WO2020071876A1 - Trastuzumab à concentration élevée ayant une viscosité réduite ou agent de stabilisation de fragment de liaison à l'antigène associé - Google Patents

Trastuzumab à concentration élevée ayant une viscosité réduite ou agent de stabilisation de fragment de liaison à l'antigène associé

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Publication number
WO2020071876A1
WO2020071876A1 PCT/KR2019/013082 KR2019013082W WO2020071876A1 WO 2020071876 A1 WO2020071876 A1 WO 2020071876A1 KR 2019013082 W KR2019013082 W KR 2019013082W WO 2020071876 A1 WO2020071876 A1 WO 2020071876A1
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formulation
trastuzumab
antigen
binding fragment
pharmaceutical formulation
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Korean (ko)
Inventor
문효정
강정훈
정영석
김동규
홍자혜
장하라
남명주
이재민
이헌주
김용국
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Samsung Bioepis Co Ltd
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Samsung Bioepis Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions

Definitions

  • the present invention relates to a stable pharmaceutical formulation comprising trastuzumab or an antigen-binding fragment thereof, and more particularly, to a formulation having a low viscosity and high stability and a method for manufacturing the same.
  • trastuzumab or an antigen-binding fragment thereof that satisfies a dosage amount without degradation of subcutaneous tissue and has a limited volume of subcutaneous injection.
  • highly concentrated trastuzumab or antigen-binding fragment formulations thereof tend to form highly viscous solutions due to the intermolecular interaction and macromolecular properties. Due to the increase in viscosity, trastuzumab or an antigen-binding fragment thereof is difficult to manufacture, transport, store and administer.
  • problems such as aggregation or particle formation of trastuzumab or an antigen-binding fragment thereof may occur. Therefore, it is essential to develop a formulation capable of stabilizing the highly concentrated antibody trastuzumab or antigen-binding fragment thereof in various stress situations.
  • One aspect of the present invention provides a pharmaceutical formulation comprising a stable trastuzumab or antigen binding fragment thereof having low viscosity and high stability.
  • Another aspect of the invention provides a method for preparing the formulation.
  • One aspect of the present invention (a) 100 mg / ml or more of trastuzumab or an antigen-binding fragment thereof; (b) buffering agents; (c) one or more selected from the group consisting of arginine, methionine and benzoate as stabilizers; And (d) provides a stable pharmaceutical formulation comprising a surfactant.
  • a step of preparing a mixed solution by adding at least one selected from the group consisting of arginine, methionine and benzoate as a buffering agent and a stabilizing agent to a solvent, wherein at least 100 mg / ml tra is added to the mixed solution.
  • a method for preparing a stable pharmaceutical preparation comprising adding stuuzumab or an antigen-binding fragment thereof, and adding a surfactant.
  • a stable pharmaceutical formulation according to an aspect of the present invention a high concentration of trastuzumab or antigen binding thereof suitable for administration as an injection without containing a protein enzyme such as hyaluronidase, which may cause side effects in an individual after injection It was found to contain fragments at low viscosity and to improve the stability of trastuzumab or antigen binding fragments thereof.
  • the formulation is a stable pharmaceutical formulation comprising a high concentration of trastuzumab antibody or antigen-binding fragment thereof, it can be used not only for injection but also as various dosage forms.
  • 3A, 3B, and 3C are graphs showing the results of analyzing the charge change of the formulations of Example 1 and Comparative Example 1 with icIEF under temperature stress conditions.
  • Figure 4 is a graph showing the analysis of IgG changes in the formulations of Example 1 and Comparative Example 1 under temperature stress conditions by CE-SDS (NR).
  • Example 5 is a graph showing the change in the HER2 binding rate of the formulations of Example 1 and Comparative Example 1 under temperature stress conditions.
  • trastuzumab (5 mM histidine buffer (pH 6.0), 60 mM trehalose, 0.04% polysorbate 20, 25 mg / mL trastuzumab, Samsung Bioepis Inc.) and buffering agent and stability excluding surfactant
  • the topic was added to sterile distilled water in a 5L biotainer container made of polycarbonate to prepare a 5L buffer solution.
  • 3 ml of 40 mg / mL trastuzumab solution in 5 mM histidine buffer at pH 6.0 was placed in a dialysis cassette (MWCO 10 kDa) (Slide-A-Lyzer cassette, Thermo Fisher Scientific), and then the cassette containing this trastuzumab solution was The dialysis was performed in a 2L beaker containing 1,600 mL of a buffer solution, and the 5 mM histidine buffer at pH 6.0 was exchanged with the buffer solution. Thereafter, trastuzumab was concentrated to 290 mg / mL.
  • MWCO 10 kDa Slide-A-Lyzer cassette, Thermo Fisher Scientific
  • Example 1 Example 2 Example 3 Comparative Example 1 Trastuzumab (mg / mL) 270 270 270 270 Buffering agent (mM) Histidine 10 10 50 5 Stabilizer (mM) Arginine 135 135 100 - Methionine - 10 - Trehalose - 60 Sodium benzoate 100 Surfactant (w / v%) Polysorbate 20 0.04 0.04 0.04 0.04 0.04 0.04 0.04 0.04 0.04 0.04
  • Examples 11 and 12 were prepared by the following method. Polycarbonate material of buffer and stabilizer except trastuzumab (5 mM histidine buffer (pH 6.0), 60 mM trehalose, 0.04% polysorbate 20, 25 mg / mL trastuzumab, Samsung Bioepis) and surfactant A 2L buffer solution was prepared by adding it to sterile distilled water in a 2L biotainer container. 400 mL of 40 mg / mL trastuzumab solution in 5 mM histidine buffer at pH 6.0 was placed in a labscale TFF system (Merck Millipore, USA), and then used Pellicon 3 Ultracel (Cat No.
  • trastuzumab 5 mM histidine buffer (pH 6.0), 60 mM trehalose, 0.04% polysorbate 20, 25 mg / mL trastuzumab, Samsung Bioepis
  • a 2L buffer solution was prepared by adding it to sterile distilled water in a 2L bio
  • Test Example 1 Comparison of viscosity according to the combination of buffering agent and stabilizer
  • the viscosity of the formulation was measured using mVROC TM from Rheosense. 200 uL of the formulation was contained in a syringe of 250 uL, and viscosity was measured using a chip (C chip) with a sensor capable of measuring a range of 10 to 14,000 cP. In order to prevent interference between samples, the viscosity was measured using a sample of 150 uL remaining after each sample was flowed through 50 uL. All measurements were performed at 25 ° C. and measured at a shear rate between 100 and 10,000.
  • Example 2 The results are shown in Table 2 and FIG. 1.
  • the mean and standard deviation (SD) were calculated by performing 3 times for each formulation, and Table 2 shows the average per item for each formulation and the standard deviation in the lower row. It was confirmed that the viscosity of Examples 1 and 2 including the combination of histidine and arginine decreased by 91.6 cP and 81.7 cP, respectively, compared to Comparative Example 1. It was confirmed that Example 3 containing sodium benzoate had a lower viscosity than Comparative Example 1 and Examples 1 and 2.
  • Example 1 271.3 (SD: 3.78) 5.5 ⁇ 0.2 72.6 (SD: 7.8) 366 (SD: 52.2)
  • Example 2 271.2 (SD: 6.31) 62.7 (SD: 9.8) 373 (SD: 31.13)
  • Example 3 272.5 (SD: 5.0) 55.5 (SD: 4.1) 370 (SD: 0.4) Comparative Example 1 275.8 (SD: 4.33) 154.3 (SD: 8.1) 216 mOsm / kg (SD: 49.1)
  • Test example 2 Benzoate Viscosity change with concentration
  • benzoate reduced the viscosity of the formulation containing high concentration trastuzumab.
