Pharmaceutical preparation for subcutaneous administration, comprising antibody-drug conjugate and hyaluronidase

Abstract

The present invention relates to a pharmaceutical preparation for subcutaneous administration, the preparation comprising: an antibody-drug conjugate (ADC); and native human recombinant hyaluronidase PH20.

Description

Pharmaceutical preparations for subcutaneous administration comprising antibody-drug conjugates and hyaluronidase

[0001] The present invention relates to a pharmaceutical formulation for subcutaneous administration comprising a highly concentrated antibody-drug conjugate and hyaluronidase.

[0002] Unlike conventional antibodies, antibody-drug conjugates (ADCs) are a form in which an antibody and a drug are combined through a linker. In antibody-drug conjugates, the drug is not used alone because its strong toxicity can cause toxicity even to normal cells; instead, it is characterized by being selectively delivered through the antibody.

[0003] Various prior studies have been conducted on administering antibody-drug conjugates via intravenous injection. However, this intravenous method is costly, cumbersome, and causes patient discomfort, aversion, and side effects due to frequent administration, and it is limited to medically trained individuals. In particular, since the maximum plasma concentration is reached immediately after pharmacokinetic intravenous administration, increasing the dose of the antibody-drug conjugate increases the probability of systemic toxic reactions due to the toxicity of the drug within the conjugate.

[0004] There is a need for research that can effectively diffuse antibody-drug conjugates while resolving the issues associated with intravenous administration. Accordingly, if the intravenous method is changed to subcutaneous administration, systemic toxic reactions are expected to be significantly reduced because, pharmacokinetically, plasma concentrations increase slowly after subcutaneous administration and the maximum plasma concentration is relatively low. However, in concentrating antibody-drug conjugates to high concentrations, it is necessary to improve the solubility of the antibody-drug conjugate through formulation research.

[0005] The present invention aims to provide a pharmaceutical formulation for subcutaneous administration comprising an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20.

[0006] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below.

[0007] The present invention provides a pharmaceutical formulation for subcutaneous administration comprising an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20.

[0008] In the above antibody-drug conjugate, the antibody may comprise one or more selected from the group consisting of trastuzumab, sacituzumab, datopotamab, gemtuzumab, inotuzumab, moxetumomab, brentuximab, poletuzumab, enfortumab, belantamab, loncastuximab, mirvetuximab, and tositumomab.

[0009] In the above antibody-drug conjugate, the drug may have a molecular weight of 100 g / mol to 5,000 g / mol.

[0010] The above antibody-drug conjugates are Trastuzumab-deruxtecan, Trastuzumab-emtansine, Sacituzumab-govitecan, Datopotamab-deruxtecan, Gemtuzumab-ozogamicin, Inotuzumab-ozogamicin, Moxetumomab-pasudotox, Brentuximab-vedotin, Polatuzumab-vedotin, Enfortumab-vedotin, and Belantab-mafodotin, It may include one or more selected from the group consisting of Loncastuximab-tesirine and Mirvetuximab-soravtansine.

[0011] The above antibody-drug conjugate may be selected from the group consisting of trastuzumab-deruxtecan, trastuzumab-emtansine, and sacituzumab-govitecan.

[0012] The content of the above antibody-drug conjugate may be 20 mg / mL to 240 mg / mL.

[0013] The content of the above antibody-drug conjugate may be 50 mg / mL to 240 mg / mL.

[0014] The above natural human recombinant hyaluronidase PH20 may be for the diffusion of the above antibody-drug conjugate.

[0015] The content of the above natural human recombinant hyaluronidase PH20 may be 1,000 unit / mL to 10,000 unit / mL.

[0016] When the above pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the area under the concentration-time curve (AUC) of the antibody in plasma may be 105% or more compared to when a preparation containing the antibody-drug conjugate alone is administered subcutaneously at the same dose.

[0017] As a buffer, it may additionally contain succinic acid or a salt thereof; or sodium phosphate.

[0018] As a buffer, it may further comprise 1 mM to 20 mM of succinic acid or a salt thereof; or sodium phosphate.

[0019] As a surfactant, one or more selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80; as a sugar or derivative thereof, one or more selected from the group consisting of sorbitol, mannitol, trehalose and sucrose; and as a viscosity reducing agent, methionine or arginine may be further included.

[0020] It may further comprise 0.05 mg / mL to 5 mg / mL of polysorbate 80 as a surfactant; 10 mg / mL to 100 mg / mL of sorbitol as a sugar or derivative thereof; and 0.5 mg / mL to 20 mg / mL of methionine as a viscosity reducing agent.

[0021] The pharmaceutical formulation for subcutaneous administration according to the present invention comprises an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20. By combining the antibody-drug conjugate with natural human recombinant hyaluronidase PH20, the antibody-drug conjugate can be effectively diffused. In particular, when the content of the antibody-drug conjugate is concentrated to a high concentration, the addition of natural human recombinant hyaluronidase PH20 has the advantage of maximizing the diffusion effect of the antibody-drug conjugate while reducing local toxicity reactions. Furthermore, as subcutaneous administration gradually increases the plasma concentration pharmacokinetically and decreases the maximum plasma concentration, systemic toxicity reactions can also be significantly reduced.

[0022] Furthermore, by optimizing the formulation of the pharmaceutical preparation for subcutaneous administration according to the present invention, stability can be ensured while maintaining a viscosity suitable for subcutaneous administration.

[0023] In particular, the pharmaceutical formulation for subcutaneous administration according to the present invention can significantly improve the transparency and stability of the antibody-drug conjugate provided for subcutaneous administration by using succinic acid, its salt, or sodium phosphate as a buffer. Furthermore, it has the advantage of not causing significant changes based on the target pH for each antibody-drug conjugate and maintaining the natural human recombinant hyaluronidase pH 20 potency at a high level.

[0024] Figure 1 is a graph showing the results of analyzing changes in plasma trastuzumab concentration over time after administering the drugs of Example 1 and Comparative Example 1 to rats once via subcutaneous injection and collecting blood up to 28 days.

[0025] Figure 2 is a graph showing the results of analyzing the change in plasma concentration of sasituzumab over time after administering the drugs of Example 2 and Comparative Example 2 to rats once via subcutaneous injection and collecting blood up to 28 days.

[0026] Figures 3(a)-(b) are graphs showing the results of analyzing changes in plasma trastuzumab concentration over time after administering the subcutaneous formulations of Examples 3-1 and 4-1 and Comparative Examples 3-1 and 4-1 to rats once via subcutaneous injection and collecting blood up to 28 days.

[0027] Figures 4(a)-(b) are graphs showing the results of analyzing changes in plasma concentration of sasituzumab over time after administering the subcutaneous formulations of Examples 5-1 and 6-1 and Comparative Examples 5-1 and 6-1 to rats once via subcutaneous injection and collecting blood up to 28 days.

[0028] The inventors confirmed that in preparing an antibody-drug conjugate as a pharmaceutical formulation for subcutaneous administration, the antibody-drug conjugate can be effectively diffused by using a natural human recombinant hyaluronidase PH20 in combination with a highly concentrated antibody-drug conjugate, and derived an optimal formulation to ensure stability under refrigerated conditions. Thus, the present invention was completed.

[0029]

[0030] The present invention will be described in detail below.

