Lyophilized compositions containing anti-cd70-drug conjugates, lyophilized formulations, methods of making and uses thereof

Abstract

The application provides a freeze-dried composition containing an anti-CD70-drug conjugate, which comprises ARX305 and pharmaceutically acceptable buffers, surfactants and freeze-drying protectants, wherein the pH value of the freeze-dried composition is 5.5-6.0. The application also provides a freeze-dried preparation containing an anti-CD70-drug conjugate and a preparation method thereof. The application also provides the use of the freeze-dried composition and the freeze-dried preparation. The freeze-dried composition and the freeze-dried preparation provided by the application have excellent product performance, can be used for treating various cancers, and the preparation method is simple, easy to operate, high in production efficiency, and has very important economic and social values.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, specifically to a lyophilized composition containing an anti-CD70-drug conjugate, a lyophilized formulation obtained from the lyophilized composition, a method for preparing the same, and its uses. Background Technology

[0002] CD70 is a type II transmembrane protein, a trimer composed of 193 amino acids, including an extracellular binding region, a transmembrane region, a short intracellular sequence, and two N-terminal glycosylated sequences. It is a member of the tumor necrosis factor receptor (TNFR) superfamily. CD70 and its receptor CD27 play important roles in regulating T cell and B cell activation, proliferation, and differentiation, and in maintaining the body's immune response. CD70 protein expression and CD70 gene amplification are associated with poor clinical prognosis. Studies have shown that CD70 is expressed in both hematologic malignancies and solid tumors, with varying positivity rates among different cancer types. Currently, CD70 has become a potential target for anti-tumor drugs, and several CD70-targeted drugs are in clinical trials. For patients with CD70-positive tumors, treatment with CD70-targeted drugs holds promise for improving their quality of life and prolonging their survival.

[0003] Antibody-drug conjugates (ADCs) are a class of anti-tumor drugs composed of monoclonal antibodies linked to small cytotoxic drugs via linkers. They combine the high targeting ability of monoclonal antibodies with the potent cytotoxicity of small molecule drugs, resulting in better efficacy and fewer side effects, making them a popular area of ​​research in cancer treatment. Compared to conventional biologics, ADCs have more complex structures and more unstable factors: First, the conjugated small cytotoxic drugs may detach during long-term storage, leading to a decrease in the drug-antibody ratio and an increase in the content of free small molecules; second, the naked antibody portion of the ADC may undergo physical degradation such as aggregation, precipitation, or adsorption, as well as chemical degradation such as deamidation, oxidation, hydrolysis, changes in charge heterogeneity, and fragmentation; third, small molecule drugs are usually hydrophobic, and the ADC as a whole may be more prone to aggregation than the unconjugated monoclonal antibody, forming particles and surface adsorption. Therefore, to provide stable ADCs during storage and transportation, careful selection of the dosage form, carrier, pH value, buffer system, excipients and / or stabilizers, and preparation process is essential.

[0004] Recombinant humanized anti-CD70 monoclonal antibody-AS269 conjugate (ARX305) is a new generation of monoclonal antibody-drug conjugates used to treat advanced tumors expressing CD70. Its structural formula is as follows: Figure 1As shown, the original anti-CD70 monoclonal antibody's heavy chain amino acid at position 119 was mutated to an acetylphenylalanine residue using codon expansion technology. This residue was then oxime-treated with a small molecule toxin containing a hydroxylamine group, resulting in site-directed conjugation to obtain a conjugate with a major drug-toxin ratio of 1:2 (Chinese Patent CN 104619351B provides a detailed description of this process, specifically the amino acid and nucleotide sequences of CD70-7F2-AS269 described in SEQ ID NO.1–SEQ ID NO.3 and SEQ ID NO.5–SEQ ID NO.9. The disclosure of CN104619351B is incorporated herein by reference). Regarding ARX305, no formulation-related technologies have been reported to date. Summary of the Invention

[0005] To fill the gap in the existing technology related to ARX305 formulations, one object of the present invention is to provide a lyophilized composition containing an anti-CD70 drug conjugate, which has excellent stability.

[0006] Another object of the present invention is to provide a lyophilized formulation containing an anti-CD70-drug conjugate and a method for preparing the same.