  • the effect of benzoate on the viscosity change of the formulation containing high concentration of trastuzumab was confirmed by varying the concentration of benzoate.
  • Formulations comprising benzoate at different concentrations with high concentration trastuzumab contain 0.020 w / v% PS20 and have the compositions of Examples 4-7 in Table 3.
  • the target pH of the formulations of Examples 4 to 7 is 5.5.
  • Example 4-7 was prepared in the same manner as in the preparation of the formulation of Example 3, except that the concentration of sodium benzoate and / or the composition of the stabilizer were varied.
  • Example 7 The viscosity measurement results for the formulations of Examples 3 to 7 are shown in Table 3 below.
  • the average and standard deviation were obtained by performing 3 times for each formulation, and the average for each formulation in Table 3 and the standard deviation are shown in the row below. However, in the case of Example 7, it was performed once per formulation. It was confirmed that benzoate had a viscosity significantly lower than that of Comparative Example 1, that is, 154.3 cP, in the entire range of 25 mM to 200 mM tested.
  • Formulation Trastuzumab concentration (mg / mL) Buffering agent Stabilizer Sodium benzoate Viscosity (cP) Osmotic pressure (mOsm / kg)
  • Example 4 270 50 mM His 100 mM Arg 25 mM 76.4 (SD: 2.3) 438 (SD: 5.4)
  • Example 5 270 50 mM His 50 mM 68.2 (SD: 5.1) 502 (SD: 7.8)
  • Example 6 270 50 mM His 75 mM 57.7 (SD: 1.6) 554 (SD: 9.3)
  • Example 3 270 50 mM His 100 mM 55.5 (SD: 4.1) 370 (SD: 0.4)
  • Example 7 270 50 mM His - 200 mM 30.1 543.5
  • Test example 3 SE- HPLC Stability comparison under stress conditions
  • HMW high molecular weight
  • LMW low molecular weight
  • the mobile phase was measured at the flow rate, and the temperature of the sample was maintained at 5 ⁇ 3 ° C, and the injection amount was analyzed as 100 ⁇ g of protein
  • the injection run time of the sample was 36 minutes
  • the mobile phase was pH 6.8 and 100 mM sodium phosphate , And 200 mM shodium chloride-containing buffer was used and analyzed using absorbance at 280 nm
  • HMW high molecular weight
  • LMW monomer or low molecular weight
  • Test Example 4 Comparison of Stability at Temperature Stress Using icIEF It was confirmed by using icIEF (Imaging Capillary Isoelectric Focusing) that the formulation of the Example can maintain stability while containing a high concentration of antibody under temperature stress conditions.
  • Example 1 and Comparative Example 1 prepared above 1 mL of each of the preparations of Example 1 and Comparative Example 1 prepared above was placed in a 1.5 mL microtube (Axygen) made of polypropylene, and the tube was temperature of 40 ° C. in a stability thermostat (JEIO TECH). They were exposed to stress conditions for 4 weeks. Specifically, the tube was placed in the stability thermostat and placed for 4 weeks under conditions of temperature 40 ⁇ 2 ° C. and relative humidity 75 ⁇ 5%. Charge variation value, that is,% acidification value, was measured using ICE3 (Protein Simple, USA) for the formulation stored for 4 weeks.
  • ICE3 Protein Simple, USA
  • the measurement sample is 2.9 uL of the formulation of Example 1 and Comparative Example 1, which is 1% (w / v) methylcellulose solution (101876, Protein Simple), pI marker 8.18 (Protein Simple, USA), 9.50 (Sigma, USA) , And a mixture of a suitable Pharmalyte (GE Healthcare) containing the pI section of trastuzumab was injected into the iCE3 system.
  • a suitable Pharmalyte GE Healthcare
  • Example 1 has a more constant charge than Comparative Example 1 at temperature stress.
  • Test example 5 CE-SDS ( NR Stability comparison at temperature stress using)
  • CE-SDS capillary electrophoresis sodium dodecyl sulfate
  • Example 1 and Comparative Example 1 prepared above 1 mL of each of the preparations of Example 1 and Comparative Example 1 prepared above was placed in a 1.5 mL microtube (Axygen) made of polypropylene, and the tube was temperature of 40 ° C. in a stability thermostat (JEIO TECH). They were exposed to stress conditions for 4 weeks. Specifically, the tube was placed in the stability thermostat and placed for 4 weeks under conditions of temperature 40 ⁇ 2 ° C. and relative humidity 75 ⁇ 5%. IgG, 2H1L, and other proteins were measured by CE-SDS for preparations stored for 4 weeks. After 4 weeks,% 2H1L (2 heavy and 1 light chain peptides),% other peptides and% IgG were evaluated based on the weight of the antibody tested.
  • a stability thermostat JEIO TECH
  • CE-SDS conditions were non-reduced (NR) conditions.
  • the sample was pretreated by mixing the measurement sample, SDS-MW sample buffer, internal standard, and iodo acetamide solution and heating for 5 minutes in a 70 ⁇ 2 ° C heating block.
  • 90 ⁇ L of the pre-treated sample was placed in a PCR tube.
  • the PCR tube was placed in a universal vial containing a running buffer and loaded into a PA800 plus system.
  • the cartridge temperature and sample temperature of the instrument were maintained at 25 ° C and 15 ° C, respectively.
  • Relative migration time (RMT, relative migration time) of the analytical sample is obtained from the obtained electrophoresis as the peak migration time (analyte) of the analysis sample to the peak migration time (10k Da internal standard) of the internal standard. It can be divided.
  • the CPA Corrected peak area
  • % CPA can be calculated by dividing each CPA by the sum of the CPAs of all peaks.
  • Example 1 has stability equal to or higher than that of Comparative Example 1 in temperature stress.
  • ⁇ % IgG is the% IgG value at week 4 minus the% IgG value at the starting point.
  • Test Example 6 Comparison of maintenance of activity at temperature stress It was confirmed that the formulation of the above example maintains stability under temperature stress conditions and consequently maintains activity.
  • Example 1 and Comparative Example 1 prepared above were evaluated by measuring the binding ratio to HER2 to maintain activity.
  • the HER2 binding rate was measured by a binding assay based on fluorescence resonance energy transfer.
  • Example 1 has stability equal to or higher than that of Comparative Example 1 in temperature stress, and the effect of maintaining the activity of the antibody is also improved.
  • Formulation Temperature stress (40 °C): bonding Starting point 2 weeks 4 weeks % RBA % RBA % RBA Example 1 97.00 94.00 85.00 SD: 2.08 SD: 8.72 SD: 6.00 Comparative Example 1 98.00 93.00 68.33 SD: 2.89 SD: 3.61 SD: 3.06
  • Test Example 7 Confirmation of Characteristics of Formulation Containing High Concentration of trastuzumab Even when the concentration of trastuzumab contained in the formulation was increased, it was confirmed whether the formulation of the present invention was capable of maintaining the characteristics suitable for subcutaneous or intravenous administration. If a high concentration of trastuzumab is included and the osmotic pressure and viscosity are excessively increased, the formulation may not be suitable for subcutaneous or intravenous administration.
  • Examples 8 to 12 were prepared with the composition of Table 8 by varying only the concentration of trastuzumab based on Example 1.