[0031]

[0032] Pharmaceutical preparations for subcutaneous administration comprising antibody-drug conjugates and hyaluronidase

[0033]

[0034] The present invention provides a pharmaceutical formulation for subcutaneous administration comprising an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20.

[0035]

[0036] First, the pharmaceutical formulation for subcutaneous administration according to the present invention comprises an antibody-drug conjugate (ADC).

[0037]

[0038] The above antibody-drug conjugate may consist of an antibody, a drug, and a linker for combining the antibody and the drug.

[0039]

[0040] Specifically, in the above antibody-drug conjugate, the antibody not only possesses targetability, efficiency, and stability by targeting a specific antigen, but can also function as a therapeutic antibody as needed. Specifically, the antibodies are trastuzumab targeting HER2, sacituzumab or datopotamab targeting Trop-2, gemtuzumab targeting CD33, innotuzumab or moxetumomab targeting CD22, brentuximab targeting CD30, polatuzumab targeting CD79B, enfortumab targeting Nectin-4, belantamab targeting BCMA (B-cell maturation antigen), loncastuximab targeting CD19, mirvetuximab targeting FRα (Folate receptor α), and targeting CD20 It may include one or more selected from the group consisting of tositumomab, and preferably trastuzumab, which is approved as a treatment for HER2-positive early / metastatic breast cancer, HER2-positive advanced / metastatic gastric cancer or HER2-positive gastroesophageal junction adenocarcinoma, or sacituzumab, which is approved as a treatment for triple-negative breast cancer, HR+ / HER2- advanced / metastatic breast cancer or metastatic urothelial carcinoma, but is not limited thereto.

[0041]

[0042] In the above antibody-drug conjugate, the drug is capable of being bound to the antibody through a linker, and the drug preferably has a molecular weight of 100 g / mol to 5,000 g / mol, but is not limited thereto.

[0043] Specifically, the drug may comprise one or more selected from the group consisting of deruxtecan derivative, emtansine derivative, govitecan derivative, ozogamicin derivative, pasudotox derivative, vedotin derivative, mafodotin derivative, tesirine derivative, and soravtansine derivative, and the drug may be a topoisomerase I inhibitor, specifically a deruxtecan derivative (DX-8951 derivative (DXd)) (molecular weight: about 796.88 g / mol) or a govitecan derivative (molecular weight: about 392.40 g / mol). These are characterized by having strong toxicity and being selectively delivered to bind to a specific antibody to inhibit topoisomerase I. Alternatively, the drug may be an emtansine derivative (molecular weight: approximately 622.86 g / mol) as a microtubule formation inhibitor. This is also characterized by having strong toxicity and being selectively delivered to bind to a specific antibody to inhibit microtubule formation.

[0044]

[0045] In the above antibody-drug conjugate, the linker is for binding the antibody and the drug. Specifically, the linker may be a known linker, may be a hydrolyzable linker or a non-hydrolyzable linker, and is preferably a hydrolyzable linker (particularly a tetrapeptide linker), but is not limited thereto. For example, one end of the linker may be bound to the end of a deruxtecan derivative, which is a type of drug, and in this case, the total molecular weight of the linker and the drug (i.e., deruxtecan) may be about 1034.05 g / mol.

[0046]

[0047] More specifically, the antibody-drug conjugate is trastuzumab-deruxtecan, trastuzumab-emtansine, sacituzumab-govitecan, datopotamab-deruxtecan, gemtuzumab-ozogamicin, inotuzumab-ozogamicin, moxetumomab-pasudotox, brentuximab-vedotin, polatuzumab-vedotin, enfortumab-vedotin, belantamab-mafodotin, It may include one or more selected from the group consisting of Loncastuximab-tesirine and Mirvetuximab-soravtansine, and preferably, but not limited to, one or more selected from the group consisting of Trastuzumab-deruxtecan, Trastuzumab-emtansine and Sacituzumab-govitecan.

[0048]

[0049] Meanwhile, the antibody-drug conjugate is characterized by being concentrated to a high concentration, wherein the content of the antibody-drug conjugate may be 20 mg / mL to 240 mg / mL, preferably 50 mg / mL to 240 mg / mL, more preferably 60 mg / mL to 240 mg / mL, and more preferably 90 mg / mL to 240 mg / mL, but is not limited thereto. At this time, if the content of the antibody-drug conjugate is too low, there is a problem that the expression of an effective drug effect is insufficient, and if the content of the antibody-drug conjugate is too high, it may be difficult to implement it as a subcutaneous administration formulation.

[0050] In particular, even within the content range under high concentration conditions as described above, the synergistic effect resulting from the addition of the natural human recombinant hyaluronidase PH20 described later may become more pronounced as the content of the antibody-drug conjugate increases.

[0051] Meanwhile, when the above-mentioned pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the single dose of the antibody-drug conjugate may be 10 mg to 6,000 mg, preferably 25 mg to 6,000 mg, more preferably 30 mg to 6,000 mg, more preferably 45 mg to 6,000 mg, and most preferably 45 mg to 1,800 mg, but is not limited thereto. To increase the single dose of the antibody-drug conjugate, the content of the antibody-drug conjugate itself may be increased, but the administration volume may be increased while maintaining the content to implement it as a subcutaneous administration formulation. In this case, the administration volume may be 0.5 mL to 25 mL.

[0052]

[0053] As mentioned above, the antibody-drug conjugate requires effective diffusion. In the case of the antibody-drug conjugate, in particular, it is necessary to select and use natural human recombinant hyaluronidase PH20 as a diffusion agent to optimize its diffusion by comprehensively considering the mechanism of action, structure, and molecular weight of the drug. Through this combination, there is an advantage of effectively diffusion of the antibody-drug conjugate while reducing local / systemic toxicity.

[0054] Accordingly, the pharmaceutical formulation for subcutaneous administration according to the present invention comprises natural human recombinant hyaluronidase PH20, wherein the natural human recombinant hyaluronidase PH20 is intended to effectively diffuse the antibody-drug conjugate while reducing local toxicity.

[0055] The above natural human recombinant hyaluronidase PH20 may use human-derived hyaluronidase PH20 known in the art, and is preferably represented by the amino acid sequence of SEQ ID NO. 1, but is not limited thereto.

[0056] Unlike its PH20 variant, the above-mentioned natural human recombinant hyaluronidase PH20 is an endogenous protein (enzyme) that has the same amino acid sequence and three-dimensional structure as the PH20 present in humans. Accordingly, it has the advantage of being safe when administered to the human body, as it is less likely to act as an antigen when injected externally.

[0057] The content of the above-mentioned natural human recombinant hyaluronidase PH20 may be 1,000 unit / mL to 10,000 unit / mL, preferably 1,000 unit / mL to 5,000 unit / mL, and more preferably 1,000 unit / mL to 4,000 unit / mL, but is not limited thereto. At this time, if the content of the above-mentioned natural human recombinant hyaluronidase PH20 is too low, there is a problem that the diffusion effect of the antibody-drug conjugate is insufficient, and if the content of the above-mentioned natural human recombinant hyaluronidase PH20 is too high, it may be difficult to implement it as a subcutaneous administration formulation.

[0058] Meanwhile, when the above-mentioned pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the single dose of recombinant hyaluronidase PH20 may be 500 units to 250,000 units, preferably 500 units to 125,000 units, more preferably 500 units to 100,000 units, and most preferably 500 units to 30,000 units, but is not limited thereto. In order to increase the single dose of the above-mentioned natural human recombinant hyaluronidase PH20, the content of the above-mentioned natural human recombinant hyaluronidase PH20 itself may be increased, but the administration volume may also be increased while maintaining the content to implement it as a subcutaneous administration formulation. In this case, the administration volume may be 0.5 mL to 25 mL.