[0007] Another object of the present invention is to provide the use of the lyophilized composition and the lyophilized formulation.

[0008] The lyophilized composition containing an anti-CD70 drug conjugate provided by the present invention comprises: ARX305 and pharmaceutically acceptable buffer, surfactant and lyophilization protectant, wherein the pH value of the lyophilized composition is 5.5 to 6.0.

[0009] Through stability studies at different pH values, the inventors of this invention discovered that at lower pH values, the antibody-drug conjugate is more prone to the shedding of the small molecule drug AS269, while at higher pH values, a decrease in the CEX main peak and an increase in acidic and basic peaks are more likely to occur. The resulting lyophilized composition achieves optimal stability within a pH range of 5.5 to 6.0. The pH values ​​of the lyophilized composition include, but are not limited to, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, or about 6.0. In some preferred embodiments, the pH value of the lyophilized composition is 5.6 to 5.8, for example, pH 5.7.

[0010] The pharmaceutically acceptable buffers, surfactants, and lyophilization protectants described in this invention can be common types in the pharmaceutical field, especially those commonly used in ADC formulations. In some preferred embodiments, the buffers include, but are not limited to, histidine-histidine hydrochloride buffer, acetate-sodium acetate buffer, citrate-sodium citrate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, etc.; the surfactants include, but are not limited to, polysorbate 20, polysorbate 80, etc.; and the lyophilization protectants include, but are not limited to, sucrose, trehalose, etc.

[0011] In some preferred embodiments, the lyophilized composition comprises 10–20 mg / ml of ARX305, 15–25 mM of buffer solution (total buffer salt concentration), 0.2–0.6 mg / ml of surfactant, and 60–100 mg / ml of lyophilization protectant. In this case, the pH of the lyophilized composition is 5.5–6.0, preferably 5.6–5.8, for example 5.7.

[0012] In some preferred embodiments, the lyophilized composition comprises 10–20 mg / ml of ARX305, 15–25 mM of histidine-histidine hydrochloride buffer, 0.2–0.6 mg / ml of polysorbate 80, and 60–100 mg / ml of trehalose. In this case, the pH of the lyophilized composition is 5.5–6.0, preferably 5.6–5.8, for example 5.7.

[0013] In some further preferred embodiments, the freeze-dried composition comprises any one of the following compositions 1)-6):

[0014] Composition 1): 10 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.2 mg / ml polysorbate 80 and 80 mg / ml trehalose;

[0015] Composition 2): 10 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.4 mg / ml polysorbate 80 and 80 mg / ml trehalose;

[0016] Composition 3): 10 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.6 mg / ml polysorbate 80 and 80 mg / ml trehalose;

[0017] Composition 4): 20 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.2 mg / ml polysorbate 80 and 80 mg / ml trehalose;

[0018] Composition 5): 20 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.4 mg / ml polysorbate 80, and 80 mg / ml trehalose; and

[0019] Composition 6): 20 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.6 mg / ml polysorbate 80 and 80 mg / ml trehalose.

[0020] The structure of the ARX305 described in this invention is as follows: Figure 1 As shown in SEQ ID NO:1, the heavy chain amino acid sequence of the anti-CD70 monoclonal antibody represents an acetylphenylalanine residue at position 119. The light chain amino acid sequence of the anti-CD70 monoclonal antibody is shown in SEQ ID NO:2. A small molecule toxin containing a hydroxylamine group undergoes oxime reaction with the acetylphenylalanine residue at position 119 of the heavy chain of the anti-CD70 monoclonal antibody, resulting in site-directed coupling and yielding the conjugate ARX305 with a major drug-to-antibody ratio of 1:2.

[0021]

[0022] The present invention also provides a lyophilized formulation containing an anti-CD70-drug conjugate, which is prepared from the lyophilized composition described in any one of the above technical solutions.

[0023] The present invention also provides a method for preparing the lyophilized formulation containing the anti-CD70-drug conjugate, which is obtained by sequentially subjecting the lyophilized composition described in any of the above technical solutions to pre-freezing, a first sublimation drying, and a second sublimation drying, wherein the pre-freezing process is subjected to annealing treatment, and the resistivity of the lyophilized composition at the annealing temperature is 10 to 30% (including but not limited to about 10%, about 12%, about 15%, about 18%, about 20%, about 22%, about 25%, about 28%, or about 30%) of the resistivity of the lyophilized composition at the pre-freezing temperature.

