[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

WO2020063668A1 - Formulation contenant un anticorps anti-ox40, son procédé de préparation et son utilisation - Google Patents

Formulation contenant un anticorps anti-ox40, son procédé de préparation et son utilisation Download PDF

Info

Publication number
WO2020063668A1
WO2020063668A1 PCT/CN2019/107829 CN2019107829W WO2020063668A1 WO 2020063668 A1 WO2020063668 A1 WO 2020063668A1 CN 2019107829 W CN2019107829 W CN 2019107829W WO 2020063668 A1 WO2020063668 A1 WO 2020063668A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
amino acid
seq
antibody preparation
preparation according
Prior art date
Application number
PCT/CN2019/107829
Other languages
English (en)
Chinese (zh)
Inventor
汪音爵
曹魏
周凯松
Original Assignee
信达生物制药(苏州)有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 信达生物制药(苏州)有限公司 filed Critical 信达生物制药(苏州)有限公司
Priority to CN201980006061.XA priority Critical patent/CN111683681B/zh
Publication of WO2020063668A1 publication Critical patent/WO2020063668A1/fr

Links

Images

Classifications

    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants

Definitions

  • the invention relates to the field of antibody preparations. More specifically, the present invention relates to a pharmaceutical formulation comprising an antibody that specifically binds OX40 (hereinafter also referred to as "anti-OX40 antibody”) and / or an antigen-binding fragment thereof, a method for preparing the pharmaceutical formulation, and The therapeutic and / or prophylactic uses of the pharmaceutical formulation are described.
  • anti-OX40 antibody an antibody that specifically binds OX40
  • antigen-binding fragment thereof hereinafter also referred to as "anti-OX40 antibody”
  • the therapeutic and / or prophylactic uses of the pharmaceutical formulation are described.
  • Co-stimulatory molecules are a type of cell surface molecules other than antigen receptors or antigen ligands that are required for lymphocytes to respond effectively to antigens. By specifically binding to co-stimulatory ligands, lymphocyte-mediated co-stimulation reaction. Co-stimulatory molecules have been shown to enhance T cell expansion, effector function, and survival in vitro; and to enhance human T cell retention and antitumor activity in vivo.
  • OX40 (also known as CD134, TNFRSF4, and ACT35) is a costimulatory molecule and has been shown to promote costimulatory signals to T cells, leading to enhanced cell proliferation, survival, effector function, and migration (Gramaglia, et al. People, Ox-40: Ligand: a potent costimulatory molecule for the primary CD4T cell response. J Immunol. 1998; 161: 6510–6517; Gramaglia I et al. The OX40 Costimulatory Receptor Determines the development of the CDC J Immunol. 2000; 165: 3043–3050).
  • Anti-OX40 antibodies that specifically bind OX40 as OX40 agonists are described in, for example, WO 2012/027328, US Patent No. 7,959,925, PCT Publication No. WO 2006/121810, and Chinese Patent Application Nos. 201710185399.9, 201710185400.8.
  • anti-OX40 antibody preparations that can be used to treat, prevent, or delay various cancers, immune-related diseases, and T-cell dysfunction diseases.
  • Antibody formulations must be formulated not only in a manner that makes the antibody suitable for administration to a subject, but also in a manner that maintains its stability during storage and subsequent use. For example, if an antibody is not properly formulated in a liquid, the antibody in a liquid solution tends to decompose, aggregate, or undergo undesired chemical modification. The stability of the antibody in the antibody preparation depends on the buffer, stabilizer, surfactant, etc. used in the preparation.
  • Antibodies against OX40 are one example of a formulation that needs to be properly formulated to treat or prevent a disease. Although some anti-OX40 antibodies are known, there remains a need in the art for novel pharmaceutical formulations containing anti-OX40 antibodies that are sufficiently stable and suitable for administration to a subject.
  • the present invention satisfies the aforementioned needs by providing a pharmaceutical formulation containing an antibody that specifically binds to OX40.