  • Trastuzumab used trastuzumab as in Preparation Example.
  • the viscosity was measured in the same manner as in Test Example 1.
  • Table 9 shows the results. Although Examples 1 and 8 to 12 both contained high concentrations of 200 mg / mL or higher antibody, it was confirmed that they had a low viscosity in all the Examples tested.
  • Osmolarity can be measured, for example, by a freezing point method with an Advanced osmometer 2020 (Fisher scientific, USA). Specifically, a sample of 20 ⁇ L of a high-concentration formulation diluted 1/2 with a buffer solution was measured twice by using a freezing point method. In the case of a high concentration formulation, it may not be directly measured due to an increase in viscosity and a change in the freezing point.
  • Osmolarity concentrations of the formulations of the present invention may be, for example, 200 to 400 mOsm / kg, 250 to 380 mOsm / kg, or 270 to 360 mmol / kg.
  • Examples 11 and 12 were also impossible to contain high concentration antibodies of 200 mg / mL or more and had low viscosity, and the osmolality was measured to have 355 mOsm / kg and 320 mOsm / kg.
  • the formulations of the present invention contain a high concentration of trastuzumab or antigen binding fragments of 200 mg / mL or more, but have viscosity and osmotic pressure suitable for subcutaneous or intravenous administration.
  • Test Example 8 Confirmation of the Stability of the Formulation Containing High Concentration of Trastuzumab or an Antigen-binding Fragment thereof Measured using HPLC, and icIEF, and evaluated through HER2 binding level.
  • Formulation Target pH Formulation SE-HPLC icIEF % HMW (start point) % HMW (F / T 5) % HMW (0 rpm, 72 h) % HMW (400 rpm, 72 h) % Acidity (starting point) % Acidity (F / T 5) % Acidity (0 rpm, 72 h) % Acidity (400 rpm, 72 h)
  • Example 11 5.5 ⁇ 0.2 10 mM His, 135 mM Arg, 20 mM Met0.04% PS 20 1.00 [SD: 0.03] 1.06 [SD: 0.04] 1.18 [SD: 0.05] 1.22 [SD: 0.04] 39.17 [SD: 0.29] 39.86 [SD: 0.24] 39.84 [SD: 0.50] 39.78 [SD: 0.50]
  • Example 12 0.93 [SD: 0.04] 1.02 [SD: 0.02] 1.11 [SD: 0.04] 1.13 [SD: 0.05] 39.11 [SD: 0.71] 3
  • Formulation Target pH Formulation HER2 binding % RBA (start point) % RBA (5 °C / 2 Wk) % RBA (40 °C / 2 Wk)
  • Example 11 5.5 ⁇ 0.2 10 mM His, 135 mM Arg, 20 mM Met0.04% PS 20 99.3 [SD: 14.5] 94.0 [SD: 5.3] 92.3 [SD: 9.1]
  • Example 12 93.0 [SD: 10.4] 93.3 [SD: 1.2] 83.7 [SD: 1.5]
  • Test Example 9 Comparison of stability according to the concentration of the buffering agent and the stabilizer It was confirmed the stabilizing effect of the high concentration of trastuzumab or an antigen-binding fragment thereof at various combinations and concentrations of the buffering agent and the stabilizer. Stabilization was evaluated by measuring using temperature- or freeze-thaw stress conditions and then using SE-HPLC and icIEF.
  • Example 13 The combinations and concentrations of the tested buffers and stabilizers are shown in Table 13, and were measured using SE-HPLC after applying stress as in Test Examples 3 and 4. In addition, the viscosity was measured in the same manner as described in Test Example 1. As a result, the viscosity at the start of Examples 20 to 27 was 66.6, 70.2, 81.1, 73.9, 68.7, 66.3, 80.5, and 72.2 cP, respectively. Examples 20-27 included trastuzumab and PS20, their concentrations were 270 mg / mL and 0.04 w / v%, respectively. Table 13 shows the results.
  • Example 20 5.0 20 100 2.43 -3.02 +1.65 -29.13 +0.02
  • Example 21 5.0 20 200 2.79 -2.08 -2.67 -28.38 -0.03
  • Example 22 6.0 0 100 2.57 -3.12 +12.35 -30.32 -0.03
  • Example 23 6.0 20 100 2.20 -2.62 +23.96 -33.75 -0.05
  • Example 24 6.0 0 200 1.90 -2.36 +12.78 -30.20 -0.04
  • Example 25 6.0 20 200 1.81 -2.32 +24.00 -34.55 -0.10
  • Test Example 10 Comparison of stability and characteristics of formulations containing benzoate
  • the tested buffering agents, stabilizers and benzoate concentrations are as shown in Tables 14 to 17, and were measured using SE-HPLC and icIEF after applying stress as in Test Examples 3 and 4. In addition, viscosity and osmotic pressure were measured as in Test Example 7.
  • the formulations of Examples 28-31 included trastuzumab and PS20, the concentrations of which were 270 mg / mL and 0.04 w / v%, respectively. Examples 28 to 31 were adjusted to pH at pH 6.0 to 7.0. In addition, because there may be errors in manufacturing and measurement, the target pH ⁇ 0.2 was set as acceptance criteria. The median value is 6.5.
  • Table 14 shows the viscosity and osmotic pressure measured at the time point before exposure to the temperature stress condition of 40 °C for 4 weeks.
  • Table 15 shows the results measured using SE-HPLC at temperature stress.
  • Table 16 shows the results measured using icIEF at temperature stress.
  • Table 18 shows the results measured using SE-HPLC in freeze-thaw stress.
  • Example 28 10.0 30.0 75.0 20.0 6.02 277.5 336 72.1
  • Example 29 50.0 30.0 75.0 0.0 6.01 277.9 372 71.3
  • Example 30 10.0 100.0 75.0 0.0 5.95 275.5 410 76.8
  • Example 31 50.0 100.0 75.0 20.0 5.98 269.7 493 60.8
  • Examples 28 to 31 show the osmotic pressure and viscosity change according to the concentration change of pH, histidine, arginine, sodium benzoate, and methionine.
  • Examples 30-51 show changes in molecular size under temperature stress with varying concentrations of pH, histidine, arginine, sodium benzoate, and methionine.
  • Example 28 10.0 30.0 75.0 20.0 33.03 53.26 13.71 52.37 22.23 25.39 19.34
  • Example 29 50.0 30.0 75.0 0.0 33.17 52.86 13.98 52.37 21.51 26.12 19.20
  • Example 30 10.0 100.0 75.0 0.0 32.19 53.24 14.57 50.46 22.60 26.94 18.27
  • Example 31 50.0 100.0 75.0 20.0 32.87 53.52 13.61 53.87 21.25 24.88 21.00
  • Examples 30 to 51 show charge change under temperature stress according to changes in concentration of pH, histidine, arginine, sodium benzoate, and methionine.
  • Examples 30-51 show changes in molecular size under freeze-thaw stress with varying concentrations of pH, histidine, arginine, sodium benzoate, and methionine.
  • One aspect of the present invention (a) 100 mg / ml or more of trastuzumab or an antigen-binding fragment thereof; (b) buffering agents; (c) one or more selected from the group consisting of arginine, methionine and benzoate as stabilizers; And (d) provides a stable pharmaceutical formulation comprising a surfactant.
  • the pharmaceutical formulation may be liquid, or solid.