[0059]

[0060] Thus, when the above-mentioned pharmaceutical preparation for subcutaneous administration is administered subcutaneously, compared to the case where a preparation containing an antibody-drug conjugate alone is administered subcutaneously at the same dose, the area under the concentration-time curve (AUC) of the antibody in plasma may be 105% or more, and preferably 115% or more, but is not limited thereto. Meanwhile, the maximum plasma concentration of the antibody (C max The value can be 120% or more.

[0061] The above drug transfer amount (AUC last and C max Through the increase of ), the effective diffusion of the antibody-drug conjugate following the addition of the natural human recombinant hyaluronidase PH20 can be confirmed. In particular, regarding the antibody-drug conjugate, even within the content range under high concentration conditions as described above, it can be confirmed that the synergistic effect following the addition of the natural human recombinant hyaluronidase PH20 becomes more pronounced as the content of the antibody-drug conjugate increases.

[0062]

[0063] Furthermore, by optimizing the formulation of the pharmaceutical preparation for subcutaneous administration according to the present invention, stability can be ensured while maintaining a viscosity suitable for subcutaneous administration.

[0064] In the pharmaceutical formulation for subcutaneous administration according to the present invention, the viscosity under a temperature condition of 25°C may be 1 mPa / s to 25 mPa / s, preferably 1 mPa / s to 12 mPa / s, and more preferably 1 mPa / s to 6 mPa / s (e.g., 2 mPa / s to 6 mPa / s or 3 mPa / s to 5 mPa / s), but is not limited thereto. That is, by maintaining the above viscosity range, a gelatinous appearance may not be observed at all, and when used as a pharmaceutical formulation for subcutaneous administration, side effects such as the possibility of tissue damage at the injection site, pain, and improper distribution of the drug may not occur.

[0065] In this specification, "viscosity under 25°C temperature conditions" refers to the dynamic viscosity representing internal frictional resistance when a pharmaceutical formulation for subcutaneous administration flows, which may be measured immediately after manufacturing or after storage under refrigerated conditions (2-8°C) for up to 8 weeks (preferably within 4 weeks). In this case, a commercial viscometer such as m-VROC may be used to measure the viscosity.

[0066] In addition, the pharmaceutical formulation for subcutaneous administration according to the present invention is characterized by ensuring stability (particularly, stability under refrigerated conditions), and is characterized by maintaining the content, purity, and drug-antibody ratio (DAR) of the antibody-drug conjugate constant after refrigerated storage.

[0067]

[0068] Meanwhile, the above antibody-drug conjugate may be provided in powder (or freeze-dried) and liquid forms, and stability issues such as visible particles (precipitation) and layer separation may occur when storing the antibody-drug conjugate.

[0069] To solve these problems, the pharmaceutical formulation for subcutaneous administration according to the present invention may additionally include succinic acid or a salt thereof; or sodium phosphate as a buffer. In this case, succinic acid or a salt thereof as the buffer may refer to a succinate buffer in which sodium hydroxide (NaOH) is mixed with succinic acid. Such an optimal buffer can significantly improve the transparency and stability of the antibody-drug conjugate provided for subcutaneous administration. Furthermore, it has the advantage of not causing significant changes based on the target pH for each antibody-drug conjugate and maintaining the PH20 titer of the natural human recombinant hyaluronidase at a high level.

[0070] Specifically, the above buffer solution can store a highly concentrated antibody-drug conjugate under stable conditions by maintaining the pH of the pharmaceutical preparation for subcutaneous administration at 4.5 to 7.0. In particular, since the target pH may differ depending on the antibody-drug conjugate, it is necessary to maintain the pharmaceutical preparation for subcutaneous administration at pH 5.0 to 6.0 when the antibody-drug conjugate is trastuzumab-deruxtecan, at pH 4.5 to 5.5 when the antibody-drug conjugate is trastuzumab-emtansine, and at pH 6.0 to 7.0 when the antibody-drug conjugate is sacituzumab-govitecan.

[0071] More specifically, the buffer may be succinic acid or a salt thereof or sodium phosphate, and its concentration may be 1 mM to 20 mM, preferably 5 mM to 15 mM, but is not limited thereto.

[0072] Meanwhile, when sodium citrate buffer is applied, there may be limitations in terms of stability, as a very large number of visible particles or separation of layers is observed in some antibody-drug conjugate formulations.

[0073]

[0074] In addition, when MES buffer is applied, it was confirmed that the pH change relative to the target pH after refrigeration is relatively large, so it may not be appropriate in terms of stability, and in particular, there may be a limitation in that it significantly reduces the activity of natural human recombinant hyaluronidase pH 20 in the trastuzumab-deruxtecan formulation after 4 weeks of refrigeration.

[0075]

[0076] Optionally, the pharmaceutical formulation for subcutaneous administration according to the present invention may further comprise, as a surfactant, one or more selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and poloxamer 188; as a sugar or derivative thereof, one or more selected from the group consisting of sorbitol, mannitol, trehalose, and sucrose; and as a viscosity reducing agent, one or more selected from the group consisting of methionine, arginine, proline, glycine, and histidine.

[0077] Specifically, the surfactant is intended to prevent aggregation of a highly concentrated antibody-drug conjugate and may be one or more selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and poloxamer 188, preferably polysorbate 80 with an amount of 0.05 mg / mL to 5 mg / mL, more preferably polysorbate 80 with an amount of 0.1 mg / mL to 1 mg / mL, but is not limited thereto. In addition, the above sugar or its derivative acts as a protein stabilizer and is intended to suppress osmotic stress and reduce viscosity. It may be one or more selected from the group consisting of sorbitol, mannitol, trehalose, and sucrose, preferably sorbitol in an amount of 10 mg / mL to 100 mg / mL, and more preferably sorbitol in an amount of 30 mg / mL to 50 mg / mL, but is not limited thereto. In addition, the above viscosity reducing agent is also intended to reduce viscosity and may be one or more selected from the group consisting of methionine, arginine, proline, glycine, and histidine. It is preferably methionine in an amount of 0.5 mg / mL to 20 mg / mL, and more preferably methionine in an amount of 0.5 mg / mL to 5 mg / mL, but is not limited thereto.

[0078]

[0079] Thus, the pharmaceutical formulation for subcutaneous administration according to the present invention has an anticancer effect and can target HER2, Trop-2, CD33, CD22, CD30, CD79B, Nectin 4, BCMA (B-cell maturation antigen), CD19, FRα (Folate receptor α), or CD20, and it is more preferable to target HER2 or Trop-2, but is not limited thereto. Specifically, the pharmaceutical formulation for subcutaneous administration according to the present invention may have a preventive or therapeutic effect against one or more cancers, including breast cancer, gastric cancer, adenocarcinoma, urothelial carcinoma, ovarian cancer, lung cancer, lymphoma, pancreatic cancer, etc.

[0080] In addition, the pharmaceutical formulation for subcutaneous administration according to the present invention is limited to a subcutaneous injection method in which the administration is carried out into the adipose tissue of the subcutaneous tissue located below the epidermis and dermis, and the administration cycle can be controlled by the specific type and dose of the antibody-drug conjugate and the diffusion action of natural human recombinant hyaluronidase PH20.