  • the invention provides a liquid antibody formulation comprising (i) an anti-OX40 antibody or an antigen-binding fragment thereof; (ii) a buffer, (iii) a stabilizer, and (iv) a surfactant.
  • the concentration of the anti-OX40 antibody or antigen-binding fragment thereof in the liquid antibody preparation of the present invention is about 1-150 mg / mL. In another embodiment, the concentration of the anti-OX40 antibody or antigen-binding fragment thereof in the liquid antibody preparation of the present invention is about 10 mg / mL to about 100 mg / mL. In other embodiments, the concentration of the anti-OX40 antibody or antigen-binding fragment thereof in the liquid antibody preparation of the present invention is about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 mg / mL.
  • the anti-OX40 antibody is any antibody that binds to an OX40 molecule (eg, a human OX40 molecule), such as a polyclonal antibody, a monoclonal antibody, or a combination of the two.
  • OX40 molecule eg, a human OX40 molecule
  • the anti-OX40 antibody is a monoclonal antibody.
  • the anti-OX40 antibody or antigen-binding fragment thereof is an anti-OX40 antibody or an antigen-binding fragment thereof as defined herein.
  • the concentration of the buffering agent in the liquid antibody formulation of the invention is about 0.1-50 mg / ml. In one embodiment, the concentration of the buffering agent in the liquid antibody formulation of the present invention is about 1-20 mg / ml, for example, about 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5 , 7, 7.5, 8 mg / ml.
  • the buffer is selected from the group consisting of phosphate, citrate, citrate solvate, succinic acid, trimethylolaminomethane, and combinations thereof, more preferably citrate, citric acid Salt hydrates, for example, sodium citrate, sodium citrate dihydrate.
  • the concentration of the stabilizer in the liquid antibody formulation of the invention is about 10-200 mg / ml. In one embodiment, the concentration of the stabilizer in the liquid antibody formulation of the invention is about 20-100 mg / ml, such as about 30, 40, 50, 60, 70, 80, 90 mg / ml.
  • the stabilizer is selected from the group consisting of sucrose, trehalose, mannitol and combinations thereof, and more preferably sucrose.
  • the concentration of the surfactant in the liquid antibody formulation of the invention is about 0.01-5 mg / ml. In one embodiment, the concentration of the surfactant in the liquid antibody formulation of the present invention is about 0.1-2 mg / ml, such as about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 mg / ml.
  • the surfactant is a non-ionic surfactant.
  • the surfactant is, for example, pluronics, polysorbate-80, polysorbate-60, polysorbate-40, or polysorbate-20, and the like.
  • the pH of the liquid formulation is about 5.0-8.0. In one embodiment, the pH of the liquid formulation is about 5.0, 6.0, 7.0, 8.0.
  • the liquid formulation is a pharmaceutical formulation, preferably an injection, and more preferably a subcutaneous injection.
  • liquid antibody formulation of the invention comprises
  • citrate, citrate hydrate such as sodium citrate, sodium citrate dihydrate, as a buffer
  • pH of the liquid preparation is about 5.0-8.0.
  • liquid antibody formulation of the invention comprises
  • pH of the liquid preparation is about 5.0-8.0.
  • liquid antibody formulation of the invention comprises
  • pH of the liquid preparation is about 6.0-7.0.
  • the present invention provides a solid antibody preparation obtained by subjecting the liquid antibody preparation of the present invention to a curing treatment.
  • the curing treatment is performed by, for example, a crystallization method, a spray drying method, or a freeze drying method.
  • the solid antibody preparation is, for example, in the form of a lyophilized powder for injection.
  • the solid antibody preparation can be reconstituted in an appropriate solvent before use to form the reconstituted preparation of the present invention.
  • the reconstituted preparation is also a liquid antibody preparation of the present invention.
  • the appropriate vehicle is selected from water for injection, organic solvents for injection, including but not limited to oil for injection, ethanol, propylene glycol, and the like, or a combination thereof.