  • the pharmaceutical preparation may be in the form of powder, granule, tablet, capsule, suspension, emulsion, syrup, liquid, aerosol, ex, injection, transdermal or suppository.
  • the solid may be a dry one of the liquid.
  • the formulation of the present invention includes a high concentration of trastuzumab or an antigen-binding fragment thereof, but can maintain low viscosity and stability of trastuzumab or an antigen-binding fragment thereof.
  • antibody refers to a target molecule, ie, an immunoglobulin molecule consisting of a polypeptide chain capable of binding an antigen.
  • Such antibodies can be polyclonal antibodies, monoclonal antibodies, recombinant antibodies, single chain antibodies, hybrid antibodies, chimeric antibodies, humanized antibodies, fully human antibodies, antibodies with polyepitope specificity, or multispecific antibodies (e.g., dia Body).
  • antigen-binding fragment is a fragment capable of binding to an antigen in an antibody, including, but not limited to, Fab, F (ab ′) 2 or Fv, for example.
  • the antibody can be a monoclonal antibody.
  • the antigen can be a HER2 receptor.
  • Trastuzumab is a monoclonal antibody used to treat cancer, including breast cancer, and is marketed under the name Herceptin TM under one of several trade names. Trastuzumab is used for breast cancer that is positive for the desired HER2 receptor. In addition, trastuzumab can be used alone or in combination with other chemotherapeutic agents. Trastuzumab is administered by intravenous injection and subcutaneous injection. Trastuzumab works by binding to the HER2 receptor and slows cell replication. Trastuzumab is a recombinant IgG1 kappa humanized from mice that binds to the extracellular domain of human epidermal growth factor receptor protein (HER2), a whole antibody.
  • HER2 human epidermal growth factor receptor protein
  • the concentration of trastuzumab or an antigen-binding fragment thereof included in the formulation of the present invention may be 100 mg / ml or more.
  • the concentration may be 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290 or 300 mg / ml or more.
  • the concentration of the antibody can be freely adjusted within a range that does not substantially affect the stability, viscosity and desired pH of the liquid formulation of the present invention.
  • the concentration of the trastuzumab antibody or antigen-binding fragment thereof is, for example, 100-300 mg / mL, 110-300 mg / ml, 120-300 mg / ml, 130-300 mg in one embodiment (Group 1) / ml, 140-300 mg / ml, 150-300 mg / ml, 160-300 mg / ml, 170-300 mg / ml, 180-300 mg / ml, 190-300 mg / ml, 200-300 mg / mL, 210-300 mg / ml, 220-300 mg / mL, 110-290 mg / ml, 120-290 mg / ml, 130-290 mg / ml, 140-290 mg / ml, 150-290 mg / mL , 160 to 290 mg / ml, 170 to 290 mg / ml, 180 to 290 mg / ml, 190 to 290 mg
  • the concentration of trastuzumab or antigen-binding fragment thereof can be freely adjusted within a range that does not substantially affect the stability, desired viscosity and pH of the formulation of the present invention.
  • the concentration of trastuzumab or antigen-binding fragment thereof may contribute to its stability in the formulation, viscosity of the formulation suitable for administration, and maintenance of the pH of the formulation.
  • the formulations of the present invention may contain a buffering agent to maintain the desired pH for stabilizing trastuzumab or an antigen-binding fragment thereof.
  • buffer refers to a neutralizing material that minimizes the change in pH by acid or alkali, and in one embodiment, the buffering agent is histidine, phosphoric acid, citric acid, maleic acid, tartaric acid, succinic acid, citrate, acetate , Carbonate or a salt thereof, but is not limited thereto.
  • the buffering agent can be histidine
  • the concentration of histidine may be 1 to 150 mM, specifically, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM or more.
  • the concentration of the histidine in one embodiment (Group 2,5) 1 to 100 mM, 1 to 90 mM, 1 to 80 mM, 1 to 70 mM, 1 to 60 mM, 1 to 55 mM, 5 to 100 mM, 5 to 90 mM 5 to 80 mM, 5 to 70 mM, 5 to 60 mM, 5 to 55 mM, 7 to 100 mM, 7 to 90 mM, 7 to 80 mM, 7 to 70 mM, 7 to 60 mM, 7 To 55 mM, 10 to 100 mM, 10 to 90 mM, 10 to 80 mM, 10 to 70 mM, 10 to 60 mM, 10 to 55 mM, 10 to 50 mM, in other embodiments (Group 1) 1 to 40 mM, 1 to 30 mM, 1 to 25 mM, 5 to 40 mM, 5 to 30 mM, 5 to 25 mM, 10 to 20 mM, in another embodiment (Group 4) 40mM
  • the concentration of histidine can be freely adjusted within a range that does not affect the desired pH of the formulation of the present invention and does not affect the stability of trastuzumab or an antigen-binding fragment thereof, such as an antibody or antigen-binding fragment thereof. have.
  • the histidine may be used as a buffering agent.
  • the histidine may be histidine HCl.
  • One aspect of the present invention may include a stabilizer.
  • the stabilizer may be at least one selected from the group consisting of metal salts, amino acids and benzoates, but is not limited thereto.
  • the stabilizer may be one that does not contain sugar, sugar alcohol, sugar acid, polyol, or a combination thereof.
  • the sugar, sugar alcohol or sugar acid is glucose, fructose, galactose, sucrose, lactose, maltose, trehalose, fructooligosaccharide, galactoligosaccharide, metoligosaccharide, starch, glycogen, cellulose, chitin, pectin, glycerol, erythritol, Threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fusitol, iditol, inositol, bolitol, isomalt, maltitol, lactitol, maltotriitol, maltotetraitol, polyglycitol , Aldonic acid, ulonic acid, uronic acid, aldaric acid, stachyose, sorbose, xylose, ribose, myoinici
  • the stabilizer may be selected from one or more of arginine, methionine, or benzoate.
  • the formulation may not contain other stabilizers.
  • the formulation may not include sugar, sugar alcohol, sugar acid, or a combination thereof as a stabilizer.
  • the formulation may not include trehalose.
  • Arginine as used herein includes arginine monohydrochloride.
  • Arginine monohydrochloride may exist in L-form or D-form, or a mixture of the two. It is also known as (2S) -2-amino-5-[(aminoiminomethyl) amino] pentanoic acid monohydrochloride, arginine hydrochloride, and arginine ⁇ HCl.
  • Arginine monohydrochloride can be prepared by neutralizing arginine with hydrochloric acid. The concentration of arginine may be 1 to 300 mM.
  • the concentration of the arginine in one embodiment (Group 3, 5) 1 to 100 mM, 5 to 100 mM, 10 to 100 mM, 15 to 100 mM, 20 to 100 mM, 25 to 100 mM, 30 to 100 mM day
  • Group 1 100-300mM, 100-290mM, 100-280mM, 100-270mM, 100-260mM, 100-250mM, 100-240mM, 100-230mM, 100-220mM, 100-210mM , 100 to 200mM
  • Group 2 30 to 300mM, 50 to 300mM, 60 to 300mM, 70 to 300mM, 80 to 300mM, 90 to 300mM, 100 to 300mM, 110 to 300mM, 120 300 to 300 mM, 130 to 300 mM, 30 to 250 mM, 50 to 250 mM, 60 to 250 mM, 70 to 250 mM, 80 to 250 mM, 90 to 250 mM, 100 to 250 mM
  • the concentration of arginine can be freely adjusted within a range that does not affect the desired pH of the formulation of the present invention and does not affect the stability of trastuzumab or antigen-binding fragments thereof.
