Liquid drug formulation of TACI-FC fusion protein

Abstract

The present invention relates to a liquid pharmaceutical formulation, particularly a liquid pharmaceutical formulation of a TACI-Fc fusion protein, which is a pharmaceutical for treating autoimmune diseases, and which can improve the long-term storage stability of the TACI-Fc fusion protein, maintain its good biological activity, improve pharmaceutical production efficiency, and significantly improve administration convenience.

Description

[Technical Field]

[0001] The present invention relates to a liquid pharmaceutical formulation of TACI-FC fusion protein, which belongs to the field of anti-autoimmune disease medicines. [Background technology]

[0002] The biopharmaceutical market has been growing rapidly in recent years. Fusion protein and antibody drugs have played an important role in the treatment of tumors and autoimmune diseases. However, while they benefit patients, thorough pathological research and technological development have led to the development of new drugs that can undergo physical changes such as aggregation, denaturation, and precipitation, as well as chemical changes such as isomerization, deamidation, and oxidation. Because these changes can affect the safety and efficacy of products, stable formulations are essential to ensure the necessary biological activity before administering fusion protein and antibody drugs to patients. Of the 126 commercially available antibodies approved worldwide from 1986 to February 2021, 10 antibody-drug conjugates, 16 biosimilar drugs, and 3 fusion proteins were included. The above commercially available antibodies include 36 types of lyophilized powders and 100 types of liquid formulations, for a total of 136 products (Reference 1: Robert G. Strickley, et al., A review of formulations of commercially available antibodies. Journal of Pharmaceutical Sciences. 2021(110), 2590-2608.).

[0003] Although lyophilized antibody drug formulations have the advantages of easy long-term storage and stable activity during transportation, the freeze-drying process of lyophilized dosage forms often becomes a bottleneck in pharmaceutical production and leads to increased pharmaceutical production costs. Furthermore, lyophilized antibody formulations must be reconstituted before use, and to minimize aggregation and structural changes of the active ingredient after reconstitution, the reconstitution time is usually extended to ensure effective reconstitution. This increases patient waiting times at hospitals and puts significant pressure on hospitals. Therefore, the development of stable liquid formulations as a preferred option for antibody formulations is increasingly desired.

[0004] However, compared to freeze-dried formulations, the development of liquid antibody formulations that can be stably stored and transported for long periods remains a major challenge in the industry. Liquid antibody formulations are chemically and physically unstable during storage, which may result in the loss of antibody biological activity. In addition, chemical factors such as deamidation, racemization, hydrolysis, oxidation, β-elimination, and disulfide bond exchange, as well as physical factors such as antibody denaturation, aggregation, precipitation, and adsorption, may cause instability of the active ingredient.

[0005] Furthermore, developing highly concentrated liquid antibody drug formulations for subcutaneous or intramuscular injection presents an additional challenge, as high concentrations can significantly increase the viscosity of the liquid formulation and the tendency of the protein to form aggregates, which can contain proteolytic degradation and cause a range of side effects, including problems with undesirable immune responses.

[0006] Antibody-like fusion proteins or antibody-based drugs have unique characteristics compared to conventional polymer drugs. However, the poor stability and complex structure of monoclonal antibody-based drugs present significant challenges in the production and storage of these drugs. Due to the heterogeneous structure of antibodies, particularly the complementarity-determining regions (CDRs) and Fc glycosylation, different monoclonal antibody formulations must be developed on a case-by-case basis. Antibody formulation development presents challenges in terms of antibody conformation, colloidal structure, and chemical structure, including oxidation, isomerization, deamidation, aggregation, denaturation, and cleavage. Exposure to different temperature, humidity, pH, and pressure conditions can alter the antibody's conformation, particularly in the hypervariable region (HVR). Defective products may exhibit reduced activity, and increased immunogenicity can pose significant risks to patients. Therefore, selecting the optimal excipient for antibody protection is crucial (Reference 2: Monoclonal antibodies: formulations of marketed products and recent advances in novel delivery systems, Yanan Cui et al., Drug Development and Industrial Pharmacy, Vol. 43, No. 4, pp. 519-530, 2017). Furthermore, the tendency for fusion protein products to aggregate and suffer from poor stability, leading to adverse immune responses and problems during the purification process, presents a challenging issue to be resolved in the field. The factors that influence fusion protein aggregation are relatively complex and can be divided into external factors, primarily including temperature, physical pressure, and solvent factors (e.g., pH, ionic strength, concentration, and metal ions), and internal factors, primarily including structural features of the fusion protein, sensitive residues, and unpaired cysteines. These factors affect fusion protein aggregation, thereby affecting its stability and shelf life. Therefore, the selection of excipients for fusion proteins requires extensive experimentation and consideration (Reference 3: Production Challenges for Complex Biologics: Fusion Proteins, Stefan R. Schmidt, American Pharmaceutical Review, pp. 1-5, 2017).