[0081]

[0082] Alternatively, the present invention provides a method for subcutaneously administering to an individual a pharmaceutical preparation comprising an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20.

[0083] Alternatively, the present invention provides a method for preventing or treating cancer comprising the step of subcutaneously administering to an individual a pharmaceutical preparation comprising an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20. In this case, "individual" refers to a subject requiring treatment for a disease, and more specifically, to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle.

[0084]

[0085] As reviewed above, the pharmaceutical formulation for subcutaneous administration according to the present invention comprises an antibody-drug conjugate (ADC); and natural human recombinant hyaluronidase PH20. By combining the antibody-drug conjugate with natural human recombinant hyaluronidase PH20, there is an advantage of effectively diffusing the antibody-drug conjugate. In particular, when the content of the antibody-drug conjugate is concentrated to a high concentration, the addition of natural human recombinant hyaluronidase PH20 has the advantage of maximizing the diffusion effect of the antibody-drug conjugate while reducing local toxicity reactions. Furthermore, as subcutaneous administration gradually increases the plasma concentration pharmacokinetically and decreases the maximum plasma concentration, systemic toxicity reactions can also be significantly reduced.

[0086] Furthermore, by optimizing the formulation of the pharmaceutical preparation for subcutaneous administration according to the present invention, stability can be ensured while maintaining a viscosity suitable for subcutaneous administration.

[0087] In particular, the pharmaceutical formulation for subcutaneous administration according to the present invention can significantly improve the transparency and stability of the antibody-drug conjugate provided for subcutaneous administration by using succinic acid, its salt, or sodium phosphate as a buffer. Furthermore, it has the advantage of not causing significant changes based on the target pH for each antibody-drug conjugate and maintaining the natural human recombinant hyaluronidase pH 20 potency at a high level.

[0088]

[0089] Preferred embodiments are presented below to aid in understanding the present invention. However, the following embodiments are provided merely to facilitate a better understanding of the invention, and the scope of the invention is not limited by the following embodiments.

[0090]

[0091] [Example]

[0092] Example 1 and Comparative Example 1: Trastuzumab diffusion effect depending on the addition of natural human recombinant hyaluronidase pH 20

[0093] Natural Human Recombinant Hyaluronidase PH20 (rHuPH20, Product Name: HyDIFFUZE TM To confirm the diffusion effect of trastuzumab due to the addition of ) a drug containing only trastuzumab-deruxtecan 40 mg / mL was designated as Comparative Example 1, wherein HyDIFFUZE was added to trastuzumab-deruxtecan 40 mg / mL TM Drugs with 2,000 unit / mL added were prepared as Example 1. Referring to Table 1, each drug was administered to rats once via subcutaneous injection, and plasma was collected up to 28 days.

[0094] Classification Trastuzumab-Deruxtecan Content / Single Dose HyDIFFUZE TM Content / Single Dose Comparison Example 1 Trastuzumab-deruxtecan 40 mg / mL - Trastuzumab-deruxtecan 84 mg / kg - Example 1 Trastuzumab-deruxtecan 40 mg / mL HyDIFFUZE TM 2,000 units / mLTrastuzumab-deruxtecan 84 mg / kgHyDIFFUZE TM 4,200 unit / kg

[0095] Collected plasma samples were analyzed using a Trastuzumab ELISA Kit (Matriks biotek, Cat. No. TRA-FD-HH), and pharmacokinetic analysis was performed using the WinNonlin program based on the analysis results (Table 2 and Figure 1).

[0096] Classification AUC last (ug·hr / mL)C max (ug / mL)T max (hr)t 1 / 2(hr)CL t (mL / hr / kg)Vd(mL / kg)MRT(hr) Comparative Example 1 Average 140 488.40 354.8 212 0.00 109.9 20.58 91.6 252.13 Standard Deviation 40 24.56 17.22 44.90 10.60 0.0 26.8 9.29 N5555555 Example 1 Average 149 944.40 448.6 67.20 110.6 20.55 87.24 235.27 Standard Deviation 13 480.5 374.5 210.7 316.21 0.0 513.88 13.40 N5555555 Example 1's relative to Comparative Example 1 Percentage(%) 106.731 26.4556.001 00.6494.8395.1693.31

[0097] As a result, in the case of Example 1, where the trastuzumab-deruxtecan content was 40 mg / mL, compared to Comparative Example 1, which had an equivalent trastuzumab-deruxtecan content / dose, the drug transfer amount (AUC last and C max As it is confirmed that ) increases, it is judged that the drug diffusion effect is excellent. In addition, compared to Comparative Example 1, the drug transfer rate (1 / T max It is also confirmed that ) increases. Accordingly, it is believed that local toxicity can be reduced by decreasing the time the drug remains subcutaneously.

[0098] In other words, HyDIFFUZE to trastuzumab-deruxtecan TM It is confirmed that the addition of can increase the overall absorption kinetics of trastuzumab.

[0099]

[0100] Example 2 and Comparative Example 2: Sassituzumab Diffusion Effect Depending on the Addition of Natural Human Recombinant Hyaluronidase PH20

[0101] Natural Human Recombinant Hyaluronidase PH20 (rHuPH20, Product Name: HyDIFFUZE TMTo confirm the diffusion effect of sacituzumab following the addition of ) a drug containing only sacituzumab-govitecan 120 mg / mL was designated as Comparative Example 2, wherein HyDIFFUZE to sacituzumab-govitecan 120 mg / mL TM Drugs with 2,000 unit / mL added were prepared as Example 2. Referring to Table 3, each drug was administered to rats once via subcutaneous injection, and plasma was collected up to 28 days.

[0102] HyDIFFUZE sacituzumab-govitecan content / single dose TM Content / Single Dose Comparison Example 2 Sacituzumab-govitecan 120 mg / mL Sacituzumab-govitecan 84 mg / kg Example 2 Sacituzumab-govitecan 120 mg / mL HyDIFFUZE TM 2,000 units / mLSacituzumab-govitecan 84 mg / kgHyDIFFUZE TM 1,400 unit / kg

[0103] Collected plasma samples were analyzed using a Sacituzumab ELISA Kit (Abbexa, Cat. No abx395055), and pharmacokinetic analysis was performed using the WinNonlin program based on the analysis results (Table 4 and Figure 2).

[0104] Classification AUC last (ug·hr / mL)C max (ug / mL)T max (hr)t 1 / 2 (hr)CL t(mL / hr / kg)Vd(mL / kg)MRT(hr) Comparative Example 2 Average 448 25.8 321 6.30 48.00 124.48 1.8 232 2.76 174.95 Standard Deviation 42 30.2 35 6.0 20.00 26.5 30.2 15 4.6 31 3.91 N4444444 Example 2 Average 527 59.9 435 3.30 24.00 115.1 51.56 25 8.6 21 46.03 Standard Deviation 40 25.1 37 6.8 00.00 7.7 10.1 22 1.20 12.19 N4444444 Example 2's relative to Comparative Example 2 Percentage(%)117.70163.3450.0092.5085.7180.1383.47

[0105] As a result, in the case of Example 2, in which the sacituzumab-govitecan content was highly concentrated at 120 mg / mL, the drug transfer amount (AUC) was compared to Comparative Example 2, which had an equivalent sacituzumab-govitecan content / dose. last and C max It is confirmed that ) is increasing further, and in particular, AUC last and C max It is confirmed that they increased significantly by approximately 117% and 163%, respectively. In addition, the drug transfer rate (1 / T max It is also confirmed that it increases even more significantly, by about twofold. Accordingly, it is believed that local toxicity can be reduced by shortening the time the drug remains subcutaneously.