  • the arginine may be used as a stabilizer for increasing the stability of trastuzumab or an antigen-binding fragment thereof.
  • the concentration of benzoate may be 10 mM or more.
  • the concentration of the benzoate in one embodiment (Group 3) 10 to 200 mM, 15 to 200 mM, 20 to 200 mM, 10 to 190 mM, 15 to 190 mM, 20 to 190 mM, 10 to 180 mM, 15 to 180 mM , 20 to 180 mM, 10 to 170 mM, 15 to 170 mM, 20 to 170 mM, 10 to 160 mM, 15 to 160 mM, 20 to 160 mM, in another embodiment (Group 5) 30 to 200 mM, 40-200 mM, 50-200 mM, 60-200 mM, 70-200 mM, 30-190 mM, 40-190 mM, 50-190 mM, 60-190 mM, 70-190 mM, 30-180 mM, 40- 180 mM, 50-180 mM, 60-180 mM, 70-180 mM, 30
  • the benzoate may be prepared by reacting benzoic acid with a metal salt such as NaCl, KCl, NaF, KBr, NaBr, Na 2 SO 4 , NaSCN, or K 2 SO 4 , but is not limited thereto.
  • the benzoate may be sodium benzoate.
  • the benzoate can be freely selected within a range that does not affect the desired pH of the formulation of the invention and does not affect the stability of trastuzumab or antigen-binding fragments thereof.
  • the benzoate may be used as a stabilizer for increasing the stability of trastuzumab or an antigen-binding fragment thereof.
  • the concentration of the methionine may be 1 to 50 mM, in one embodiment, 1 to 40 mM, 1 to 30 mM, 1 to 25 mM, 1 to 20 mM, 5 to 40 mM, 5 to 30 mM, 5 to 20 mM, 8 to 40 mM, 8 to 30 mM, 8 to 20 mM, 10 to 40 mM, 10 to 30 mM.
  • the methionine may be used as a stabilizer for increasing the stability of trastuzumab or an antigen-binding fragment thereof.
  • the formulation may not include hyaluronidase.
  • Hyaluronidase includes all of a variety of commonly known hyaluronidases or modified hyaluronidases, such as the enzyme protein having the amino acid sequence of GenBank: AAC70915.1 in humans.
  • the hyaluronidase enzyme may be a glycoprotein, for example, rHuPH20.
  • the formulation of the present invention does not contain hyaluronidase, but has a low viscosity.
  • the viscosity of the formulation may be 100 cP or less, for example, 95 cP or 90 cP or less.
  • the viscosity of the formulation is 1 to 100 cP, 22 to 100 cP, 22 to 86 cP, 20 to 88 cP, 17 to 91 cP, 12 to 96 cP, 22 to 74 cP, 20 to 76 cP, 17 to 79 cP, 12 to 84 cP, 53 to 82 cP, 51 to 84 cP, 48 to 87 cP, 43 to 92 cP, about 30 cP, 28 to 32 cP, 25 to 35 cP, 20 to 40 cP, 34 To 73 cP, 32 to 75 cP, 29 to 78 cP, 24 to 83 cP, 47 to 86 cP, 45 to 88
  • the surfactant is a polyoxyethylene sorbitan fatty acid ester (Tween), polyethylene-polypropylene glycol, polyoxyethylene stearate, polyoxyethylene alkyl ether, polyoxyethylene-polyoxypropylene copolymer (poloxamer ), Or sodium dodecyl sulfate.
  • the polyoxyethylene alkyl ether may be, for example, polyoxyethylene monolauryl ether, alkylphenylpolyoxyethylene 30 ether (Triton-X), and specifically, a nonionic surfactant.
  • the nonionic surfactant is, for example, polysorbate, that is, polyoxyethylene sorbitan fatty acid ester or a derivative thereof, poloxamer, that is, polyoxyethylene-polyoxypropylene copolymer or a derivative thereof, and sorbitan ester. It may be one or more selected from the group consisting of.
  • the polysorbate can be polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, or a combination thereof.
  • the poloxamer may be poloxamer 188.
  • the surfactant has a concentration of 0.01 to 0.7w / v%, for example, 0.01 to 0.1w / v%, 0.01 to 0.09 w / v%, 0.01 to 0.08 w / v%, 0.01 to 0.07 w / v%, 0.02 to 0.06 w / v%, or 0.03 to 0.05 w / v%.
  • the pH of the formulation may be 4.0 to 7.5.
  • the pH of the formulation is 5.0 to 7.0, 4.9 to 7.1, 4.8 to 7.2, 4.7 to 7.3, 4.5 to 7.5, 5.3 to 7.0, 5.2 to 7.1, 5.1 to 7.2, 5.0 to 7.3, 4.8 to 7.5, About 5.5, 5.4 to 5.6, 5.3 to 5.7, 5.2 to 5.8, 5.1 to 5.9, 5.0 to 6.0, 4.8 to 6.2, 5.5 to 6.5, 6.0 to 7.0, 5.9 to 7.1, 5.8 to 7.2, 5.7 to 7.3, or 5.5 to May be 7.5.
  • the pH of the formulation can be freely adjusted within a range suitable for preventing aggregation and maintaining activity of trastuzumab or an antigen-binding fragment thereof.
  • Stabilizers, buffering agents, and surfactants as defined above may mean different things.
  • the trastuzumab or antigen-binding fragment thereof may be stabilized.
  • the liquid formulation may be a stable liquid pharmaceutical formulation.
  • stabilization means that trastuzumab or an antigen-binding fragment thereof substantially retains its physical stability, chemical stability and / or biological activity before and after administration, during further manufacturing processes, storage or storage. Physical stability, chemical stability and / or biological activity can be assessed by commonly known methods.
  • the trastuzumab or antigen-binding fragment thereof exhibits any signs of aggregation, precipitation and / or denaturation upon visual inspection for color and / or transparency or as measured by ultraviolet (UV) light scattering or size exclusion chromatography. If not visible, it retains physical stability within the pharmaceutical formulation.
  • UV ultraviolet
  • “Stability” can be measured for a selected period of time at a selected temperature.
  • a stable formulation is one in which no significant change is observed at 2-8 ° C. for 12 months or longer.
  • a stable formulation is one that does not exhibit significant changes at 2-8 ° C. for 18 months or longer.
  • a stable formulation is one where no significant change is observed for 3 months or longer at 23-27 ° C.
  • a stable formulation is one where no significant changes are observed for 6 months or longer at 23-27 ° C.
  • a stable formulation is one where no significant change is observed at 23-27 ° C. for 12 months or longer.
  • a stable formulation is one where no significant change is observed at 23-27 ° C. for 18 months or longer.
  • Stability criteria for antibody formulations are as follows. When measured by SE-HPLC, the antibody monomer is 10% or less, for example, 5% or less, or 2.5% or less relative to the initial monomer amount. For example, when measuring low molecular weight (LMW) species by SE-HPLC, the change in the amount of the low molecular weight species is less than 10%, less than 5%, or less than 2.5% relative to its initial amount. It may have.
  • LMW low molecular weight
  • the change in the amount of the high molecular weight species is 10% or less, 5% or less, or 2.5% or less relative to its initial amount. It may have.
  • the concentration of the formulation, and the pH may have a change of 10% or less, 5% or less, or 2.5% or less relative to the initial value.
  • the antibody or antigen-binding fragment thereof retains the biological activity in the pharmaceutical agent when the antibody or antigen-binding fragment thereof is biologically active for its intended purpose in the pharmaceutical preparation.