[0007] Telitaciccept is a first-in-class recombinant TACI-Fc fusion protein for B cell-mediated autoimmune diseases. It targets and neutralizes two key cell signaling molecules in the B cell pathway, BLyS and APRIL. Telitaciccept is an antibody-like fusion protein for the treatment of human autoimmune diseases, consisting of a truncated TACI and an immunoglobulin Fc with sequence optimization to reduce ADCC and CDC effects. It has demonstrated excellent biological activity and safety, and has been approved by the National Medical Products Administration of China for the treatment of systemic lupus erythematosus. Injectable telitacicept is a lyophilized formulation. However, this formulation significantly limits the drug's production speed, significantly prolongs patient wait times, and increases hospital burdens. Therefore, the development of a liquid formulation is urgently needed. Summary of the Invention [Problem to be solved by the invention]

[0008] The present invention is intended to solve the above problems and provides a liquid pharmaceutical formulation of TACI-Fc fusion protein that effectively enhances the long-term stability of TACI-Fc fusion protein, maintains good biological activity, improves drug production speed and reduces pharmaceutical production costs, significantly improves convenience for patient administration, and better meets the needs of clinical drugs. [Means for solving the problem]

[0009] The liquid formulation of the TACI-Fc fusion protein according to the present invention has excellent stability and can be stable even when stored for 18 months or more under preservative-free, sterile conditions at 4° C. Furthermore, the present inventors surprisingly found that the specific formulation composition developed can also achieve the above requirements even without the use of a surfactant in the formulation composition.

[0010] Specifically, the present invention provides a liquid formulation of a TACI-Fc fusion protein comprising a TACI-Fc fusion protein, a protective agent, a buffer, and a pH adjuster, wherein the TACI-Fc fusion protein comprises (i) the TACI extracellular domain or a fragment thereof that binds to Blys and / or APRIL, and (ii) a fragment of a human immunoglobulin constant domain, the protective agent is hydroxypropyl-β-cyclodextrin, the buffer is succinic acid, and the pH of the liquid formulation is 5.0 to 5.2.

[0011] Furthermore, the concentration of the TACI-Fc fusion protein is in the range of 40 mg / ml to 240 mg / ml.

[0012] Furthermore, the concentration of the TACI-Fc fusion protein is about 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml, 150 mg / ml, 160 mg / ml, 170 mg / ml, 180 mg / ml, 190 mg / ml, 200 mg / ml, 210 mg / ml, 220 mg / ml, 230 mg / ml, or 240 mg / ml.

[0013] Furthermore, the content of the fusion protein was detected by UV-visible spectroscopy, which involves measuring the absorbance of the telitacicept sample at 280 nm, since the protein has a UV absorption maximum at this wavelength. The absorbance at 280 nm, obtained after correcting for the absorbance at 320 nm, is proportional to the protein concentration and follows the Lambert-Beer law to determine the protein content. The formula for calculating the protein content is as follows:

number

[0014] Furthermore, the concentration of the protective agent is in the range of 100 mmol / L to 200 mmol / L. Furthermore, the concentration of the protective agent is about 100 mmol / L, 110 mmol / L, 120 mmol / L, 130 mmol / L, 140 mmol / L, 150 mmol / L, 160 mmol / L, 170 mmol / L, 180 mmol / L, 190 mmol / L, or 200 mmol / L.