[0106] That is, HyDIFFUZE in highly concentrated sacituzumab-govitecan (sacituzumab-govitecan content: ≥ 50 mg / mL) TM By adding it, the diffusion effect of the Sasituzumap can be maximized, so it can be considered effective.

[0107]

[0108] In summary, based on the findings in Tables 1 to 4, HyDIFFUZE at 20 mg / mL or higher of antibody-drug conjugate TMBy adding, a diffusion effect of antibodies may be achieved. In particular, when the antibody-drug conjugate is concentrated to a high concentration of 50 mg / mL or more, HyDIFFUZE TM The diffusion effect of the antibody-drug conjugate can be maximized through addition.

[0109]

[0110] Examples 3-5: Derivation of subcutaneous formulations to improve the stability of antibody-drug conjugates

[0111] A subcutaneous formulation was developed to improve the stability of the antibody-drug conjugate while having a viscosity desirable for subcutaneous administration.

[0112] Specifically, HyDIFFUZE in an antibody-drug conjugate 120 mg / mL TM Pharmaceutical formulations for subcutaneous administration were prepared in Examples 3 to 5, each comprising 2,000 unit / mL of a total of four types of buffers [succinate, sodium phosphate, MES, and sodium citrate], polysorbate 80 as a surfactant, sorbitol as a sugar or derivative thereof, and methionine as a viscosity reducing agent (Table 5).

[0113] HyDIFFUZE Antibody-Drug Conjugate Content (mg / mL) TMContent (unit / mL) Buffer Polysorbate 80 (mg / mL) Sorbitol (mg / mL) Methionine (mg / mL) Example 3-1 Trastuzumab-deruxtecan 120 200 0 10 mM succinate 0.50 45 00 1.49 Example 3-2 10 mM sodium phosphate 0.50 45 00 1.49 Example 3-3 10 mM MES 0.50 45 00 1.49 Example 3-4 10 mM sodium citrate 0.50 45 00 1.49 Example 4-1 Trastuzumab-emtansine 120 200 0 10 mM succinate 0.50 45 00 1.49 Example 4-2 10 mM sodium phosphate 0.50 45 00 1.49 Example 4-3 10 mM MES0.5045.001.49 Example 4 - 410 mM sodium citrate 0.5045.001.49 Example 5 - 1 Sacituzumab-govitecan 120200010 mM succinate 0.5045.001.49 Example 5 - 210 mM sodium phosphate 0.5045.001.49 Example 5 - 310 mM MES0.5045.001.49 Example 5 - 410 mM sodium citrate 0.5045.001.49

[0114] (1) Observation of the characteristics of pharmaceutical preparations for subcutaneous administration

[0115] The pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5 were stored under refrigerated conditions (2-8°C) for 12 weeks, and the appearance was observed by two independent workers visually checking for the presence or absence of visible particles (Table 6).

[0116] Classification Characteristics 0 Note 1 Note 2 Note 4 Note 8 Note 12 Note Example 3-1------Example 3-2------Example 3-3------Example 3-4++++++++++++++++++Example 4-1------Example 4-2------Example 4-3------Example 4-4----++Example 5-1------Example 5-2------Example 5-3------Example 5-4++++++++++++++++++

[0117] (- : No visible particles; + : Visible particles present; ++ : Many visible particles; +++ : Very many visible particles or unmeasurable due to layer separation)

[0118] Antibody-drug conjugates may exhibit stability issues such as visible particles (precipitation) and phase separation during storage. Observations of appearance revealed that, unlike when other buffers were used, the application of sodium citrate buffer resulted in visible particles being observed in all antibody-drug conjugate formulations after refrigerated storage, indicating that it is not suitable in terms of stability. In particular, a very large number of visible particles or phase separation were observed in the trastuzumab-deruxtecan formulation or the sacituzumab-govitecan formulation immediately after preparation.

[0119]

[0120] (2) pH measurement of pharmaceutical preparations for subcutaneous administration

[0121] Among the pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5, except for the case where sodium citrate buffer was applied, they were stored under refrigerated conditions (2-8℃) for 12 weeks, and the change in pH was measured with a pH meter (Table 7).

[0122] Classification Target pH pH Change 0 Note 1 Note 2 Note 4 Note 8 Note 12 Example 3-15.50 5.58 5.63 5.67 5.67 5.70 5.67 Example 3-25.67 5.72 5.76 5.74 5.77 5.76 Example 3-36.17 6.30 6.29 6.26 6.28 6.26 Example 4-15.00 5.11 5.17 5.17 5.17 5.18 5.17 Example 4-25.21 5.27 5.26 5.24 5.28 5.26 Example 4-35.56 5.64 5.65 5.62 5.58 5.50 Example 5-16.506.386.446.486.456.476.46 Example 5-26.756.786.846.826.856.83 Example 5-36.957.007.017.007.017.00

[0123] As a result of measuring pH changes, when succinate or sodium phosphate was applied as a buffer, stability was considered sufficiently ensured, as the pH change relative to the target pH was maintained within ± 0.4 for 12 weeks in all antibody-drug conjugate formulations. In particular, when succinate buffer was applied, stability was considered further enhanced, as the pH change relative to the target pH was maintained within ± 0.2 for 12 weeks in all antibody-drug conjugate formulations.

[0124] However, when MES buffer is applied, it is confirmed that the pH change relative to the target pH over 12 weeks is relatively large, so it is confirmed to be unsuitable in terms of stability.

[0125]

[0126] (3) Measurement of change in potency of natural human recombinant hyaluronidase PH20 in pharmaceutical preparations for subcutaneous administration

[0127] Among the pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5, except for the case where sodium citrate buffer was applied, they were stored under refrigerated conditions (2-8°C) for 12 weeks, and HyDIFFUZE for 12 weeks through turbidity analysis TM The change in potency was measured (Table 8).

[0128] Classification HyDIFFUZE TMChange in potency (%) 0 Note 1 Note 2 Note 4 Note 8 Note 12 Example 3-1100.0098.6495.0399.6390.6689.85 Example 3-2100.00105.04101.01104.7897.86101.35 Example 3-3100.0093.3286.1077.6957.9861.89 Example 4-1100.0085.9894.5296.3596.8790.78 Example 4-2100.0096.3096.39101.68102.4997.08 Example 4-3100.0098.6896.0098.3895.53100.36 Example 5-1100.00101.7697.9793.5995.23100.46 Example 5-2100.00102.3995.5799.5293.98101.31 Example 5-3100.00100.29100.5990.5586.8188.73

[0129] HyDIFFUZE TM As a result of measuring the change in titer, when succinate or sodium phosphate was applied as a buffer, all antibody-drug conjugate formulations did not cause a relative change in turbidity over 12 weeks, and HyDIFFUZE TM It can be seen that maintaining the potency at over 80% did not affect the activity of natural human recombinant hyaluronidase at pH 20.

[0130] However, when MES buffer was applied, the trastuzumab-deruxtecan formulation showed a significant change in turbidity starting from 4 weeks, which can be seen as a significant decrease in the activity of natural human recombinant hyaluronidase at pH 20.