  • the biological activity is, for example, about 30%, about 20%, about 10%, about 5%, about 3 of the biological activity of the antibody or antigen-binding fragment thereof in the pharmaceutical agent at the time of manufacture of the pharmaceutical agent. % Or retain biological activity if within analytical error.
  • the biological activity can be determined, for example, in antigen binding assays.
  • the trastuzumab or antigen-binding fragment thereof retains chemical stability within the pharmaceutical agent when it is chemically stable at a predetermined time that appears to still retain biological activity. Chemical stability can be assessed by detecting and quantifying the chemically modified form of trastuzumab. Chemical modification includes size modification or charge change. The charge change can be, for example, a charge change that occurs as a result of deamidation.
  • the size modification is size exclusion chromatography (SE-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), capillary electrophoresis-sodium dodecyl sulfate, CE-SDS ) Analysis, and substrate-assisted laser desorption / ionization / time-of-flight mass spectrometry (MALDI / TOF MS).
  • charge changes can be evaluated by ion exchange chromatography (IEC), and imaged capillary isoelectric focusing (icIEF).
  • Activity can be determined through HER2 receptor binding assay.
  • the HER2 receptor binding assay can be performed by an enzyme-linked immunosorbent assay (ELISA).
  • ELISA analysis measures the ratio (%) of trastuzumab binding to the HER2 receptor.
  • ELISA can be performed using a HER2 ELISA kit (R & D system). After reacting with the trastuzumab sample and the HER2 receptor on the ELISA substrate, the degree of binding can be determined by measuring absorbance at 450 nm to obtain a relative binding rate (%).
  • Stabilization may include temperature stress, for example, 1 to 4 weeks or 8 weeks at 40 ° C; Freeze-thaw stress, eg, -70 ° C freezing and thawing at room temperature, 5 repetitions; Alternatively, it can be evaluated by measuring stirring stress, for example, by applying 400 rpm rotational force in a centrifuge for 72 hours, and then measuring% HMW,% monomer and / or% LMW using SE-HPLC.
  • the formulations of the invention may have smaller ⁇ % HMW, ⁇ % monomer, ⁇ % acidification value, and / or ⁇ % LMW value compared to Herceptin®.
  • the formulation of the present invention comprises a combination of a stabilizer, a buffering agent, and a surfactant as described above, between trastuzumab or an antigen-binding fragment thereof by interaction with the side chain of trastuzumab or an antigen-binding fragment thereof.
  • Storage stability is high because it blocks aggregation.
  • the formulation may be for injection.
  • the formulation may be for subcutaneous or intravenous injection.
  • the formulation of the present invention contains a high concentration of trastuzumab or an antigen-binding fragment thereof and is highly concentrated, but has reduced viscosity, making it easy to manufacture, transport and store trastuzumab or an antigen-binding fragment thereof, and to reduce pain during administration. Does not cause In one embodiment, the formulation of the present invention does not contain hyaluronidase, so the possibility of side effects is low.
  • the formulation may further include an aqueous carrier suitable for injection.
  • aqueous carrier is a safe and non-toxic pharmaceutical acceptable when administered to humans, including, but not limited to, sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), sterile saline solution, Ringer's solution, dextrose Does not work.
  • SWFI sterile water for injection
  • BWFI bacteriostatic water for injection
  • sterile saline solution sterile saline solution
  • Ringer's solution sterile saline solution
  • dextrose does not work.
  • the formulations of the present invention may have an appropriate range of osmolality upon subcutaneous or intravenous injection.
  • Osmolarity concentrations are, for example, 257 to 636, 285 to 517, 275 to 527, 255 to 547, 235 to 567, 257 to 485, 247 to 495, 227 to 515, 207 to 535, 290 to 608, 280 to 618, 260 to 638, 240 to 658, 200 to 400, 250 to 350, 270 to 330, 336 to 636, 326 to 646, 316 to 656, about 544, 534 to 554, or 524 to 564 mOsm / kg have.
  • the osmolarity concentration can be appropriately adjusted to minimize pain that may occur during administration.
  • the formulations of the present invention may have an appropriate range of viscosity upon subcutaneous or intravenous injection.
  • Such viscosities can include the viscosity ranges discussed above.
  • the viscosity range can be appropriately adjusted to minimize pain that may occur during administration.
  • trastuzumab or an antigen-binding fragment thereof
  • histidine as a buffering agent
  • arginine as a stabilizer
  • methionine and benzoate
  • PS 20 as a surfactant
  • the formulation has at least 150-300 mg / mL trastuzumab or an antigen-binding fragment thereof, 1-150 mM histidine as a buffering agent, 1-300 mM arginine as a stabilizer, 1-50 mM methionine and 10- It may be a stable pharmaceutical formulation comprising at least one selected from the group consisting of 220 mM benzoate, and 0.01 to 0.1 w / v% PS 20 as a surfactant.
  • the formulation comprises at least 200-290 mg / mL trastuzumab or an antigen-binding fragment thereof, 1-70 mM histidine as a buffering agent, 10-220 mM arginine as a stabilizer, 5-25 mM methionine and 15- It may be a stable pharmaceutical formulation comprising at least one selected from the group consisting of 210 mM benzoate, and 0.01 to 0.07 w / v% PS 20 as a surfactant.
  • the formulation comprises at least 220 to 270 mg / mL trastuzumab or an antigen binding fragment thereof, 10 to 50 mM histidine as a buffering agent, 30 to 200 mM arginine as a stabilizer, 10 to 20 mM methionine and 25 to It may be a stable pharmaceutical formulation comprising at least one selected from the group consisting of 200 mM benzoate, and 0.01 to 0.07 w / v% PS 20 as a surfactant.
  • the formulation may not contain sugars, sugar alcohols and sugar acids.
  • the pH of the formulation is 5.0 to 7.0, 4.9 to 7.1, 4.8 to 7.2, 4.7 to 7.3, 4.5 to 7.5, 5.3 to 7.0, 5.2 to 7.1, 5.1 to 7.2, 5.0 to 7.3, 4.8 to 7.5, about 5.5, 5.4 to 5.6, 5.3 to 5.7, 5.2 to 5.8, 5.3 to 5.7, 5.2 to 5.8, 5.1 to 5.9, 5.0 to 6.0, 4.8 to 6.2, 6.0 to 7.0, 5.9 to 7.1, 5.8 to 7.2, 5.7 to 7.3, or 5.5 to 7.5 Can be
  • the viscosity of the formulation is 100 cP or less, 90 cP, 22 to 86 cP, 20 to 88 cP, 17 to 91 cP, 12 to 96 cP, 22 to 73 cP, 20 to 75 cP, 17 to 78 cP, 12 to 83 cP, 53-82 cP, 51-84 cP, 48-87 cP, 43-92 cP, about 30 cP, 28-32 cP, 25-35 cP, 20-40 cP, 34-73 cP, 32-75 cP , 29 to 78 cP, 24 to 83 cP, 47 to 86 cP, 45 to 88 cP, 42 to 91 cP, or 37 to 96 cP.
  • the osmolality of the formulations is 257 to 636, 285 to 517, 275 to 527, 255 to 547, 235 to 567, 257 to 485, 247 to 495, 227 to 515, 207 to 535, 290 to 608, 280 to 618 , 260 to 638, 240 to 658, 200 to 400, 250 to 350, 270 to 330, 336 to 636, 326 to 646, or 316 to 656 mOsm / kg.