[0015] Furthermore, the concentration of the buffering agent is in the range of 5 mmol / L to 15 mmol / L. Furthermore, the concentration of the buffer is about 5 mmol / L, 6 mmol / L, 7 mmol / L, 8 mmol / L, 9 mmol / L, 10 mmol / L, 11 mmol / L, 12 mmol / L, 13 mmol / L, 14 mmol / L, or 15 mmol / L.

[0016] Furthermore, the pH adjuster is an alkaline inorganic salt, more preferably sodium hydroxide. Additionally, the liquid formulation has a pH of about 5.0, 5.1, or 5.2. Preferably, the TACI-Fc fusion protein has the amino acid sequence set forth in SEQ ID NO: 1, and more preferably is telitacicept.

[0017] Sequence number 1 is as follows: SRVDQEERFP QGLWTGVAMR SCPEEQYWDP LLGTCMSCKT ICNHQSQRTC 50 AAFCRSLSCR KEQGKFYDHL LRDCISCASI CGQHPKQCAY FCENKLRSPV 100 NLPPELDKTH TCPPCPAPEA EGAPSVFLFP PKPKDTLMIS RTPEVTCVVV 150 DVSHEDPEVK FNWYVDGVEV HNAKTKPREE QYNSTYRVVS VLTVLHQDWL 200 NGKEYKCKVS NKALPSSIEK TISKAKGQPR EPQVYTLPPS RDELTKNQVS 250 LTCLVKGFYP SDIAVEWESN GQPENNYKTT PPVLDSDGSF FLYSKLTVDK 300 SRWQQGNVFS CSVMHEALHN HYTQKSLSLS PGK 333

[0018] Further, any one of the TACI-Fc-fusion protein liquid formulations contains approximately 10 mmol / L succinic acid, approximately 150 mmol / L hydroxypropyl-β-cyclodextrin, and approximately 80 mg / mL of the TACI-Fc-fusion protein, and the pH of the liquid formulation is approximately 5.1.

[0019] Furthermore, any one of the TACI-Fc-fusion protein liquid formulations contains succinic acid at a concentration of 10 mmol / L, hydroxypropyl-β-cyclodextrin at 150 mmol / L, and TACI-Fc-fusion protein at 80 mg / mL, and the pH of the liquid formulation is about 5.1.

[0020] Additionally, each ml of the liquid formulation of any one of the above TACI-Fc fusion proteins contains about 80 mg of TACI-Fc fusion protein, about 1.18 mg of succinic acid, about 209.85 mg of hydroxypropyl-β-cyclodextrin, and has a pH of about 5.1.

[0021] Furthermore, each ml of a liquid formulation of any one of the above TACI-Fc fusion proteins contains 80 mg of TACI-Fc fusion protein, 1.18 mg of succinic acid, 209.85 mg of hydroxypropyl-β-cyclodextrin, and has a pH of 5.1.

[0022] The invention further relates to the use of any one of the above liquid formulations of TACI-Fc in the manufacture of a medicament for preventing, alleviating and / or treating autoimmune diseases.

[0023] The present invention further relates to a liquid formulation of any one of the above TACI-Fc for use in preventing, alleviating and / or treating autoimmune diseases.

[0024] Furthermore, the present invention relates to a method for preventing, alleviating and / or treating an autoimmune disease, comprising administering to a subject in need thereof any one of the above liquid formulations of TACI-Fc.

