prepared in Comparative Example 1 (before incubation, incubation for 15 minutes, 30 minutes, and 45 minutes);

[0032] Figure 6 This is a diagram of the hemolytic test process of the rolapitant injection prepared in Comparative Example 1 (incubation for 1 hour, 2 hours, 3 hours, and 7 minutes after the end of incubation). DETAILED DESCRIPTION

[0033] The present invention provides a rolapitant fat emulsion injection, comprising an oil phase component and an aqueous phase component;

[0034] The oil phase components include: 0.1-1% of rolapitant, 0.1-5% of cosolvent, 5-10% of vegetable oil, and 1-2% of emulsifier;

[0035] The aqueous phase components include: 1% to 2.25% of an osmotic pressure regulator, 0.01% to 1% of an auxiliary emulsifier, and the balance of water;

[0036] The amount of water used is such that the total mass percentage of the oil phase component and the water phase component is 100%.

[0037] In the present invention, unless otherwise specified, the raw materials required for preparation are commercially available products well known to those skilled in the art.

[0038] The rolapitant fat emulsion injection provided by the present invention comprises an oil phase component; the oil phase component comprises: 0.1-1% of rolapitant, 0.1-5% of a cosolvent, 5-10% of a vegetable oil, and 1-2% of an emulsifier.

[0039] In the present invention, the mass percentage of rolapitant is 0.1-1%, preferably 0.2-0.8%, more preferably 0.3-0.6%; the rolapitant is preferably used in the form of rolapitant hydrochloride monohydrate.

[0040] In the present invention, the mass percentage of the cosolvent is 0.1-5%, preferably 0.5-2.5%. The cosolvent preferably includes one or more of ethanol, polyethylene glycol, glycerol, and ethylene glycol. When the cosolvent is two or more of the above cosolvents, the present invention has no particular limitation on the ratio of different types of cosolvents, and can be adjusted according to actual needs.

[0041] In the present invention, the mass percentage of the vegetable oil is 5-10%, preferably 10%. The vegetable oil preferably includes one or more of soybean oil, medium-chain oil, olive oil, castor oil, sesame oil, and hydrogenated vegetable oil; the medium-chain oil is preferably a medium-chain triglyceride. When the vegetable oil is two or more of the above, the present invention does not specifically limit the ratio of the different types of vegetable oils and can be adjusted according to actual needs.

[0042] In the present invention, the mass percentage of the emulsifier is 1-2%, preferably 1.2-1.5%. The emulsifier preferably includes one or more of egg yolk lecithin, egg yolk phosphatidylcholine, soybean lecithin, polyene phosphatidylcholine, and hydrogenated phospholipids. When the emulsifier is two or more of the above, the present invention has no particular limitation on the ratio of different types of emulsifiers, and can be adjusted according to actual needs.

[0043] The rolapitant fat emulsion injection provided by the present invention comprises the aqueous phase component; the aqueous phase component comprises: 1-2.25% of an osmotic pressure regulator, 0.01-1% of an auxiliary emulsifier, and the balance of water; the amount of water is such that the total mass percentage of the oil phase component and the aqueous phase component is 100%.

[0044] In the present invention, the mass percentage of the osmotic pressure regulator is 1-2.25%, preferably 1.32-2.0%. The osmotic pressure regulator preferably includes one or more of glycerol, sodium chloride, glucose, phosphate, and citrate. The present invention does not specifically limit the specific types of the phosphate and citrate; the corresponding phosphates and citrates well known in the art can be used. When the osmotic pressure regulator is two or more of the above, the present invention does not specifically limit the ratio of the different types of osmotic pressure regulators, and can be adjusted according to actual needs.

[0045] In the present invention, the weight percentage of the auxiliary emulsifier is 0.01-1%, preferably 0.03-0.5%; the auxiliary emulsifier preferably includes one or more of oleic acid, sodium oleate, and phosphatidylglycerol. When the auxiliary emulsifier is two or more of the above, the present invention does not specifically limit the ratio of different types of auxiliary emulsifiers and can be adjusted according to actual needs.