  • a liquid or solid formulation comprising an anti-OX40 antibody of the present invention is at about -80 ° C to about 45 ° C, such as -80 ° C, about -30 ° C, about -20 ° C, about 0 ° C, about 5 °C, about 25 °C, about 35 °C, about 38 °C, about 40 °C, about 42 °C or about 45 °C for at least 10 days, at least 20 days, at least 1 month, at least 2 months, at least 3 Months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 After at least 24 months, at least 36 months, or longer, the size of the anti-OX40 antibody or its antigen-binding fragment is reduced by no more than 10%, for example, not more than 5%, by size exclusion high performance liquid chromatography. , 4%, 3%, 2%
  • a liquid or solid formulation comprising an anti-OX40 antibody of the present invention is at about -80 ° C to about 45 ° C, such as -80 ° C, about -30 ° C, about -20 ° C, about 0 ° C, about 5 °C, about 25 °C, about 35 °C, about 38 °C, about 40 °C, about 42 °C or about 45 °C for at least 10 days, at least 20 days, at least 1 month, at least 2 months, at least 3 Months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 At least 24 months, at least 36 months, or longer, detected by non-reduced sodium lauryl sulfate capillary electrophoresis (CE-SDS) and / or reduced CE-SDS, anti-OX40 antibody Or the purity of the antigen-binding fragment
  • a liquid or solid formulation comprising an anti-OX40 antibody of the present invention is at about -80 ° C to about 45 ° C, such as -80 ° C, about -30 ° C, about -20 ° C, about 0 ° C, about 5 °C, about 25 °C, about 35 °C, about 38 °C, about 40 °C, about 42 °C or about 45 °C for at least 10 days, at least 20 days, at least 1 month, at least 2 months, at least 3 Months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 At least 24 months, at least 36 months, or longer, the cation exchange high-performance liquid chromatography (CEX-HPLC) test showed that the charge variants of the anti-OX40 antibody or its antigen-binding fragment did not change more than 10%, such as no more than 5%,
  • the anti-OX40 antibody or antigen-binding fragment thereof in the liquid formulation of the present invention is capable of high affinity, for example, at 10 -7 M or less, preferably at 10 -8 M to 10 -12 M.
  • K D specifically binds OX40, and thus mediates a co-stimulatory response to an anti-OX40 antibody or antigen-binding fragment thereof.
  • the anti-OX40 antibody or antigen-binding fragment thereof in the liquid formulation of the present invention comprises a heavy chain variable region VH and / or a light chain variable region VL, wherein
  • VH shown as SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103 and 104 CDRs of the three complementary determining regions,
  • HCDR1 selected from the amino acid sequences of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 16, and 17 SEQ ID NO: 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36 of the amino acid sequence of HCDR2, and selected from SEQ ID NO: a combination of HCDR3 in the amino acid sequence of 37, 38, 39, 40, 41, 42, 43, and 44; or
  • each of the three CDRs contains at least one and no more than 5, 4, 3, 2 or 1 amino acid changes (preferably amino acid substitutions, preferably conservative substitutions) )the sequence of;
  • LCDR1 selected from the amino acid sequence of SEQ ID NO: 45, 46, 47, 48, 49, and 50
  • LCDR2 selected from the amino acid sequence of SEQ ID NO: 51, 52, 53, 54, 55, and 56
  • a combination of LCDR3 selected from the amino acid sequences of SEQ ID NO: 57, 58, 59, 60, 61, 62, 63, 64, 65 and 66; or
  • each of the three CDRs contains at least one and no more than 5, 4, 3, 2 or 1 amino acid changes (preferably amino acid substitutions, preferably conservative substitutions) )the sequence of.
  • the anti-OX40 antibody in the liquid formulation of the present invention comprises a heavy chain variable region VH and / or a light chain variable region VL, wherein,
  • the heavy chain variable region VH comprises and is selected from SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102 , 103 and 104 have an amino acid sequence of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity or 100% identity Its composition
  • the light chain variable region VL comprises at least 90%, 91%, 92%, and an amino acid sequence selected from the group consisting of SEQ ID NO: 115, 116, 117, 118, 119, 120, 121, 122, 123, and 124, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical or 100% identical amino acid sequences or consist of them.