[0025] The present invention further relates to a packaged pharmaceutical product comprising any one of the liquid formulations of TACI-Fc. The present invention further relates to a pre-filled syringe containing any one of the above liquid formulations of TACI-Fc, and more preferably, the syringe is an automatic injection pre-filled syringe. [Brief explanation of the drawings]

[0026] [Figure 1] The main effect plot of pH value fitted with the software MODDE is shown. [Figure 2] Main effect plot of arginine concentration fitted with the software MODDE is shown. [Figure 3] The main effect plot of sucrose concentration fitted with the software MODDE is shown. [Figure 4] Main effect plots of protein concentration fitted with the software MODDE are shown. [Figure 5] The contour plots fitted by the software MODDE are shown. The contour plots show the optimal formulations for the DOE experimental design when the formulation does not contain added Arg, the sucrose concentration is in the range of 270 mmol / L to 300 mmol / L, the pH is in the range of 4.95 to 5.1, and the protein concentration is in the range of 80 mg / mL. DETAILED DESCRIPTION OF THE INVENTION

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by one of ordinary skill in the art. For definitions and terminology of the art, see Current Protocols in Molecular Biology (Ausubel).

[0028] The three-letter and one-letter codes for amino acids used in the present invention are as described in J. Biol. Chem, 243, p. 3558 (1968).

[0029] The term "TACI" as used herein is an abbreviation for transmembrane activator and CAML interactor, and is a member of the tumor necrosis factor receptor superfamily. The term "BLys" as used herein refers to a B lymphocyte stimulator, a member of the TNF ligand superfamily that exists in two forms, membrane-bound and soluble, and is specifically expressed on the surface of myeloid cells. It selectively stimulates B lymphocyte proliferation and immunoglobulin production. The term "APRIL" (a proliferation-inducing ligand) as used herein refers to a tumor necrosis factor (TNF) analog that can stimulate the proliferation of primitive B cells and T cells in the body and promote the accumulation of B cells. APRIL can specifically bind to TACI and BCMA, and after binding, can block the binding of APRIL to B cells and inhibit the APRIL-stimulated proliferation of primitive B cells. Furthermore, APRIL and BLys can competitively bind to receptors (BCMA, TACI).

[0030] As used herein, the term "TACI-Fc fusion protein" refers to a transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI)-immunoglobulin fusion protein (i.e., a TACI-Fc fusion protein). A TACI-immunoglobulin fusion protein according to the present invention comprises (i) the TACI extracellular domain or a fragment thereof that binds to Blys and / or APRIL, and (ii) a fragment of a human immunoglobulin constant domain.

[0031] For the term "TACI extracellular domain or a fragment thereof that binds to Blys and / or APRIL," see the TACI extracellular domain disclosed in U.S. Patent Nos. 5,969,102, 6,312,622, 6,500,428, and U.S. Patent Application Nos. 09 / 569,245 and 09 / 627,266 (the contents of which are incorporated herein by reference), as well as specific fragments of the TACI extracellular domain capable of interacting with a TACI ligand, or the amino acid fragment from positions 13 to 118 of the TACI extracellular domain disclosed in Chinese Patent Publication No. CN10,132,643A.

[0032] In the term "fragment of a human immunoglobulin constant region," the immunoglobulin is preferably IgG1, and may include a heavy chain constant region, such as a human heavy chain constant region. A preferred "fragment of a human immunoglobulin constant region" in the present invention is an amino acid fragment containing a portion of the hinge region domain, CH2 domain, and CH3 domain.

[0033] The term "treatment" as used herein, in relation to a given disease or condition, includes, but is not limited to, inhibiting the progression of the disease or condition, such as preventing the progression of the disease or condition, alleviating the disease or condition, such as eliminating the disease or condition, and diminishing the symptoms caused by the disease or condition, such as reducing or preventing or treating the symptoms of the disease or condition.

[0034] The term "telitacicept" (or "Tai'ai", which are used interchangeably in the present invention) in connection with the present invention is a type of TACI-Fc fusion protein, the amino acid sequence of which is shown in SEQ ID NO: 1 or as shown at https: / / extranet.who.int / soinn / mod / page / view.php?id=137&inn_n=10932. For its preparation method, see the disclosures in Chinese Patent Publication Nos. CN10,132,3643A or CN11,361,3675A. [Example]

[0035] Hereinafter, the present invention will be described in detail with reference to examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention.