[0046] The present invention provides a method for preparing the rolapitant fat emulsion injection described in the above technical solution, comprising the following steps:

[0047] mixing rolapitant and a cosolvent, and mixing the resulting mixture with an emulsifier and vegetable oil to obtain an oil phase;

[0048] mixing water, an osmotic pressure regulator, and a co-emulsifier to obtain an aqueous phase;

[0049] The aqueous phase and the oil phase are combined under shear conditions to obtain colostrum;

[0050] The colostrum is sequentially subjected to high-pressure homogenization and sterilization to obtain rolapitant fat emulsion injection.

[0051] The present invention has no particular limitation on the “mixing” involved in the preparation method, and the mixture may be stirred and mixed uniformly according to a process well known in the art.

[0052] In the present invention, the shearing rotation speed is preferably 10,000 rpm; the pressure of the high-pressure homogenization is 10,000 to 20,000 psi, and the number of times is preferably 5 times.

[0053] In the present invention, the sterilization temperature is preferably 121° C., and the sterilization time is preferably 15 minutes.

[0054] The present invention provides the use of the rolapitant fat emulsion injection described in the above technical solution or the rolapitant fat emulsion injection prepared by the preparation method described in the above technical solution in the preparation of an anti-nausea and vomiting drug. The present invention does not specifically limit the method of application, and the application can be carried out according to methods well known in the art.

[0055] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0056] Example 1

[0057] (1) According to the prescription in Table 1, accurately weigh the prescribed amount of rolapitant hydrochloride monohydrate, and stir and dissolve it with the prescribed amount of anhydrous ethanol at room temperature to obtain a rolapitant ethanol solution; accurately weigh the prescribed amount of egg yolk lecithin, stir and mix with the prescribed amount of soybean oil and medium-chain triglycerides, and inject the rolapitant ethanol solution into the obtained oil phase mixture, stir and dissolve until it becomes clear and transparent, to obtain an oil phase;

[0058] (2) Weigh the prescribed amount of purified water, glycerol, and sodium oleate, and dissolve them under magnetic stirring until they are completely dissolved to obtain an aqueous phase;

[0059] (3) slowly adding the aqueous phase to the oil phase for merging, and after merging, shearing at 10,000 rpm and homogenizing the resulting colostrum at 10,000 psi for 5 times;

[0060] (4) Sterilize at 121°C for 15 min to obtain rolapitant fat emulsion injection.

[0061] Table 1 Prescription composition of Example 1 Rolapitant fat emulsion injection (Preparation 1)

[0062]

[0063]

[0064] Example 2

[0065] According to the prescription in Table 2, the amounts of glycerol (for injection) and sodium oleate were changed. Preparation 2 was homogenized 5 times at 20,000 psi, and Preparation 3 was homogenized 5 times at 10,000 psi. The rest of the prescription and process were referred to Example 1.

[0066] Table 2 Example 2 Rolapitant fat emulsion injection prescription composition

[0067]

[0068] Example 3

[0069] (1) According to the prescription in Table 3, the processes of Preparation 4 and Preparation 5 are the same. The prescribed amount of rolapitant hydrochloride monohydrate is accurately weighed and dissolved in the prescribed amount of anhydrous ethanol at room temperature by stirring to obtain a rolapitant ethanol solution; the prescribed amount of egg yolk lecithin is accurately weighed and dissolved in the prescribed amount of medium-chain triglycerides (or soybean oil) in a water bath, and the rolapitant ethanol solution is injected into the obtained oil phase mixture, stirred and dissolved until it becomes clear and transparent, to obtain an oil phase;

[0070] (2) Weigh the prescribed amount of purified water, glycerol, and sodium oleate, and dissolve them under magnetic stirring until they are completely dissolved to obtain an aqueous phase;

[0071] (3) slowly adding the aqueous phase to the oil phase for merging, and after merging, shearing at 10,000 rpm and homogenizing the resulting colostrum at 20,000 psi for 5 times;

[0072] (4) Sterilize at 121°C for 15 min to obtain rolapitant fat emulsion injection.

[0073] Table 3 Prescription composition of Rolapitant fat emulsion injection in Example 3

[0074]

[0075] Example 4

[0076] According to the prescription in Table 4, the dosage was adjusted to increase the amount of rolapitant hydrochloride monohydrate. The preparation process of Preparation 6 was the same as that of Example 1.