  • the anti-OX40 antibody in the liquid formulation of the present invention comprises a heavy chain and / or a light chain, wherein the heavy chain comprises and is selected from the group consisting of SEQ ID NO: 125, 126, 127, 128, 129 , 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154 , 155, 156, 157, 158, 159, 160, 161, and 162 have an amino acid sequence of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% Identity or consist thereof, said light chain comprising at least 90%, 91%, 92% of the amino acid sequence selected from the group consisting of SEQ ID NO: 163, 164, 165, 166, 167, 168,
  • the invention provides a delivery device comprising a liquid or solid antibody preparation of the invention as described in various embodiments of the present specification.
  • the delivery device of the present invention is provided in the form of a pre-filled syringe comprising a liquid antibody formulation or a solid antibody formulation of the present invention as described in various embodiments of this specification, such as for intravenous or intramuscular injection .
  • the invention relates to a method of delivering an anti-OX40 antibody to a subject, such as a mammal, comprising administering to the subject, such as a mammal, the subject of the invention as described in various embodiments of the present specification.
  • a step of a liquid or solid antibody preparation, said delivery being performed, for example, by a delivery device using a pre-filled syringe.
  • the invention further provides the use of a liquid antibody preparation or a solid antibody preparation of the invention for the preparation of a delivery device that activates T cells in a subject or induces T cell-mediated antitumor activity or enhances the body's immune response (such as , Pre-filled syringes) or drugs, particularly for treating a disease in a subject, such as cancer, such as lung cancer (eg, non-small cell lung cancer), liver cancer, stomach cancer, or colon cancer.
  • a delivery device that activates T cells in a subject or induces T cell-mediated antitumor activity or enhances the body's immune response (such as , Pre-filled syringes) or drugs, particularly for treating a disease in a subject, such as cancer, such as lung cancer (eg, non-small cell lung cancer), liver cancer, stomach cancer, or colon cancer.
  • the present invention further provides a subject under test by administering a liquid antibody preparation or solid antibody preparation of the invention or a delivery device (e.g., a pre-filled syringe) or a drug comprising the liquid antibody preparation or solid antibody preparation of the invention to a subject.
  • a delivery device e.g., a pre-filled syringe
  • a drug comprising the liquid antibody preparation or solid antibody preparation of the invention to a subject.
  • the present invention further provides a subject to be treated by administering the liquid antibody preparation or solid antibody preparation of the invention or a delivery device (e.g., a pre-filled syringe) or a medicament comprising the liquid antibody preparation or solid antibody preparation to the subject.
  • a delivery device e.g., a pre-filled syringe
  • a medicament comprising the liquid antibody preparation or solid antibody preparation to the subject.
  • Methods of cancer such as lung cancer (such as non-small cell lung cancer), liver cancer, stomach cancer, or colon cancer.
  • Figure 1 shows a map of the charge variant of an anti-OX40 antibody placed under heat stress for 10 days at pH 5.0.
  • Fig. 2 shows a charge variant profile of an anti-OX40 antibody placed under heat stress for 10 days at pH 6.0.
  • Figure 3 shows a charge variant profile of an anti-OX40 antibody placed under heat stress for 10 days at pH 7.0.
  • Figure 4. shows a charge variant profile of an anti-OX40 antibody placed under heat stress for 10 days at pH 8.0.
  • Fig. 5 is a graph showing a change in turbidity of an anti-OX40 antibody preparation of the present invention after being left at a temperature of 40 ° C ⁇ 2 ° C.
  • Fig. 6 is a graph showing the change in purity measured by the SEC-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C ⁇ 2 ° C.
  • FIG. 7 is a graph showing the change in the acidic component of the charge variant of the anti-OX40 antibody measured by the CEX-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C. ⁇ 2 ° C.
  • FIG. 7 is a graph showing the change in the acidic component of the charge variant of the anti-OX40 antibody measured by the CEX-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C. ⁇ 2 ° C.
  • FIG. 8 is a graph showing the main component change of an anti-OX40 antibody charge variant measured by a CEX-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C. ⁇ 2 ° C.