[0036] [Example 1] Formulation Development In this example, several additives were screened and compared to study the effects of different concentrations of amino acids (e.g., arginine hydrochloride), surfactants (e.g., polysorbate 20 (Tween-20)), protectants (e.g., sucrose), and different pH values on the stability of TACI-Fc protein (having the amino acid sequence set forth in SEQ ID NO: 1). Histidine was used as a buffer (containing hydrochloric acid for pH adjustment), and an experimental design was performed with five factors and three levels using the CCF model in the software MODDE.

[0037] The TACI-Fc protein (SEQ ID NO: 1) solution was collected, concentrated by dialysis, added to the mother liquor, adjusted to a constant volume, filtered, and dispensed. The formulation information obtained is shown in Table 1 below. The prepared samples were stored at 25°C for 5 weeks (approximately 1 month longer), and SEC-purity was detected at 3 days (i.e., day 3, hereinafter the same), 7 days, 2 weeks (i.e., 2W, hereinafter the same), and 5 weeks. The 5-week study results were analyzed using the software MODDE.

[0038] [Table 1]

[0039] The experimental analysis results are shown in Figures 1, 2, 3, and 4. The results showed that the effects of arginine hydrochloride and sucrose concentrations on protein purity were statistically significant (P<0.05). With increasing arginine hydrochloride concentration, aggregates increased, resulting in a decrease in protein purity. On the other hand, with increasing sucrose concentration, aggregates decreased and protein purity increased. Surprisingly, the presence of Tween-20 had no effect on the reduction of aggregation and degradation, indicating that the addition of surfactants to the formulation is not necessary. The pH value did not have a significant effect on the change in monomer content (P>0.05), indicating that a pH range of 4.8 to 5.2 is suitable for telitacicept liquid protein formulations. The contour plot in Figure 5 indicates that the formulation without arginine hydrochloride, sucrose concentrations of 270 mmol / L to 300 mmol / L, and protein concentrations of 80 mg / mL are suitable.

[0040] [Example 2] Buffer system screening experiments In this example, to screen for the optimal buffer system for TACI-Fc protein formulations, the effects of three buffer systems, histidine-histidine hydrochloride, citrate, and succinate, were compared. The TACI-Fc protein (SEQ ID NO: 1) concentration was determined to be 160 mg / mL, pH 5.0, and sucrose concentration 290 mmol / L.

[0041] TACI-Fc protein (SEQ ID NO: 1) was collected and concentrated by dialysis according to the formulation in Table 2 below, then adjusted to a constant volume, filtered, and dispensed. The prepared samples were then stored at 25°C, and SEC-purity was measured at 3 and 9 days.

[0042] [Table 2]

[0043]

Table 3

[0044]

Table 4

[0045]

Table 5

[0046]

Table 6

[0047]

Table 7

[0048] The experimental results are shown in Tables 3 to 7. As a result, after standing at 25°C for 9 days, the purity of the main peak decreased by 1.2%, 2.6%, and 1.8%, respectively, in the histidine-histidine hydrochloride buffer system, the citrate buffer system, and the succinate buffer system. Furthermore, after standing at 25°C for 9 days, the amount of aggregates increased in all three buffer systems, and the amount of aggregates increased by 4.1%, 4.8%, and 3.7%, respectively, in the histidine-histidine hydrochloride buffer system, the citrate buffer system, and the succinate buffer system. In addition, the results of visible foreign matter and viscosity indicated that the citrate buffer system was less stable than the other two groups, exhibiting a cloudy appearance and high viscosity, making it unsuitable for protein preparation. Furthermore, dialysis of TACI-Fc protein with the citrate buffer system was more difficult than with the other two groups; the dialyzed protein formed a milky white suspension after standing at 4°C for a while and recovered to a clear white liquid after standing at room temperature for a while. Furthermore, the viscosity of the TACI-Fc protein after dialysis with the citrate buffer system was relatively higher than with the other two groups, and the protein solution was more viscous. These results suggest that the histidine-histidine hydrochloride and succinate buffer systems are superior to the citrate buffer system. Furthermore, at pH 5.0, the succinate buffer system demonstrated superior buffering properties to the histidine-histidine hydrochloride buffer system.