  • trastuzumab or an antigen-binding fragment thereof; It may be a stable pharmaceutical formulation comprising 1 to 150 mM histidine, 1 to 300 mM arginine, and 0.01 to 0.1 w / v% PS 20.
  • Trastuzumab at 190 to 300 mg / mL or an antigen binding fragment thereof; It may be a stable pharmaceutical formulation comprising 1 to 70 mM histidine and 50 to 250 mM arginine and 0.01 to 0.1 w / v% PS 20.
  • 200 to 290 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 1 to 60 mM histidine, 80 to 220 mM arginine and 0.01 to 0.1 w / v% PS 20.
  • trastuzumab at 220-270 mg / mL or an antigen-binding fragment thereof; It may be a stable pharmaceutical formulation comprising 10 to 20 mM histidine and 100 to 200 mM arginine and 0.01 to 0.1 w / v% PS 20.
  • the pH of the formulation may be between pH 5.0 and 6.0, 4.8 and 6.2, or 4.6 and 6.4.
  • the formulation may have a viscosity of 22 to 74 cP, 20 to 76 cP, or 17 to 79 cP.
  • the formulation may have an osmotic pressure of 285 to 517, 275 to 527, or 255 to 547 mOsm / kg.
  • the liquid composition is a change amount of% HMW measured when stored at 40 ° C for 4 weeks with conventional SE-HPLC, when the antibody content is 220 to 270 mg / ml (pH 5.5).
  • % HMW at Week 4-% HMW at Week 0 is about 3.2 or less, about 3.1 or less, about 3.0 or less, about 2.9 or less, about 2.8 or less, about 2.7 or less, about 2.6 or less, about 2.5 or less, about 2.4 or less , About 2.3 or less, about 2.2 or less, about 2.1 or less, about 2.0 or less, about 1.93% or less, 1.0 to 3.2%, or 1.5 to 3.1%, but is not limited thereto.
  • the "O parking" indicates initial storage.
  • the liquid formulation is the antibody content of 220 to 270 mg / ml (pH 5.5), the liquid composition is maintained at -70 ⁇ 10 °C for 18 hours, and then left at room temperature for 1 hour After repeating the thawing process 5 times, the amount of change in% HMW measured by conventional SE-HPLC is about 0.2% or less, about 0.1% or less, about 0.08% or less, about 0.06% or less, about 0.05% or less, about 0.04 %, About 0.03% or less, about 0.02% or less, about 0.01% or less, about 0.001% or less, about 0.08% or less, 0.06 to 0.10%, 0.04 to 0.12%, or 0.02 to 0.14%.
  • the liquid composition when the antibody content is 220 to 270 mg / ml (pH 5.5), is stored at 40 ° C. for 4 weeks, and then the reference standard, the analysis control and the sample are 0.0488 to 100 ⁇ g / Diluted to a concentration of mL and placed in each well of a 96-well plate, the concentration of trastuzumab was measured using Nanodrop (Thermo, USA), and then trastuzumab labeled with europium (PerkinElmer, USA). After inserting the Cy5 labeled HER2 solution, the light was shielded against a 96-well plate and incubated for 60 minutes while stirring at 25 ⁇ 1 ° C.
  • the% relative binding activity (RBA) value may be 70% or more, 75% or more, 80% or more, 85% or more.
  • Another embodiment is 150-300 mg / mL trastuzumab or an antigen-binding fragment thereof; 1 to 150 mM histidine, 1 to 300 mM arginine, 1 to 50 mM methionine; And 0.01 to 0.1 w / v% PS 20.
  • 210 to 300 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 1 to 70 mM histidine, 10 to 200 mM arginine, 1 to 30 mM methionine, and 0.01 to 0.1 w / v% PS 20.
  • trastuzumab at 220-290 mg / mL or an antigen-binding fragment thereof; It may be a stable pharmaceutical formulation comprising 5 to 30 mM histidine, 100 to 200 mM arginine, 5 to 25 mM methionine, and 0.01 to 0.1 w / v% PS 20.
  • 240 to 270 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 5 to 15 mM histidine and 100 to 150 mM arginine, 10 to 20 mM methionine, and 0.01 to 0.1 w / v% PS 20.
  • the formulation may be pH 5.3 to 5.7, 5.1 to 5.9, or 4.9 to 6.1.
  • the formulation may have a viscosity of 34 to 73, 32 to 75, or 31 to 77 cP.
  • the formulation may have an osmotic pressure of 257 to 485, 247 to 495, or 227 to 515 mOsm / kg.
  • the liquid composition is a change amount of% HMW measured when stored at 40 ° C for 4 weeks with conventional SE-HPLC, when the antibody content is 240 to 270 mg / ml (pH 5.5).
  • the "O parking" indicates initial storage.
  • the liquid formulation when the antibody content is 240 to 270 mg / ml (pH 5.5), the liquid composition is maintained at -70 ⁇ 10 ° C. for 18 hours, and then left at room temperature for 1 hour.
  • the amount of change in% HMW measured by conventional SE-HPLC is about 2.5% or less, about 2% or less, about 1.5% or less, about 1.0% or less, about 0.5% or less, about 0.1 % Or less, about 0.08% or less, about 0.01% or less, about 0.001% or less, about 0.06% or less, 0.04 to 0.08%, or 0.02 to 0.10%.
  • trastuzumab or an antigen-binding fragment thereof; It may be a stable pharmaceutical formulation comprising 1 to 150 mM histidine, 1 to 300 mM arginine, 10 to 220 mM benzoate, and 0.01 to 0.1 w / v% PS 20 as surfactant.
  • trastuzumab at 220-300 mg / mL or an antigen-binding fragment thereof; It can be a stable pharmaceutical formulation comprising 1 to 70 mM histidine and 10 to 150 mM arginine, 10 to 200 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • 250 to 290 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 5 to 55 mM histidine and 20 to 110 mM arginine, 10 to 165 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • 260 to 280 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 10 to 50 mM histidine, 30 to 100 mM arginine, 25 to 150 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • the formulation may be pH 5.3 to 7.0, 5.1 to 7.2, or 4.9 to 7.4.
  • the formulation may have a viscosity of 53 to 82, 51 to 84, 48 to 87, 43 to 92, 33 to 102 cP.
  • the formulation may have an osmotic pressure of 290 to 608, 280 to 618, 260 to 638, or 240 to 658 mOsm / kg.
  • the formulation may not contain methionine.
  • the liquid composition is a% HMW measured when stored at 40 ° C for 4 weeks with conventional SE-HPLC when the antibody content is 260 to 280 mg / mL mg / ml (pH 5.5).
  • the amount of change i.e., (% HMW at Week 4 of storage)-(% HMW at Week 0) is about 3.2% or less, about 3.1% or less, about 3.0% or less, about 2.9% or less, about 2.8% or less, about 2.7% Or less, about 2.6% or less, about 2.5% or less, about 2.4% or less, about 2.3% or less, and specifically, may be 2.2 to 3.2%, but is not limited thereto.
  • the "O parking" indicates initial storage.
  • the liquid formulation when the antibody content is 260 to 280 mg / ml (pH 5.5), the liquid composition is maintained at -70 ⁇ 10 °C for 18 hours, and then left at room temperature for 1 hour After repeating the thawing process 5 times, the amount of change in% HMW measured by conventional SE-HPLC is about 0.2% or less, about 0.1% or less, about 0.08% or less, about 0.06% or less, about 0.05% or less, about 0.04 %, About 0.03% or less, about 0.02% or less, about 0.01% or less, about 0.001% or less, about 0.08% or less, 0.06 to 0.10%, 0.04 to 0.12%, or 0.02 to 0.14%.