[0077] Table 4 Prescription composition of Example 4 Rolapitant fat emulsion injection (Preparation 6)

[0078]

[0079]

[0080] Example 5

[0081] (1) According to the prescription in Table 5, accurately weigh the prescribed amount of rolapitant hydrochloride monohydrate, and stir and dissolve it with the prescribed amount of anhydrous ethanol at room temperature to obtain a rolapitant ethanol solution; accurately weigh the prescribed amount of egg yolk lecithin, stir and mix it with the prescribed amount of medium-chain triglycerides, and inject the rolapitant ethanol solution into the obtained oil phase mixture, stir and dissolve it until it becomes clear and transparent, to obtain an oil phase;

[0082] (2) Weigh the prescribed amount of purified water, glycerol, and oleic acid, and dissolve them under magnetic stirring until they are completely dissolved to obtain an aqueous phase;

[0083] (3) slowly adding the aqueous phase to the oil phase for merging, shearing at 10,000 rpm after merging, adjusting the pH value of the colostrum to 8.50 with sodium hydroxide solution, and homogenizing the resulting colostrum at 20,000 psi for 5 times;

[0084] (4) Sterilize at 121°C for 15 min to obtain rolapitant fat emulsion injection.

[0085] Table 5 Prescription composition of Example 5 Rolapitant fat emulsion injection (Preparation 7)

[0086]

[0087] Comparative Example 1

[0088] (1) According to the prescription in Table 6, accurately weigh the prescribed amount of rolapitant acid monohydrate, HS15, soybean oil and medium-chain triglycerides, stir and dissolve them completely; then add the prescribed amount of rolapitant hydrochloride monohydrate, mix well to form an oil phase suspension;

[0089] (2) Weigh the prescribed amount of anhydrous sodium dihydrogen phosphate, sodium chloride, and purified water, and dissolve them under magnetic stirring at room temperature until they are completely dissolved to obtain the aqueous phase;

[0090] (3) The aqueous phase was slowly added to the oil phase to form colostrum. After the colostrum was formed, the colostrum was sheared at 10,000 rpm for 10 minutes and homogenized at 10,000 psi for 5 times.

[0091] (4) Filter the sample through a 0.45 μm PES filter membrane to obtain rolapitant hydrochloride intravenous injection emulsion.

[0092] Table 6 Prescription composition of Comparative Example 1 (Preparation 8, commercially available prescription)

[0093]

[0094] Performance Testing

[0095] The samples prepared in different cases were placed at 25°C (long-term), 40°C (accelerated) and 60°C (high temperature) to investigate the stability of the samples. Sampling was carried out to test the stability at high temperature for 5 days, 10 days, 30 days, accelerated for 1M, 2M, 3M and long-term for 3M. The test methods were General Rules 0982, General Rules 0631, General Rules 0632 and General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The results are shown in Tables 7-8.

[0096] Table 7 Stability of samples of Example 1 (Formulation 1)

[0097]

[0098]

[0099] As shown in Table 7, the sample of Preparation 1 was relatively stable under long-term conditions, with no significant changes in particle size, Zeta potential, pH value, osmotic pressure and content, and a slight increase in total impurities.

[0100] Table 8 Physicochemical stability of samples from Example 2 (Formulation 2 and Formulation 3)

[0101]

[0102] From the data of Formulation 2 in Table 8, it can be seen that increasing the homogenization pressure reduces the particle size of the sample; increasing the amount of glycerol increases the osmotic pressure molar concentration.

[0103] From the data of Preparation 3 in Table 8, it can be seen that when the mass percentage of sodium oleate is 0.01%, the absolute value of the Zeta potential of the sample decreases.

[0104] Test Case

[0105] 1) The pharmacokinetic characteristics of rolapitant fat emulsion were investigated in SD rats after a single intravenous injection. The concentration of the two test products (Formulation 1 and Formulation 8) was set at 2 mg / mL and the administration volume was 10 mL / kg, resulting in a dose of 20 mg / kg. The specific experimental design is shown in Table 9:

[0106] Table 9 Experimental design table

[0107]

[0108] Pharmacokinetic testing:

[0109] Sampling animals and time points: All experimental animals were sampled before administration (Pre), and at 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, and 24 hours after administration; time error was allowed: ±2 minutes for <2 hours, ±10 minutes for ≤2 hours to 8 hours, and ±30 minutes for 24 hours;

[0110] Sampling requirements: Collect approximately 0.2 mL / sample / time from the jugular vein or other appropriate sites, anticoagulant with EDTA-K2. After collection, gently invert several times to ensure that the sample and anticoagulant are fully mixed. Store in an ice box and then transport as soon as possible.