[0049] [Example 3] Protective agent screening experiment In Example 1, it was found that increasing the sucrose concentration improved protein stability. This is likely due to the fact that the hydroxyl groups in sucrose act as proton donors, inhibiting thiol deprotonation to some extent, thereby slowing the rate of disulfide-bonded aggregate formation. However, high concentrations of sucrose can significantly increase the osmotic pressure of liquid formulations, potentially affecting the isotonicity of the formulation required for subcutaneous injection and potentially leading to higher levels of endotoxin. After comparing and evaluating several different protective agents, this example focused on the effect of hydroxypropyl-β-cyclodextrin as a protective agent on formulation stability.

[0050] TACI-Fc protein (SEQ ID NO: 1) was collected and concentrated by dialysis according to the formulation in Table 8 below, adjusted to a constant volume, filtered, and dispensed. The prepared samples were then stored at 25°C and 40°C, respectively. SEC-purity was measured after 4 days and 7 days at 40°C, after 2 weeks, 3 weeks, and 4 weeks at 25°C, changes in capillary electrophoresis (nrCE-SDS) at 40°C, and the cellular activity and binding activity of the protein at 25°C.

[0051] [Table 8]

[0052] [Table 9]

[0053] [Table 10]

[0054] [Table 11]

[0055] [Table 12]

[0056] [Table 13]

[0057] The experimental results are shown in Tables 9 to 13. The SEC results at 40°C and 25°C (see Tables 9 to 10) indicated that the higher the hydroxypropyl-β-cyclodextrin concentration, the better the TACI-Fc protein stability; the higher the sucrose concentration, the better the protein stability; and the protective effect of hydroxypropyl-β-cyclodextrin on TACI-Fc protein was superior to that of sucrose. The nrCE-SDS results (see Table 11) indicated that the higher the hydroxypropyl-β-cyclodextrin concentration, the better the protein stability; the higher the sucrose concentration, the better the protein stability; and the protective effect of hydroxypropyl-β-cyclodextrin on protein was superior to that of sucrose, which is consistent with the results in Tables 9 and 10. The results of cell activity at 25°C (see Table 12) indicated no tendency for a decrease in cell activity in either case. The binding activity at 25°C (see 13 above) showed no tendency for a decrease in binding activity between the two cases. These results indicate that the presence of sucrose or hydroxypropyl-β-cyclodextrin reduces the occurrence of protein aggregation and degradation, and that hydroxypropyl-β-cyclodextrin provides better protection for the ACI-Fc protein than sucrose.

[0058] [Example 4] pH determination experiment In this example, we focused on the effect of different pH levels on the stability of TACI-Fc protein when the concentration of hydroxypropyl-β-cyclodextrin was 140 mmol / L and the concentration of the succinate buffer system was 10 mmol / L.

[0059] TACI-Fc protein (SEQ ID NO: 1) was dialyzed against 10 mmol / L succinic acid and concentrated to a protein concentration of 40 mg / mL, after which the volume was adjusted to a constant value. The mother liquor was added and then concentrated to the target concentration. The final formulation information is shown in Table 14 below. The prepared sample was then stored at 40°C, and the SEC purity was measured after 2 and 7 days.

[0060] [Table 14]

[0061] [Table 15]

[0062] The experimental results are shown in Table 15 above. As a result, it was observed that as the pH value increased, the amount of aggregates increased and decomposition decreased, but the increase in aggregates was greater than the decrease in decomposition, which indicated a decrease in monomers. In order to balance the degree of aggregates and decomposition, the pH was set to 5.0 to 5.2.

[0063] [Example 5] Hydroxypropyl-β-cyclodextrin concentration determination experiment In this example, we focused on the effects of 50 mmol / L, 100 mmol / L, and 140 mmol / L hydroxypropyl-β-cyclodextrin on protein stability when the pH was 5.1, the succinic acid concentration was 10 mmol / L, and the TACI-Fc (SEQ ID NO: 1) concentration was 80 mmol / L.

[0064] TACI-Fc protein (SEQ ID NO: 1) was concentrated by dialysis to the formulation shown in Table 16, adjusted to a constant volume, filtered, and dispensed. The prepared samples were then stored at 40°C, and the SEC purity, nrCE-SDS results, and osmolality were measured at 3 and 7 days.