  • FIG. 8 is a graph showing the main component change of an anti-OX40 antibody charge variant measured by a CEX-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C. ⁇ 2 ° C.
  • FIG. 9 is a graph showing changes in the basic composition of the charge variant of the anti-OX40 antibody measured by the CEX-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C. ⁇ 2 ° C.
  • FIG. 9 is a graph showing changes in the basic composition of the charge variant of the anti-OX40 antibody measured by the CEX-HPLC method after the anti-OX40 antibody preparation of the present invention was left to stand at a temperature of 40 ° C. ⁇ 2 ° C.
  • antibody formulation refers to a preparation that is in a form that allows the biological activity of the antibody as an active ingredient to be effective, and does not contain unacceptably toxic to the subject to which the preparation will be administered Extra components. Such antibody preparations are usually sterile. "Pharmaceutically acceptable" excipients are those excipients that can be reasonably administered to a test mammal to provide an effective dose of the active ingredient used.
  • anti-OX40 antibody formulation means a combination of at least one anti-OX40 antibody as an active ingredient and at least one inactive ingredient. After the combination, the anti-OX40 antibody as an active ingredient is suitable for therapeutic or prophylactic administration to human or non-human animals.
  • the anti-OX40 antibody as an active ingredient in the preparation specifically binds to the OX40 molecule, and the resulting signalling promotes a co-stimulatory signal to T cells, leading to enhanced cell proliferation, survival, effector function, and migration.
  • the antibody preparation of the present invention may be sterile, homogeneous and / or isotonic, and the antibody preparation may be directly prepared as an aqueous liquid preparation, for example, a ready-to-use pre-filled syringe, or prepared as a lyophilized
  • the formulation is reconstituted (ie, reconstituted) by dissolution and / or suspension in a physiologically acceptable solution immediately before use.
  • the anti-OX40 antibody formulation is in the form of a liquid formulation, a lyophilized formulation, or a reconstituted formulation.
  • antibodies as active ingredients of drugs has been widely used, including products such as HERCEPTIN TM (trastuzumab), RITUXAN TM (rituximab), SYNAGIS TM (palizumab), and the like. Techniques for purifying therapeutic antibodies to pharmaceutical grade are well known in the art.
  • a "sterile” formulation is a formulation that is sterile or free or essentially free of living microorganisms and their spores.
  • lyophilized preparation refers to a composition obtained or capable of being obtained by a freeze-drying treatment of a liquid preparation. Preferably, it is a solid composition having a water content of less than 5%, preferably less than 3%.
  • reconstituted preparation refers to a liquid preparation obtained by dissolving and / or suspending a solid preparation (eg, a lyophilized preparation) in a physiologically acceptable solution.
  • the anti-OX40 antibody formulations of the present invention exhibit low to undetectable levels of antibody aggregation or degradation or chemical modification during manufacturing, preparation, transportation, and long-term storage, so that there is little or no antibody The biological activity of OX40 antibody is lost, showing high stability.
  • the term "stable" antibody formulation is an antibody in a formulation that retains an acceptable degree of physical and / or chemical stability after storage under specific conditions. Although the antibody contained in the antibody preparation does not 100% maintain its chemical structure after storage for a specific time, it is usually maintained at about 90%, 95%, 96%, 97%, 98% after storage for a specific time.
  • an anti-OX40 antibody formulation of the invention substantially retains its physical and chemical stability after storage.
  • the storage time is usually selected based on the expected shelf life of the formulation.
  • the stability test is performed by subjecting the antibody formulation to various stress tests. These tests can indicate the extreme conditions that the formulated antibody preparations may encounter during manufacturing, storage, or transportation, or they can indicate that the instability of the antibodies in the antibody preparations is accelerated under extreme conditions not during manufacturing, storage, or transportation. condition. For example, fill the formulated anti-OX40 antibody preparation into a 5 mL glass bottle for shaking stress, freeze / thaw cycle (ie, freeze-thaw cycle) stress; fill the formulated anti-OX40 antibody preparation into a glass vial to verify that Antibody stability under high temperature stress.