[0024] The inventors of this invention discovered that resistivity reflects the freezing state of the lyophilized composition during the pre-freezing process. Typically, the resistivity is approximately 0% in the solution state and approaches 100% after complete freezing. At different annealing temperatures, varying degrees of ice crystal melting occur, resulting in different degrees of resistivity decrease. Lower annealing temperatures result in less ice crystal melting and a smaller decrease in resistivity, while higher annealing temperatures lead to more ice crystal melting and a greater decrease in resistivity. The preparation method provided by this invention, by controlling the degree of ice crystal melting (reflected in the change in resistivity), can control and improve the microstructure of the ice crystals, forming loose channels within them. This not only improves the efficiency of subsequent sublimation drying but also significantly improves the appearance, reconstitution effect, moisture content, and other properties of the resulting lyophilized formulation, and also greatly increases the yield of the finished product.

[0025] In some preferred embodiments, the pre-freezing temperature is -50 to -40°C, and the pre-freezing time is 5 to 12 hours. In this case, the annealing temperature can be determined accordingly based on the pre-freezing temperature and the aforementioned change in resistivity (e.g., -3 to -10°C), and the annealing time is 2 to 6 hours. For example, when the pre-freezing temperature is -45°C, the annealing temperature can be -5 to -9°C.

[0026] In some preferred embodiments, the pre-freezing includes the following process: cooling the freeze-dried composition (e.g., within 0.5 to 1.5 h) to 3 to 8 °C and holding it at that temperature for 0.5 to 1.5 h (e.g., within 20 to 50 min), then cooling it to -50 to -40 °C and holding it at that temperature for 1 to 3 h, then annealing it at the annealing temperature (e.g., within 20 to 50 min) for 2 to 6 h (e.g., within 20 to 50 min), then cooling it again to -50 to -40 °C and holding it at that temperature for 3 to 8 h.

[0027] In some further preferred embodiments, the pre-freezing includes the following process: cooling the freeze-dried composition (e.g., over 0.5 to 1.5 hours) to 4 to 6°C and holding it at that temperature for 0.8 to 1.2 hours (e.g., over 20 to 50 minutes), then cooling it to -47 to -43°C and holding it at that temperature for 1.8 to 2.5 hours, followed by annealing at -5 to -9°C (e.g., over 20 to 50 minutes), and then cooling it again to -47 to -43°C and holding it at that temperature for 4 to 6 hours (e.g., over 20 to 50 minutes). For example, the pre-freezing includes the following process: cooling the freeze-dried composition to 5°C and holding it at that temperature for 1 hour, then cooling it to -45°C and holding it at that temperature for 2 hours, then annealing it at -5 to -9°C for 3 hours, and then cooling it again to -45°C and holding it at that temperature for 5 hours.

[0028] In some preferred embodiments, the temperature for the first sublimation drying is -23 to -18°C, the vacuum degree is 10 to 15 Pa, and the holding time is 30 to 50 hours. In some more preferred embodiments, the first sublimation drying includes the following process: heating the pre-frozen product of the lyophilized composition to -23 to -18°C (preferably -20°C) within 3 to 6 hours, and holding it at this temperature and under a vacuum degree of 10 to 15 Pa (preferably 13 Pa) for 30 to 50 hours (preferably 40 hours).

[0029] In some preferred embodiments, the secondary sublimation drying is performed at a temperature of 35–40°C, a vacuum of 10–15 Pa, and a holding time of 10–18 h. In some more preferred embodiments, the secondary sublimation drying includes the following process: the lyophilized composition, after pre-freezing and primary sublimation drying, is heated to 35–40°C (preferably 37°C) within 3–6 h, and held at this temperature and a vacuum of 10–15 Pa (preferably 13 Pa) for 10–18 h (preferably 12 h).

[0030] The present invention also provides the use of the lyophilized composition or the lyophilized preparation described in any of the above technical solutions in the preparation of a medicament for treating cancer.