[0131]

[0132] (4) Measurement of changes in antibody-drug conjugate content in pharmaceutical preparations for subcutaneous administration

[0133] Among the pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5, except for the case where sodium citrate buffer was applied, they were stored under refrigerated conditions (2-8℃) for 12 weeks, and the change in antibody-drug conjugate content over 12 weeks was measured by UV280 analysis (Table 9).

[0134] Change in content of antibody-drug conjugate (%) 0 Note 1 Note 2 Note 4 Note 8 Note 12 Example 3-1100.00105.8797.37112.7398.39105.19 Example 3-2100.0099.1997.4597.3893.9494.55 Example 3-3100.00102.6395.92100.56100.3893.33 Example 4-1100.0094.11101.53100.6798.0498.67 Example 4-2100.0098.13100.00101.9595.39101.91 Example 4-3100.0097.3799.4499.2999.74101.08 Example 5-1100.00102.2094.47100.1695.08105.20 Example 5-2100.0098.5992.9995.4693.8085.91 Example 5-3100.00101.6999.2498.9893.6095.25

[0135] As a result of measuring changes in antibody-drug conjugate content, when succinate, sodium phosphate, or MES were applied as buffers, no significant changes were observed as the content of all antibody-drug conjugates remained above 80% for 12 weeks, so all antibody-drug conjugate formulations can be considered stable preparations.

[0136]

[0137] (5) Measurement of the purity of antibody-drug conjugates in pharmaceutical preparations for subcutaneous administration

[0138] Among the pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5, except for the case where sodium citrate buffer was applied, they were stored under refrigerated conditions (2-8°C) for 12 weeks, and the change in purity of the antibody-drug conjugate over 12 weeks was measured by SEC-HPLC analysis (Table 10).

[0139] Change in purity of antibody-drug conjugate (%) 0 Note 1 Note 2 Note 4 Note 8 Note 12 Example 3-1100.0099.9299.5499.4798.1698.49 Example 3-2100.0099.9299.6499.5098.3498.71 Example 3-3100.0099.4798.5497.7796.3796.30 Example 4-1100.0099.9599.9199.5499.6299.76 Example 4-2100.00100.0099.9599.5599.5899.73 Example 4-3100.00100.0799.9599.3799.4399.66 Example 5-1100.0098.7898.2697.7596.6497.23 Example 5-2100.0099.1399.0797.8996.5297.38 Example 5-3100.0099.0998.6397.8897.2997.58

[0140] As a result of measuring changes in the purity of antibody-drug conjugates, when succinate, sodium phosphate, or MES were applied as buffers, no significant changes were observed as the purity of all antibody-drug conjugates remained within ±5% for 12 weeks, so all antibody-drug conjugate formulations can be considered stable preparations.

[0141]

[0142] (6) Measurement of changes in the drug-antibody ratio (DAR) in pharmaceutical preparations for subcutaneous administration

[0143] Among the pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5, except for the case where sodium citrate buffer was applied, they were stored under refrigerated conditions (2-8°C) for 12 weeks, and the change in DAR was measured over 12 weeks using Q-TOF and HIC-HPLC (Table 11).

[0144] Classification Theory DARDAR Change 0 Note 1 Note 2 Note 4 Note 8 Note 12 Example 3-17.707.877.867.917.897.907.89 Example 3-27.917.867.917.97.917.92 Example 3-37.897.867.897.887.897.89 Example 4-13.503.213.323.383.493.203.20 Example 4-23.353.453.453.523.353.34 Example 4-33.243.433.383.513.293.31 Example 5-17.608.007.817.927.897.787.77 Example 5-28.007.767.957.857.737.72 Example 5-38.007.787.927.867.807.76

[0145] As a result of measuring the change in DAR, it was confirmed that the change in DAR relative to the theoretical DAR was negligible for all antibody-drug conjugate formulations over 12 weeks when succinate, sodium phosphate, or MES was applied as the buffer, so stability can be considered to have been secured.

[0146]

[0147] (7) Viscosity measurement in pharmaceutical preparations for subcutaneous administration

[0148] Among the pharmaceutical formulations for subcutaneous administration prepared according to Examples 3 to 5, viscosity was measured three times repeatedly at a temperature of 25°C using a viscometer (Rheosense m-VROC), except for the case where sodium citrate buffer was applied (Table 12).

[0149] Classification Formulation Viscosity (mPa / s) Mean Standard Deviation Example 3-13.30 90.058 Example 3-23.10 90.072 Example 3-33.23 90.162 Example 4-14.34 40.118 Example 4-24.27 60.054 Example 4-34.38 20.081 Example 5-13.31 70.128 Example 5-24.03 20.601 Example 5-34.01 20.172

[0150] As a result of viscosity measurement, in all cases where succinate, sodium phosphate, or MES is applied as a buffer, the viscosity can be maintained at 1 mPa / s to 25 mPa / s (preferably 1 mPa / s to 12 mPa / s, more preferably 1 mPa / s to 6 mPa / s) under a temperature condition of 25°C, so it can be considered desirable as a pharmaceutical formulation for subcutaneous administration.

[0151] In other words, by maintaining the viscosity range as described above, a gelatin-like appearance may not be observed at all, and when used as a pharmaceutical preparation for subcutaneous administration, it may not cause side effects such as the possibility of tissue damage at the injection site, pain, or improper drug distribution.

[0152]

[0153] Synthesizing the findings shown in Tables 5 to 12, HyDIFFUZE in antibody-drug conjugates TM A pharmaceutical formulation for subcutaneous administration containing [the substance] can not only maintain a viscosity suitable for subcutaneous administration, but also maintain a constant content, purity, and DAR of the antibody-drug conjugate when stored for 12 weeks under refrigerated conditions (2-8℃), thus ensuring stability.

[0154] In particular, HyDIFFUZE in antibody-drug conjugates TMPharmaceutical formulations using succinate or sodium phosphate as a specific buffer, upon addition, can significantly improve the transparency and stability of antibody-drug conjugates provided for subcutaneous administration when stored under refrigerated conditions (2-8°C) for 12 weeks, do not cause significant changes relative to the target pH for each antibody-drug conjugate, and HyDIFFUZE TM It offers the advantage of maintaining high potency, which can be seen as further enhancing stability, suggesting that it is suitable for non-clinical and clinical applications.

[0155]

[0156] Experimental Example 1: Diffusion effect of trastuzumab-drug conjugate depending on the addition of natural human recombinant hyaluronidase PH20 in a trastuzumab-drug conjugate-based optimal subcutaneous formulation

[0157] (1) Diffusion effect of trastuzumab-deruxtecan depending on whether natural human recombinant hyaluronidase pH 20 is added to trastuzumab-deruxtecan-based subcutaneous formulations

[0158] First, as in Example 3-1, HyDIFFUZE to trastuzumab-deruxtecan 120 mg / mL TM An optimal subcutaneous formulation based on trastuzumab-deruxtecan was prepared by adding 2,000 unit / mL and combining succinate as a buffer, polysorbate 80 as a surfactant, sorbitol as a sugar or its derivative, and methionine as a viscosity reducer; the specific composition is shown in Table 5. Meanwhile, the same composition as in Example 3-1 was applied, but HyDIFFUZE TM A subcutaneous administration formulation with omitted was designated as Comparative Example 3-1 (Table 13).