  • freeze / thaw cycle ie, freeze-thaw cycle
  • room temperature refers to a temperature of 15 ° C to 30 ° C, preferably 20 ° C to 27 ° C, and more preferably 25 ° C.
  • high temperature stress refers to the effect on the stability of an antibody formulation after a long storage time at room temperature or even higher temperature (for example, 40 ° C).
  • the formulation After a long storage time, if the formulation is checked for appearance, color and / or clarity, or for example by UV light scattering or size exclusion chromatography, the antibodies in the formulation do not show aggregation, precipitation and / or Cloudiness; or showing very little aggregation, precipitation, and / or cloudiness, then the antibody "maintains its physical stability" in the formulation.
  • the accumulation of antibodies in the formulation can potentially lead to an increased immune response in patients, leading to safety issues. Therefore, there is a need to minimize or prevent aggregation of antibodies in a formulation.
  • the stability of a formulation is measured by determining the percentage of non-aggregated and non-decomposed antibodies in the formulation after storage at a specific temperature for a specific time, where the percentage of non-aggregated and non-decomposed antibodies in the formulation is greater The higher, the more stable the formulation.
  • the percentage of non-aggregated and non-decomposed antibodies can be determined by size exclusion chromatography (eg, size exclusion high performance liquid chromatography).
  • acceptable level of stability means that at least about 92% of non-aggregated and non-decomposed anti-OX40 antibodies are detected in the formulation after storage at a specific temperature for a specific time. In some embodiments, stored at a particular temperature for at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7
  • the acceptable level of stability indicates that at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months or more About 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% of non-aggregated and non-decomposed anti-OX40 antibodies.
  • the specific temperature at which the pharmaceutical formulation is stored can be any temperature from about -80 ° C to about 45 ° C, such as at about -80 ° C, about -30 ° C, about -20 ° C, about 0 ° C, about 4 ° C-8 ° C, about 5 ° C, about 25 ° C, about 35 ° C, about 37 ° C, about 40 ° C, about 42 ° C, or about 45 ° C.
  • the pharmaceutical formulation is considered stable.
  • the antibody after a long storage time, if the chemical structure of the antibody in the formulation is complete, the antibody "maintains its chemical stability" in the formulation. Most chemical instability results from the formation of covalently modified forms of antibodies (eg, charge variants of antibodies).
  • changes in each charge variant component of the anti-OX40 antibody are detected.
  • the charge variant of the anti-OX40 antibody in the antibody formulation is determined by cation exchange high performance liquid chromatography (CEX-HPLC).
  • CEX-HPLC cation exchange high performance liquid chromatography
  • peaks eluting from the CEX-HPLC column earlier than the retention time of the main peak are labeled as "acidic peaks", while those eluting from the CEX-HPLC column later than the retention time of the main peak.
  • the peaks are marked as "basic peaks”.
  • the percentage of acidic components is determined by the ratio of the area of the acidic peak to the sum of the area of the main peak, the acidic peak and the basic peak; the percentage of the main component is the area of the main peak and the main peak, and the acidic peak and basicity.
  • the ratio of the sum of peak areas is determined; the percentage of basic components is determined by the ratio of the area of the basic peak to the sum of the area of the main peak, the acid peak, and the area of the basic peak.
  • deamidation of antibodies can make the antibodies more negatively charged and thus more acidic than non-deamidated antibodies (see, for example, Robinson, N., Protein, Deamidation, PNAS, 2002 (April 16, 1999, 99 (8): 5283-5288).
  • accepted degree of stability means that up to about 25% of the antibody in the formulation is in a more acidic form after storage at a specific temperature for a specific time.
  • stored at a particular temperature for at least 2 weeks, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 Acceptable levels of stability after months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months or more are expressed in Up to about 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody is in an acidic form in the formulation after storage at a specific temperature for a specific time.
  • the temperature at which the pharmaceutical formulation is stored can be any temperature from about -80 ° C to about 45 ° C, such as at about -80 ° C, about -30 ° C, about -20 ° C, about 0 ° C, about 4 ° C. ° C-8 ° C, about 5 ° C, about 25 ° C, or about 45 ° C.