[0111] Sample pretreatment: Centrifuge at room temperature, approximately 4000 rpm, for 10 minutes, and freeze for temporary storage after separation.

[0112] Detection and processing: LC-MS / MS instrumentation was used for analysis. Samples with concentrations above the upper limit of quantification should be diluted with the corresponding blank matrix and re-measured. When calculating pharmacokinetic parameters, samples with concentrations below the lower limit of quantification should be marked as below the lower limit of quantification (BLQ) to minimize the impact of zero values ​​on AUC calculations. Conventional pharmacokinetic parameters were calculated using Phoenix WinNonlin software.

[0113] Test results such as Figures 1-2 and Table 10:

[0114] Figure 1 The drug concentration-time curves of Rolapitant injection prepared in Example 1 (self-made group) and Comparative Example 1 (control group);

[0115] Figure 2 The semi-logarithmic curves of the rolapitant injection prepared in Example 1 (self-made group) and Comparative Example 1 (control group) are shown.

[0116] Table 10 Comparison results of formulation 1 and formulation 8

[0117]

[0118] Depend on Figures 1-2 As shown in Table 10, after intravenous injection of 20 mpk of ropitant hydrochloride injection (Formulation 1) in male rats, the clearance (CL) was 15.63 mL / min / kg and the AUClast was 20300.15 ng / mL*h. After intravenous injection of 20 mpk of ropitant hydrochloride injection (control group) in male rats, the clearance (CL) was 8.60 mL / min / kg and the AUClast was 35092.68 ng / mL*h. This indicates that there was no significant difference in the pharmacokinetics of Formulation 1 and the control group (Formulation 8) in SD rats, and the two can be considered PK equivalent.

[0119] 2) Hemolytic studies

[0120] The concentration of both test products was 2 mg / mL. A negative control group (administered 0.9% sodium chloride injection) and a positive control group (administered sterile water for injection) were set up as experimental controls. The specific experimental design is shown in Table 11:

[0121] Table 11 Design of in vitro hemolysis test of New Zealand rabbit erythrocytes

[0122]

[0123]

[0124] Rabbit blood collection and preparation of 2% red blood cell suspension

[0125] 10 mL of blood was collected from a New Zealand rabbit heart into a clean glass bottle containing a stirrer and then slowly shaken for 10 minutes to remove fibrinogen, forming a defibrillated blood. Sodium chloride injection (normal saline) at 10 times the blood volume was added to the tube, followed by centrifugation at 1500 rpm for 10 minutes. After removing the supernatant, the red blood cells were washed twice with 0.9% sodium chloride solution until the supernatant no longer showed red color. The resulting red blood cells were resuspended in 0.9% sodium chloride solution to prepare a 2% (v / v) red blood cell suspension.

[0126] Test tube sample addition

[0127] According to Table 11, take 2 sets (7 tubes / set) of clean glass test tubes (A1-A5 in each set of test tubes are used for Preparation 1, B1-B5 for Preparation 8, A6 / B6 for negative control, and A7 / B7 for positive control). Add 2% red blood cell suspension, 0.9% sodium chloride solution or sterile water for injection, and 2 test sample preparations in the order shown in Table 10. Gently shake the test tubes manually until mixed. Immediately incubate the test tubes at 37±1°C.

[0128] The results are as follows Figures 3-4 (Preparation 1) and Figures 5-6 As shown in (Preparation 8), Figures 3 to 6 In the figure, the test tubes represent A1 / B1, A2 / B2, A3 / B3, A4 / B4, A5 / B5, A6 / B6, and A7 / B7 from left to right. The specific results are:

[0129] Before incubation: Red blood cells are evenly distributed in all tubes. The liquid in tubes A1-A5 is opaque pink, in tubes A6 and A7 is clear red, and in tubes B1-B7 is clear red.

[0130] Incubate for 15 minutes: Red blood cells in tubes A1-A6 and B1-B6 begin to settle. The lower layer of tubes A1-A5 is pink and the upper layer is milky white. The upper layer of tubes A6 is transparent and colorless, and A7 is clear red. The upper layer of tubes B1-B6 is transparent and colorless, and A7 is clear red.