[0031] In the uses described in this invention, the cancers include common solid tumors and hematologic malignancies, including but not limited to renal cell carcinoma (especially advanced renal cell carcinoma), nasopharyngeal carcinoma, ovarian cancer, glioma, hepatocellular carcinoma, colorectal cancer, lung cancer, prostate cancer, biliary tract cancer, relapsed / refractory diffuse large B-cell lymphoma, relapsed / refractory follicular lymphoma, relapsed / refractory Hodgkin lymphoma, and relapsed / refractory acute myeloid leukemia.

[0032] The technical solution provided by this invention has the following advantages:

[0033] The lyophilized composition provided by this invention is designed for the novel ADC drug ARX305. By setting specific pH values, composition formulations, etc., it can effectively reduce the drop of small molecule drugs and the decline of the CEX main peak. The resulting lyophilized product has excellent stability and can be stored stably for a long time.

[0034] The freeze-dried formulation provided by this invention employs an annealing process controlled by resistivity during preparation. The resulting freeze-dried formulation product not only has excellent appearance and low moisture content, but also good reconstitution effect (short reconstitution time and clear solution after reconstitution), and high product yield, which greatly reduces production costs and improves production efficiency.

[0035] In summary, the lyophilized compositions and lyophilized formulations provided by this invention have excellent product performance, can be used to treat a variety of cancers, and are simple to prepare, highly operable, and have high production efficiency, thus possessing significant economic and social value. Attached Figure Description

[0036] Figure 1 This is a schematic diagram of the structure of the ADC drug ARX305, in which an antibody protein is coupled to two small molecule compounds.

[0037] Figure 2 Typical appearance images of lyophilized formulation samples prepared under different processes in Example 3 (left: intact appearance; right: surface peeling / bottom shrinkage). Detailed Implementation

[0038] The technical solution of the present invention will be further described in detail below with reference to specific embodiments.

[0039] The ARX305 (anti-CD70-ADC) and ADC reference (RS) used in the examples were obtained from Zhejiang Xinma Biomedical Co., Ltd., and other reagents or raw materials were commercially available products unless otherwise specified.

[0040] The detection items and detection conditions used in the examples are as follows:

[0041] Appearance: Visually inspected using a clarity meter.

[0042] Protein concentration: Detected using a Cary 100 UV-Vis spectrophotometer.

[0043] pH value: Mettler Micro Pro pH meter detection.

[0044] Moisture content: Measured using a Mettler Toledo C30D Karl Fischer moisture analyzer.

[0045] Turbidity: Absorption at UV340 nm was detected using Nano Drop 2000.

[0046] Cation exchange chromatography (CEX-HPLC):

[0047] The acidic peak, main peak, and basic peak in the sample were determined using an Agilent 1260 HPLC instrument, and their percentage content was calculated using the area normalization method. The chromatographic conditions are as follows:

[0048]

[0049]

[0050] AS269 (small molecule toxin) residue:

[0051] The AS269 residue in the sample was determined by HPLC under the following chromatographic conditions:

[0052]

[0053] Example 1: Stability Study at Different pH Values

[0054] Design formulations with different pH values ​​1-10 (see Table 1).

[0055] Table 1 Formulations for different pH values

[0056]

[0057]

[0058] The formulation sample was prepared as follows:

[0059] (1) Prepare buffer solutions according to the composition shown in Table 1, and adjust the pH value to the target value using hydrochloric acid or sodium hydroxide if necessary;

[0060] (2) Prepare concentrated excipient solutions using buffer solutions, with polysorbate 80 and sucrose concentrations of 14 mg / ml (70 times the target concentration) and 800 mg / ml (10 times the target concentration), respectively.

[0061] (3) Take 10 portions of ADC solution and add them to pre-treated ultrafiltration centrifuge tubes (model: 30k, 15ml). Centrifuge for 20 min, add buffer solution and centrifuge for 20 min to change the solution. Repeat the solution change and concentration operation four times to obtain concentrated ADC solution containing target buffer (concentration of approximately 20mg / ml).

[0062] (4) Measure the protein concentration of the ADC solution after the solution is concentrated and add the corresponding amount of excipient concentrated solution according to the calculated value, and use buffer solution to make up to the target volume;

[0063] (5) The obtained sample was filtered through a 0.22μm sterile filter, dispensed (6ml), and then capped and crimped.