[0159] Classification Trastuzumab-Deruxtecan Content / Single Dose HyDIFFUZE TMContent / Single Dose Comparison Example 3-1 Trastuzumab-deruxtecan 120 mg / mL - Trastuzumab-deruxtecan 84 mg / kg - Example 3-1 Trastuzumab-deruxtecan 120 mg / mL HyDIFFUZE TM 2,000 units / mLTrastuzumab-deruxtecan 84 mg / kgHyDIFFUZE TM 1,400 unit / kg

[0160] Subsequently, a single subcutaneous injection was administered to SD rats. Approximately 0.5 mL of blood was collected from the jugular vein using a disposable syringe (1 mL, 25 G) at 0 hours prior to administration, 2 and 7 hours after administration, and on days 1, 2, 3, 4, 7, 10, 14, 21, and 28. The collected blood was placed in an EDTA tube and centrifuged at 12,000 rpm for 3 minutes to separate the plasma. A Trastuzumab ELISA kit (Matriks biotek, Cat. No. TRA-FD-HH) was used to analyze the quantification of Trastuzumab in SD rat plasma. Antibody standard solutions were serially diluted 1 / 3 with Assay buffer to achieve a calibration range of 1.23–300 ng / mL, and samples were prepared by diluting them in Assay buffer to ensure they could be measured within the calibration range. The experiment was performed according to the ELISA kit analysis procedure, and the absorbance at 450 nm was measured using a microplate reader. Based on the analysis results, pharmacokinetic analysis was performed using the WinNonlin program (Table 14 and Fig. 3(a)).

[0161] Classification AUC last (ug·hr / mL)C max (ug / mL)T max (hr)t 1 / 2 (hr)CL t(mL / hr / kg)Vd(mL / kg)MRT(hr) Comparative Example 3-1 Average 150858.2388.696.0178.10.00050.13247.6 Standard Deviation 3456.532.963.568.30.00000.0417.9N3333333 Example 3-1 Average 183318.7541.848.0185.10.00040.11242.2 Standard Deviation 23487.571.941.629.80.00010.0110.6N3333333 Example 3-1 of Comparative Example 3-1 Percentage(%) 121.5 213 9.4 250.00 104.05 100.00 84.6 297.82

[0162] As a result, the content / single dose of trastuzumab-deruxtecan is equivalent and HyDIFFUZE TM In the case of Example 3-1, which is the optimal subcutaneous administration formulation with added HyDIFFUZE TM Compared to Comparative Example 3-1, a subcutaneous administration formulation without added, AUC last and C max It is confirmed to be at levels of approximately 121% and approximately 139%. That is, HyDIFFUZE TM In the case of the optimal subcutaneous formulation with added , trastuzumab diffusion can be seen to increase by about 21% compared to when it is not added.

[0163] In particular, as Example 3-1 is formulated with a high concentration of trastuzumab-deruxtecan at 120 mg / mL, the AUC of Example 1, which previously reviewed trastuzumab-deruxtecan at 40 mg / mL, last and C max It is confirmed that this represents a significantly increased level compared to the levels of approximately 106% and 126% compared to Comparative Example 1. Therefore, the drug transfer amount (AUC last and C max It can be seen as having maximized ).

[0164]

[0165] (2) Trastuzumab-emtansine diffusion effect depending on whether natural human recombinant hyaluronidase pH 20 is added to the optimal subcutaneous formulation based on trastuzumab-emtansine

[0166] Next, as in Example 4-1, HyDIFFUZE to trastuzumab-emtansine 120 mg / mL TM An optimal subcutaneous formulation based on trastuzumab-emtansine was prepared by adding 2,000 unit / mL and combining succinate as a buffer, polysorbate 80 as a surfactant, sorbitol as a sugar or its derivative, and methionine as a viscosity reducer; the specific composition is shown in Table 5. Meanwhile, the same composition as in Example 4-1 was applied, but HyDIFFUZE TM A subcutaneous administration formulation with omitted was designated as Comparative Example 4-1 (Table 15).

[0167] Classification: Trastuzumab-Emtansine Content / Single Dose HyDIFFUZE TM Content / Single Dose Comparison Example 4-1 Trastuzumab-emtansine 120 mg / mL - Trastuzumab-emtansine 84 mg / kg - Example 4-1 Trastuzumab-emtansine 120 mg / mL HyDIFFUZE TM 2,000 units / mLTrastuzumab-emtansine 84 mg / kgHyDIFFUZE TM 1,400 unit / kg

[0168] Subsequently, pharmacokinetic analysis was performed using the same method as in (1) (Table 16 and Fig. 3(b)).

[0169] Classification AUC last (ug·hr / mL)C max (ug / mL)T max (hr)t 1 / 2 (hr)CL t(mL / hr / kg)Vd(mL / kg)MRT(hr) Comparative Example 4-1 Average 123934.76523.14120.0049.280.00070.05167.39 Standard Deviation 22595.3079.1041.574.070.00010.018.56 N3333333 Example 4-1 Average 154922.73635.0088.0039.040.00050.03162.34 Standard Deviation 9851.6161.1213.861.420.00000.003.42 N3333333 Example 4-1's relative to Comparative Example 4-1 Percentage(%)125.00121.3873.3379.2271.4360.0096.98

[0170] As a result, the trastuzumab-emtansine content / single dose is equivalent and HyDIFFUZE TM In the case of Example 4-1, which is the optimal subcutaneous administration formulation with added HyDIFFUZE TM Compared to Comparative Example 4-1, a subcutaneous administration formulation without added, AUC last and C max It is confirmed to be at the level of approximately 125% and approximately 121%. That is, HyDIFFUZE TM In the case of the optimal subcutaneous formulation with added , trastuzumab diffusion can be seen to increase by about 25% compared to cases without it.

[0171] In other words, in the case of trastuzumab-based antibody-drug conjugates, it is proven that the diffusion effect of the trastuzumab-drug conjugate is excellent upon the addition of natural human recombinant hyaluronidase PH20, regardless of the type of conjugated drug. Furthermore, when the trastuzumab-based antibody-drug conjugate is concentrated to a high concentration of 50 mg / mL or higher, HyDIFFUZE in the optimal subcutaneous administration formulation TM It can be seen that the synergistic effect resulting from the addition is even more pronounced.

[0172]

[0173] Experimental Example 2: Diffusion effect of sacituzumab-drug conjugate depending on the addition of natural human recombinant hyaluronidase PH20 in sacituzumab-drug conjugate-based subcutaneous formulations

[0174] First, as in Example 5-1, HyDIFFUZE in sacituzumab-govitecan 120 mg / mL TM An optimal subcutaneous formulation based on sacituzumab-govitecan was prepared by adding 2,000 unit / mL and combining succinate as a buffer, polysorbate 80 as a surfactant, sorbitol as a sugar or its derivative, and methionine as a viscosity reducer; the specific composition is shown in Table 5. Meanwhile, the same composition as in Example 5-1 was applied, but HyDIFFUZE TM A subcutaneous administration formulation with omitted was designated as Comparative Example 5-1.

[0175] Next, an optimal subcutaneous formulation based on sacituzumab-govitecan was prepared by applying the same composition as in Example 5-1 but increasing the sacituzumab-govitecan content to 150 mg / mL, and this was designated as Example 6-1. Meanwhile, applying the same composition as in Example 6-1, HyDIFFUZE TM A subcutaneous administration formulation with omitted was designated as Comparative Example 6-1 (Table 17).