[0065] [Table 16]

[0066] [Table 17]

[0067] [Table 18]

[0068] [Table 19]

[0069] The experimental results are shown in Tables 17 to 19. The results in Tables 17 and 18 indicate that the purity gradually decreases as the hydroxypropyl-β-cyclodextrin concentration decreases. The results in Table 19 indicate that an isotonic solution is obtained only when the hydroxypropyl-β-cyclodextrin concentration is 140 mmol / L. Therefore, in order to obtain a more appropriate solution osmotic pressure, the hydroxypropyl-β-cyclodextrin concentration must be increased in subsequent steps. The hydroxypropyl-β-cyclodextrin concentration in subsequent steps was set to 150 mmol / L, and this was determined through an osmotic pressure experiment.

[0070] [Experimental Example 6] Osmotic Pressure Determination Experiment Two parallel groups, A and B, were conducted as shown below. Specific prescription information is shown in Table 20 below.

[0071] [Table 20]

[0072] [Table 21]

[0073] The experimental results are shown in Table 21. From the results, when the concentration of TACI-Fc (SEQ ID NO: 1) was 80 mg / mL, the osmolality of both Groups A and B was approximately 330. When the concentration of TACI-Fc (SEQ ID NO: 1) was 120 mg / mL, the osmolality of both Groups A and B was approximately 400. The osmolality measured before and after filtration was not significantly different, indicating that filtration did not affect osmolality, and the pH was within the range of 5.1 ± 0.1. Based on the above, the composition of this formulation was determined to be 10 mmol / L succinic acid, 150 mmol / L hydroxypropyl-β-cyclodextrin, a protein concentration of 80 mg / mL, and a pH of 5.1 ± 0.1 adjusted with sodium hydroxide.

[0074] [Example 7] Long-term stability experiments In this example, the long-term stability and accelerated stability of the final formulation concentrates after packaging in the respective precharged syringes were investigated.

[0075] TACI-Fc protein (SEQ ID NO: 1) was concentrated by dialysis to the following formulations, adjusted to a constant volume, filtered, and dispensed. Specific formulation information is shown in Table 22 below. The long-term stability of the samples at different temperatures was investigated according to the temperature and time parameters in Table 23 below.

[0076] [Table 22]

[0077] [Table 23]

[0078] SEC-purity data showed that after 24 months of storage of TACI-Fc (SEQ ID NO: 1) at -20 to 4°C, aggregates increased from 1.3% to 2.88%, and purity decreased from 98.6% to 94.7%, but both were within the quality standard range. Furthermore, no increase in particulate matter (visible foreign matter and insoluble particles) was observed, and all results were within the acceptable standard range. Other product quality attribute parameters, including peptide map, appearance, osmolality, pH, CE-SDS (r / nr), RP-HPLC, activity, protein content, bacterial endotoxin, and container closure integrity (CCI), remained largely unchanged, and all results were within the acceptable standard range. Rigorous long-term stability evaluation tests revealed that the formulation of the present invention, when filled into a prefilled syringe (PFS) and stored for long periods at -20 to 4°C, exhibited good stability and met all requirements of the stability and quality standards. In addition, SEC-purity measurement experiments revealed that even after 6 months of storage at 25°C, there was a slight increase in aggregates, but all detection results were within the quality standard range.

[0079] The gist of the present invention has been described in detail through the preferred embodiments of the present invention. Those skilled in the art will understand that any modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention are included within the protection scope of the present invention.

Claims

1. 1. A liquid formulation of a transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI)-Fc fusion protein, comprising a TACI-Fc fusion protein, a protectant, a buffer, and a pH adjuster, the TACI-Fc fusion protein comprises (i) the extracellular domain of TACI or a fragment thereof that binds to B-lymphocyte stimulatory factor (Blys) and / or proliferation-inducing ligand (APRIL); and (ii) a fragment of a human immunoglobulin constant domain; the protecting agent is hydroxypropyl-β-cyclodextrin; the buffer is succinic acid; A liquid formulation of a TACI-Fc fusion protein, wherein the pH of the liquid formulation is in the range of 5.0 to 5.