  • trastuzumab or antigen binding fragments thereof are 150 to 300 mg / mL trastuzumab or antigen binding fragments thereof, 1 to 150 mM histidine as a buffering agent, 10 to 220 mM benzoate as a stabilizer and 0.01 to 0.1 w / v% PS as a surfactant It can be a stable pharmaceutical formulation comprising 20.
  • trastuzumab at 220-300 mg / mL or an antigen-binding fragment thereof; It may be a stable pharmaceutical formulation comprising 1 to 70 mM histidine, 100 to 250 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • 250 to 290 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 30 to 70 mM histidine, 175 to 220 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • 260 to 280 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 40 to 60 mM histidine, 185 to 215 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • the pH of the formulation may be about 5.5, 5.4 to 5.6, 5.3 to 5.7, or 5.2 to 5.8.
  • the viscosity of the formulation may be about 30 cP, 28 to 32 cP, 25 to 35 cP, or 20 to 40 cP.
  • the osmolarity concentration of the formulation may be about 543.5, 523 to 563, 503 to 583, or 483 to 603 mOsm / kg.
  • the benzoate may be sodium benzoate.
  • the concentration of PS 20 may be 0.02 to 0.06 w / v%.
  • Another embodiment is 150-300 mg / mL trastuzumab or an antigen-binding fragment thereof; 1 to 150 mM histidine, 1 to 300 mM arginine, 1 to 30 mM methionine; It may be a stable pharmaceutical formulation comprising 10 to 220 mM benzoate and 0.01 to 0.1 w / v% PS 20.
  • 240 to 300 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 1 to 70 mM histidine and 10 to 200 mM arginine, 1 to 50 mM methionine, 10 to 200 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • 250 to 290 mg / mL trastuzumab or an antigen-binding fragment thereof may be a stable pharmaceutical formulation comprising 5 to 55 mM histidine and 20 to 150 mM arginine, 5 to 25 mM methionine, 50 to 165 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • 250 to 290 mg / mL trastuzumab or an antigen-binding fragment thereof can be a stable pharmaceutical formulation comprising 10 to 50 mM histidine and 30 to 100 mM arginine, 10 to 20 mM methionine, 75 to 150 mM benzoate, and 0.01 to 0.1 w / v% PS 20.
  • the pH of the formulation may be 5.9 to 7.0, 5.8 to 7.1, 5.7 to 7.2, 5.6 to 7.3, or 5.5 to 7.4.
  • the viscosity of the formulation may be 47 to 86 cP, 45 to 88 cP, 42 to 91 cP, or 37 to 96 cP.
  • the osmolarity concentration of the formulation may be 336 to 636, 326 to 646, or 316 to 656 mOsm / kg.
  • the benzoate may be sodium benzoate.
  • the concentration of PS 20 may be 0.02 to 0.06 w / v%.
  • the liquid composition is a% HMW measured when stored at 40 ° C for 4 weeks with conventional SE-HPLC when the antibody content is 260 to 280 mg / mL mg / ml (pH 5.5).
  • the amount of change i.e., (% HMW at week 4 of storage)-(% HMW at week 0) of about 3.3% or less, about 3.2% or less about 3.1% or less, about 3.0% or less, about 2.9% or less, about 2.8% or less , About 2.7% or less, about 2.6% or less, and specifically 3.2 to 2.6%, but is not limited thereto.
  • the "O parking" indicates initial storage.
  • the liquid formulation when the antibody content is 260 to 280 mg / ml (pH 5.5), the liquid composition is maintained at -70 ⁇ 10 °C for 18 hours, and then left at room temperature for 1 hour After repeating the thawing process 5 times, the amount of change in% HMW measured by conventional SE-HPLC is about 0.2% or less, about 0.1% or less, about 0.08% or less, about 0.06% or less, about 0.05% or less, about 0.04 %, About 0.03% or less, about 0.02% or less, about 0.01% or less, about 0.001% or less, about 0.08% or less, 0.06 to 0.10%, 0.04 to 0.12%, or 0.02 to 0.14%.
  • a buffering agent in a solvent And preparing a mixed solution by adding at least one selected from the group consisting of arginine, methionine and benzoate as a stabilizer; Adding trastuzumab or an antigen-binding fragment thereof to the mixed solution at least 100 mg / ml; And it provides a method for producing a stable pharmaceutical formulation comprising the step of adding a surfactant.

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Abstract

La présente invention concerne une formulation pharmaceutique stable comprenant du trastuzumab ou un fragment de liaison à l'antigène associé.
PCT/KR2019/013082 2018-10-04 2019-10-04 Trastuzumab à concentration élevée ayant une viscosité réduite ou agent de stabilisation de fragment de liaison à l'antigène associé Ceased WO2020071876A1 (fr)

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KR10-2018-0118506 2018-10-04

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WO2020071876A1 true WO2020071876A1 (fr) 2020-04-09

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PCT/KR2019/013082 Ceased WO2020071876A1 (fr) 2018-10-04 2019-10-04 Trastuzumab à concentration élevée ayant une viscosité réduite ou agent de stabilisation de fragment de liaison à l'antigène associé

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025158306A1 (fr) * 2024-01-25 2025-07-31 Intas Pharmaceuticals Ltd. Nouvelle formulation de phesgo à haute concentration

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150009593A (ko) * 2012-05-18 2015-01-26 제넨테크, 인크. 고농도 모노클로날 항체 제제
KR20170084369A (ko) * 2011-10-28 2017-07-19 인테그리티 바이오, 아이엔씨. 아미노산을 함유하는 단백질 제제
US20170232103A1 (en) * 2014-06-20 2017-08-17 Reform Biologics, Llc Excipient compounds for biopolymer formulations
WO2018067987A1 (fr) * 2016-10-06 2018-04-12 Amgen Inc. Formulations pharmaceutiques de protéines à viscosité réduite
WO2018116198A1 (fr) * 2016-12-23 2018-06-28 Serum Institute Of India Private Limited Procédés améliorés pour augmenter la productivité d'anticorps dans une culture de cellules de mammifère et minimiser l'agrégation pendant des processus de formulation en aval, procédés de formulation et formulations d'anticorps stables obtenus à partir de ceux-ci

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20170084369A (ko) * 2011-10-28 2017-07-19 인테그리티 바이오, 아이엔씨. 아미노산을 함유하는 단백질 제제
KR20150009593A (ko) * 2012-05-18 2015-01-26 제넨테크, 인크. 고농도 모노클로날 항체 제제
US20170232103A1 (en) * 2014-06-20 2017-08-17 Reform Biologics, Llc Excipient compounds for biopolymer formulations
WO2018067987A1 (fr) * 2016-10-06 2018-04-12 Amgen Inc. Formulations pharmaceutiques de protéines à viscosité réduite
WO2018116198A1 (fr) * 2016-12-23 2018-06-28 Serum Institute Of India Private Limited Procédés améliorés pour augmenter la productivité d'anticorps dans une culture de cellules de mammifère et minimiser l'agrégation pendant des processus de formulation en aval, procédés de formulation et formulations d'anticorps stables obtenus à partir de ceux-ci

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025158306A1 (fr) * 2024-01-25 2025-07-31 Intas Pharmaceuticals Ltd. Nouvelle formulation de phesgo à haute concentration

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