[0064] Stability assessment:

[0065] The samples were placed at 25℃ for two months for observation. The results are shown in Table 2, where T0 represents the time point when the samples were first placed, 25℃-1M represents the time point after one month of sample placement, and 25℃-2M represents the time point after two months of sample placement.

[0066] Table 2 Summary of the stability study results of the formulation at different pH values

[0067]

[0068]

[0069] Results analysis:

[0070] The AS269 residue results show that, under 25°C conditions, formulations with pH < 5.5 all exhibited an increasing trend in AS269 residue. The lower the pH, the higher the AS269 residue, indicating that small molecules are more likely to detach under pH conditions less than 5.5.

[0071] The CEX results show that the CEX main peak decreases and the acidic and alkaline peaks increase more easily when the pH value is greater than 6.0.

[0072] Therefore, the pH value of the freeze-dried composition is crucial to the product stability. The pH value of the freeze-dried composition of the present invention is preferably between 5.5 and 6.0, and more preferably around 5.7.

[0073] Example 2: Formulation stability study at different ADC concentrations and polysorbate 80 (PS80) concentrations

[0074] Six formulations were prepared by setting two gradients of ADC concentration (10 mg / ml and 20 mg / ml) and three gradients of polysorbate 80 concentration (0.2 mg / ml, 0.4 mg / ml, and 0.6 mg / ml) (Table 3). The prepared samples were subjected to shaking at 300 rpm at 25°C for 1 day and 3 days, followed by three and five freeze-thaw cycles (freezing at -80°C for 6 hours, then thawing at 25°C for 2 hours). The results (Table 4) showed that the samples in formulations 11-16 did not exhibit significant changes in any of the indicators before and after shaking and freeze-thaw cycles. Therefore, under conditions of approximately pH 5.7, the protein stability was good within the range of 10-20 mg / ml protein concentration and 0.2-0.6 mg / ml polysorbate 80 concentration.

[0075] Table 3 Summary of formulations screened by ADC concentration and PS80 concentration

[0076]

[0077] Table 4. Results of stability studies using ADC and PS80 concentrations.

[0078]

[0079]

[0080] Example 3: Study on freeze-drying process

[0081] 1. Investigation and optimization of the platform's freeze-drying process

[0082] As shown in Example 2, the formulation showed good stability when the concentration of anti-CD70-ADC was 10-20 mg / ml and the concentration of PS80 was 0.2-0.6 mg / ml. Given that the low concentration of PS80 had achieved good stability and that the lower concentration of protein had a lower stability risk, the concentrations of ADC and PS80 were determined to be 10 mg / ml and 0.2 mg / ml, respectively, for the study of the freeze-drying process.

[0083] Formula 11, consisting of 10 mg / ml anti-CD70-ADC (ARX305), 20 mM histidine-histidine hydrochloride buffer, 80 mg / ml sucrose, and 0.2 mg / ml polysorbate 80, was used to investigate the lyophilization process (all lyophilizer plates had an area of ​​0.1 m²). 2 Each lyophilization process involved placing 20 vials of ADC formulation, each containing 6 ml, with the remainder filled with placebo. The lyophilizer used was a ChristALPHA 2-4LSC lyophilizer from Germany.

[0084] As shown in Table 5, when freeze-dried using the platform process (process-1), a certain proportion of samples exhibited defects such as surface peeling / bottom shrinkage, collapse, and remelting (e.g., Figure 2 (As shown on the right), the reconstitution effect was also poor (long reconstitution time and high turbidity value of the solution after reconstitution). Optimization of the primary drying plate temperature and vacuum level (processes 2 to 9) did not significantly improve the freeze-drying effect.

[0085] Table 5 Summary of the investigation and optimization results of the platform freeze-drying process

[0086]

[0087]

[0088] 2. Annealing process optimization

[0089] Formula 11, consisting of 10 mg / ml anti-CD70-ADC (ARX305), 20 mM histidine-histidine hydrochloride buffer, 80 mg / ml sucrose, and 0.2 mg / ml polysorbate 80, was used to lyophilize samples at different annealing temperatures to investigate the relationship between sample resistivity and lyophilization effect during the annealing stage.