2.

2. 2. The liquid formulation of claim 1, wherein the concentration of the TACI-Fc fusion protein ranges from 40 mg / ml to 240 mg / ml.

3. 4. The liquid formulation of claim 3, wherein the concentration of the TACI-Fc fusion protein is about 40 mg / ml, 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml, 150 mg / ml, 160 mg / ml, 170 mg / ml, 180 mg / ml, 190 mg / ml, 200 mg / ml, 210 mg / ml, 220 mg / mL, 230 mg / ml, or 240 mg / ml.

4. The liquid formulation of TACI-Fc fusion protein according to claim 2 or 3, characterized in that the concentration of the protecting agent is in the range of 100 mmol / L to 200 mmol / L.

5. The liquid formulation of TACI-Fc fusion protein of claim 4, wherein the concentration of the protecting agent is about 100 mmol / L, 110 mmol / L, 120 mmol / L, 130 mmol / L, 140 mmol / L, 150 mmol / L, 160 mmol / L, 170 mmol / L, 180 mmol / L, 190 mmol / L, or 200 mmol / L.

6. The liquid formulation of TACI-Fc fusion protein according to claim 4 or 5, wherein the concentration of the buffer is in the range of 5 mmol / L to 15 mmol / L.

7. The liquid formulation of TACI-Fc fusion protein of claim 6, wherein the concentration of the buffer is about 5 mmol / L, 6 mmol / L, 7 mmol / L, 8 mmol / L, 9 mmol / L, 10 mmol / L, 11 mmol / L, 12 mmol / L, 13 mmol / L, 14 mmol / L, or 15 mmol / L.

8. The liquid formulation of TACI-Fc fusion protein according to claim 6 or 7, wherein the pH adjusting agent is an alkaline inorganic salt.

9. The liquid formulation of TACI-Fc fusion protein of claim 8, wherein the pH adjusting agent is sodium hydroxide.

10. The liquid formulation of TACI-Fc fusion protein according to claim 8 or 9, wherein the pH of the liquid formulation is about 5.0, 5.1, or 5.

2.

11. The liquid formulation of TACI-Fc fusion protein according to claim 10, wherein the TACI-Fc fusion protein has the amino acid sequence set forth in SEQ ID NO:

1.

12. The liquid formulation of TACI-Fc fusion protein of claim 11, wherein the TACI-Fc fusion protein is telitacicept.

13. A liquid formulation of a TACI-Fc fusion protein according to claim 11 or 12, characterized in that the liquid formulation comprises approximately 10 mmol / L succinic acid, approximately 150 mmol / L hydroxypropyl-β-cyclodextrin, approximately 80 mg / mL of the TACI-Fc fusion protein, and the pH of the liquid formulation is approximately 5.

1.

14. 13. The liquid formulation of claim 11 or 12, wherein the liquid formulation contains approximately 80 mg of TACI-Fc fusion protein, approximately 1.18 mg of succinic acid, and approximately 209.85 mg of hydroxypropyl-β-cyclodextrin per ml, and the pH of the liquid formulation is approximately 5.

1.

15. Use of a liquid formulation of a TACI-Fc fusion protein according to any one of claims 1 to 14 in the manufacture of a medicament for preventing, alleviating and / or treating an autoimmune disease.

16. A liquid formulation of the TACI-Fc fusion protein of any one of claims 1 to 14 for preventing, alleviating and / or treating autoimmune diseases.

17. A method for preventing, alleviating and / or treating an autoimmune disease, comprising administering to a subject in need thereof the liquid formulation of any one of claims 1 to 14.

18. A packaged pharmaceutical product comprising a liquid formulation of a TACI-Fc fusion protein according to any one of claims 1 to 14.

19. A pre-filled syringe containing a liquid formulation of the TACI-Fc fusion protein of any one of claims 1 to 14.

20. 20. The syringe of claim 19, wherein the syringe is an auto-injector pre-filled syringe.

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