[0176] HyDIFFUZE sacituzumab-govitecan content / single dose TM Content / Single Dose Comparison Example 5-1 Sacituzumab-govitecan 120 mg / mL - Sacituzumab-govitecan 84 mg / kg - Example 5-1 Sacituzumab-govitecan 120 mg / mL HyDIFFUZE TM 2,000 units / mLSacituzumab-govitecan 84 mg / kgHyDIFFUZE TM1,400 unit / kg Comparative Example 6-1Sacituzumab-govitecan 150 mg / mLSacituzumab-govitecan 84 mg / kgExample 6-1Sacituzumab-govitecan 150 mg / mLHyDIFFUZE TM 2,000 units / mLSacituzumab-govitecan 84 mg / kgHyDIFFUZE TM 1,120 unit / kg

[0177] Subsequently, pharmacokinetic analysis was performed in the same manner as in (1) of Experimental Example 1, except that a Sacituzumab ELISA Kit (Abbexa, Cat. No abx395055) was used (Table 18 and Fig. 4(a)-(b)).

[0178] Classification AUC last (ug·hr / mL)C max (ug / mL)T max (hr)t 1 / 2 (hr)CL t(mL / hr / kg)Vd(mL / kg)MRT(hr) Comparative Example 5-1 Average 41354.99303.3748.00119.380.00200.34150.04 Standard Deviation 3924.5763.430.000.620.00020.032.35 N3333333 Example 5-1 Average 50144.38398.6948.00130.540.00160.30131.52 Standard Deviation 4857.3050.860.0036.560.00010.069.74 N3333333 Example 5-1 of Comparative Example 5-1 Percentage (%) 1 2 1.25 1 3 1.4 2 1 0 0.00 1 0 9.3 5 8 0.00 8 8.2 4 8 7.66 Comparative Example 6-1 Average 3 7 7 2 7.6 5 2 3 5.4 2 6 4.00 1 2 7.4 7 0.00 2 1 0.3 9 1 5 6.16 Standard Deviation 3 0 4 8.8 6 6 5.2 1 3.8 6 3 8.6 3 0.00 2 0.09 1 1.67 N 3 3 3 3 3 Example 6-1 Average 4 7 5 1 1.8 8 3 5 1.3 5 6.00 2 1 3.3 5 0.00 1 7 0.5 3 1 3 3.97 Standard Deviation 1 7 0 7.0 3 4 5.3 1 1 3.8 6 1 1 3.7 0 0.00 0 0.2 8 3.62 N 3 3 3 3 3 Example for Comparative Example 6-1 6-1 percentile (%) 125.93 149.24 87.50 167.37 80.95 135.90 85.79

[0179] As a result, HyDIFFUZE at a high concentration of sacituzumab-govitecan content of 120-150 mg / mL TM In the case of Example 5-1 or 6-1, which is the optimal subcutaneous administration formulation containing HyDIFFUZE TM Compared to Comparative Examples 5-1 or 6-1, which are subcutaneous formulations without added, the drug transfer amount (AUC last and C max As it is confirmed that all ) increase, the drug diffusion effect is judged to be excellent. In particular, HyDIFFUZE in a state where the sacituzumab-govitecan content is concentrated to a significantly high level of 150 mg / mL TM In the case of Example 6-1, which is the optimal subcutaneous administration formulation with added, the AUC compared to Comparative Example 6-1 last and C maxDrug transfer amounts (AUC) at levels of approximately 125% and 149% last and C max It is confirmed that ) is increasing further.

[0180]

[0181] Synthesizing the findings in Tables 13 to 18, HyDIFFUZE in highly concentrated antibody-drug conjugates (antibody-drug conjugate content: ≥ 50 mg / mL) TM When added, it can be considered effective as it can maximize the diffusion effect of the antibody-drug conjugate in the optimal subcutaneous administration formulation. In particular, even under high concentration conditions where the antibody-drug conjugate content is 50 mg / mL or higher, as the antibody-drug conjugate content increases, HyDIFFUZE in the optimal subcutaneous administration formulation TM It can be seen that the synergistic effect resulting from the addition is even more pronounced.

[0182]

[0183] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. Antibody-drug conjugate (ADC); and A pharmaceutical preparation for subcutaneous administration containing natural human recombinant hyaluronidase PH20.

2. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the antibody in the above antibody-drug conjugate comprises one or more antibodies selected from the group consisting of trastuzumab, sacituzumab, datopotamab, gemtuzumab, inotuzumab, moxetumomab, brentuximab, poletuzumab, enfortumab, belantamab, loncastuximab, mirvetuximab, and tositumomab.

3. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, wherein the antibody-drug conjugate above is characterized in that the drug has a molecular weight of 100 g / mol to 5,000 g / mol.

4. In Paragraph 1, The above antibody-drug conjugates are Trastuzumab-deruxtecan, Trastuzumab-emtansine, Sacituzumab-govitecan, Datopotamab-deruxtecan, Gemtuzumab-ozogamicin, Inotuzumab-ozogamicin, Moxetumomab-pasudotox, Brentuximab-vedotin, Polatuzumab-vedotin, Enfortumab-vedotin, and Belantab-mafodotin, A pharmaceutical preparation for subcutaneous administration characterized by comprising one or more selected from the group consisting of Loncastuximab-tesirine and Mirvetuximab-soravtansine.

5. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the antibody-drug conjugate comprises one or more selected from the group consisting of trastuzumab-deruxtecan, trastuzumab-emtansine, and sacituzumab-govitecan.

6. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the content of the antibody-drug conjugate is 20 mg / mL to 240 mg / mL.

7. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the content of the antibody-drug conjugate is 50 mg / mL to 240 mg / mL.

8. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the above-described natural human recombinant hyaluronidase PH20 is for the diffusion of the above-described antibody-drug conjugate.

9. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized in that the content of the above-mentioned natural human recombinant hyaluronidase PH20 is 1,000 unit / mL to 10,000 unit / mL.

10. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration characterized in that, when the above-mentioned pharmaceutical preparation for subcutaneous administration is administered subcutaneously, the area under the concentration-time curve (AUC) of the antibody in plasma is 105% or more compared to when a preparation containing the antibody-drug conjugate alone is administered subcutaneously at the same dose.

11. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration characterized by further comprising, as a buffer, succinic acid or a salt thereof; or sodium phosphate.

12. In Paragraph 1, A pharmaceutical preparation for subcutaneous administration, characterized by further comprising, as a buffer, 1 mM to 20 mM succinic acid or a salt thereof; or sodium phosphate.

13. In Paragraph 1, As a surfactant, one or more selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and poloxamer 188; As a sugar or a derivative thereof, one or more selected from the group consisting of sorbitol, mannitol, trehalose and sucrose; and A pharmaceutical preparation for subcutaneous administration characterized by additionally comprising one or more selected from the group consisting of methionine, arginine, proline, glycine, and histidine as viscosity reducing agents.

14. In Paragraph 1, As a surfactant, polysorbate 80 in an amount of 0.05 mg / mL to 5 mg / mL; As a sugar or its derivative, sorbitol in an amount of 10 mg / mL to 100 mg / mL; and A pharmaceutical preparation for subcutaneous administration, characterized by additionally containing 0.5 mg / mL to 20 mg / mL of methionine as a viscosity reducing agent.

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