[0090] A resistance probe was inserted into the sample, and the resistivity of the sample during the annealing stage was automatically recorded and calculated by the program. Table 6 shows that when the annealing temperature is above -5℃, the resistivity is less than 10%, and a significant amount of ice crystals melt. Compared to process-7, which does not use annealing, the freeze-drying effect (including moisture and resolubilization) is not significantly improved. When the annealing temperature is below -9℃, the resistivity is greater than 30%, and insufficient ice crystal melting also fails to achieve the desired annealing effect. However, when the annealing temperature is within the range of -5℃ to -9℃ (resistivity between 12% and 22%), the freeze-drying effect is significantly improved, and the product appearance is greatly improved (the appearance of products prepared by processes-12 to-14 is as follows). Figure 2 As shown on the left, the appearance is intact, without defects such as peeling or shrinkage. The moisture content has decreased significantly, and the reconstitution time has been greatly shortened from the original 3-6 minutes to 50 seconds or even less than 40 seconds. Moreover, the turbidity after reconstitution has also decreased significantly.

[0091] Table 6 Summary of Annealing Process Optimization Results

[0092]

[0093]

[0094] 3. Process Scale-up

[0095] The optimized process-13 was scaled up on freeze dryers with different plate areas. As shown in Table 7, process-13 exhibits good scale-up performance, yielding high-yield qualified samples on various freeze dryer models, demonstrating good process robustness.

[0096] Table 7 Scale-up Results of Process-13

[0097]

[0098]

[0099] Unless otherwise specified, the terms used in this invention have the meanings commonly understood by those skilled in the art.

[0100] The embodiments described in this invention are for illustrative purposes only and are not intended to limit the scope of protection of this invention. Those skilled in the art can make various other substitutions, changes and improvements within the scope of this invention. Therefore, this invention is not limited to the above embodiments, but is only defined by the claims.

Claims

1. A lyophilized formulation containing an anti-CD70 drug conjugate, characterized in that, The lyophilized formulation is prepared by sequentially subjecting a lyophilized composition to pre-freezing, a first sublimation drying, and a second sublimation drying. The pre-freezing process includes the following steps: cooling the lyophilized composition to 4–6°C and holding it at that temperature for 0.8–1.2 h, then cooling it to -47–-43°C and holding it at that temperature for 1.8–2.5 h, then annealing it at -5–-9°C for 2.5–3.5 h, and finally cooling it again to -47–-43°C and holding it at that temperature for 4–6 h. The resistivity of the lyophilized composition at the annealing temperature is 12–22% of the resistivity of the lyophilized composition at the pre-freezing temperature. The lyophilized composition comprises 10–20 mg / ml ARX305, 15–25 mM histidine-histidine hydrochloride buffer, 0.2–0.6 mg / ml polysorbate 80, and 60–100 mg / ml sucrose, and the pH of the lyophilized composition is 5.

7.

2. The lyophilized formulation according to claim 1, characterized in that, The freeze-dried composition comprises any one of the following compositions 1)-6): Composition 1): 10 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.2 mg / ml polysorbate 80 and 80 mg / ml sucrose; Composition 2): 10 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.4 mg / ml polysorbate 80 and 80 mg / ml sucrose; Composition 3): 10 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.6 mg / ml polysorbate 80 and 80 mg / ml sucrose; Composition 4): 20 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.2 mg / ml polysorbate 80 and 80 mg / ml sucrose; Composition 5): 20 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.4 mg / ml polysorbate 80, and 80 mg / ml sucrose; and Composition 6): 20 mg / ml ARX305, 20 mM histidine-histidine hydrochloride buffer, 0.6 mg / ml polysorbate 80 and 80 mg / ml sucrose.

3. The lyophilized formulation according to claim 1 or 2, characterized in that, The temperature for the first sublimation drying is -23 to -18°C, the vacuum degree is 10 to 15 Pa, and the holding time is 30 to 50 h.

4. The lyophilized formulation according to claim 1 or 2, characterized in that, The secondary sublimation drying process involves a heating temperature of 35–40°C, a vacuum degree of 10–15 Pa, and a holding time of 10–18 h.

5. Use of the lyophilized formulation according to any one of claims 1-4 in the preparation of a medicament for treating advanced renal cell carcinoma.

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