[0131] Incubate for 30 minutes to 3 hours: The red blood cells in tubes A1-A6 and B1-B6 gradually settle completely. The lower layer in tubes A1-A5 is pink, the upper layer is milky white, and A7 is clear red. The lower layer in tubes B1-B6 is clear dark red, the upper layer is transparent and colorless, and A7 is clear red.

[0132] After incubation and mixing, red blood cells are evenly distributed in tubes A1-A6 and B1-B6. Tubes A1-A5 are opaque pink, A6 is uniform red, A7 is clear red, B1-B5 are uniform dark red, B6 is uniform red, and B7 is clear red.

[0133] No hemolysis or aggregation occurred in the negative control tube solution, and all the solutions in the positive control tube were hemolyzed, and it was determined that the test system was established. When the erythrocyte suspension to which Rolapitant Hydrochloride Injection (Preparation 1) was added was incubated, the erythrocytes settled, and the upper liquid was milky white (suspected to be due to the color interference of the test sample itself), and no erythrocyte aggregation was observed, indicating that no hemolysis or erythrocyte aggregation occurred. When the erythrocyte suspension to which Rolapitant Hydrochloride Injection (Comparative Example 1) was added was incubated, the erythrocytes settled, and the upper liquid was transparent and colorless, and no erythrocyte aggregation was observed, indicating that no hemolysis or erythrocyte aggregation occurred.

[0134] In summary, under the experimental conditions, no hemolysis or aggregation was observed in the 2% red blood cell suspension of New Zealand rabbits with rolapitant hydrochloride injection (Preparation 1) and rolapitant hydrochloride injection (Preparation 8) at a concentration of 2 mg / mL. The test results were judged to be negative, indicating that this product can be used for intravenous injection without the risk of hemolysis.

[0135] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A rolapitant fat emulsion injection, characterized in that: comprising an oil phase component and a water phase component; The oil phase components include: 0.1-1% of rolapitant, 0.1-5% of cosolvent, 5-10% of vegetable oil, and 1-2% of emulsifier; The aqueous phase components include: 1-2.25% osmotic pressure regulator, 0.01-1% auxiliary emulsifier, and the balance water; The amount of water used is such that the total mass percentage of the oil phase component and the water phase component is 100%.

2. The rolapitant fat emulsion injection according to claim 1, characterized in that: The co-solvent includes one or more of ethanol, polyethylene glycol, glycerol and ethylene glycol.

3. The rolapitant fat emulsion injection according to claim 1, characterized in that: The vegetable oil includes one or more of soybean oil, medium-chain oil, olive oil, castor oil, sesame oil and hydrogenated vegetable oil.

4. The rolapitant fat emulsion injection according to claim 1, characterized in that: The emulsifier includes one or more of egg yolk lecithin, egg yolk phosphatidylcholine, soybean lecithin, polyene phosphatidylcholine and hydrogenated phospholipids.

5. The rolapitant fat emulsion injection according to claim 1, characterized in that: The osmotic pressure regulator includes one or more of glycerol, sodium chloride, glucose, phosphate and citrate.

6. The rolapitant fat emulsion injection according to claim 1, characterized in that: The auxiliary emulsifier includes one or more of oleic acid, sodium oleate and phosphatidylglycerol.

7. The method for preparing the rolapitant fat emulsion injection according to any one of claims 1 to 6, characterized in that: The following steps are involved: mixing rolapitant and a cosolvent, and mixing the resulting mixture with an emulsifier and vegetable oil to obtain an oil phase; mixing water, an osmotic pressure regulator and an auxiliary emulsifier to obtain an aqueous phase; The water phase and the oil phase are combined under shear conditions to obtain colostrum; The colostrum is sequentially subjected to high pressure homogenization and sterilization to obtain rolapitant fat emulsion injection.

8. The preparation method according to claim 7, characterized in that: The pressure of the high-pressure homogenization is 10000-20000 psi.

9. Use of the rolapitant fat emulsion injection according to any one of claims 1 to 6 or the rolapitant fat emulsion injection prepared by the preparation method according to any one of claims 7 to 8 in the preparation of anti-nausea and vomiting drugs.

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