  • the pharmaceutical preparation can be Considered stable.
  • the pharmaceutical preparation It can also be considered stable.
  • a stable anti-OX40 antibody formulation of the invention is administered parenterally to a subject.
  • parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intrasaccular, Intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions.
  • an anti-OX40 antibody formulation of the invention is administered subcutaneously to a subject.
  • a stable anti-OX40 antibody formulation of the invention comprises: (i) an anti-OX40 antibody or antigen-binding fragment thereof that specifically binds to an OX40 molecule; (ii) a buffer; (iii) a stabilizer, and ( iv) surfactant, the pH of the anti-OX40 antibody preparation is about 5.0-8.0.
  • the antibody preparation of the present invention may comprise an anti-OX40 antibody or an antigen-binding fragment thereof that specifically binds to an OX40 molecule.
  • OX40 is known as a costimulatory molecule whose activation can lead to enhanced cell proliferation, survival, effector function, and migration.
  • Anti-OX40 antibodies are disclosed in the prior art as OX40 agonists.
  • WO 2012/027328 discloses the amino acid sequences of the heavy chain and light chain variable regions of anti-OX40 antibody mAb 106-222 and humanized 106-222 (Hu106); anti-OX40 antibody mAb 119-122 and Amino acid sequences of the humanized 119-122 (Hu119) heavy chain and light chain variable regions.
  • U.S. Patent No. 7,959,925, PCT Publication No. WO 2006/121810, and Chinese Patent Application Nos. CN201710185399.9 and CN201710185400.8 also disclose anti-OX40 antibodies as OX40 agonists.
  • the anti-OX40 antibody can activate OX40, thereby inducing the proliferation of effector T lymphocytes, and promoting the immune response against tumor cells expressing tumor-associated antigen (TAA).
  • TAA tumor-associated antigen
  • co-stimulatory molecule refers to a corresponding binding partner on a T cell that specifically binds to a co-stimulatory ligand to mediate a T-cell co-stimulatory response, such as, but not limited to, proliferation.
  • Co-stimulatory molecules are cell surface molecules other than antigen receptors or their ligands that contribute to an effective immune response.
  • the costimulatory molecule is an OX40 molecule.
  • anti-OX40 antibody refers to an antibody capable of binding the OX40 molecule with sufficient affinity so that the antibody can be used as a target A therapeutic and / or prophylactic agent for the OX40 molecule.
  • the degree of binding of an anti-OX40 antibody to an unrelated, non-OX40 protein is less than about 10% of the binding of said antibody to OX40, as measured, for example, by a radioimmunoassay (RIA).
  • the equilibrium dissociation constant (K D ) of the anti-OX40 antibody is ⁇ 1 ⁇ M, ⁇ 100 nM, ⁇ 10 nM, ⁇ 1 nM, ⁇ 0.1 nM, ⁇ 0.01 nM, or ⁇ 0.001 nM.
  • the anti-OX40 antibody is capable of specifically binding OX40 with a high affinity, for example with a K D of 10 -7 M or less, preferably with a K D of 10 -8 M to 10 -12 M, and This mediates a co-stimulatory response.
  • antigen-binding fragments of anti-OX40 antibodies are also included.
  • an "antigen-binding fragment" of an antibody refers to a molecule different from an intact antibody, which contains a portion of the intact antibody and binds to the antigen to which the intact antibody binds.
  • antigen-binding fragments include, but are not limited to, Fv, Fab, Fab ', Fab'-SH, F (ab') 2; diabody; linear antibody; single chain antibody (e.g., scFv); single domain antibody; bivalent or Bispecific antibodies or fragments thereof; camelid antibodies; and bispecific antibodies or multispecific antibodies formed from antigen-binding fragments.
  • epitope refers to a portion of an antigen (eg, OX40) that specifically interacts with an antibody molecule.
  • an antigen eg, OX40
  • CDR region is a sequence that is highly variable in an antibody variable domain and forms a structurally defined loop ("hypervariable loop") and / or contains antigen-contacting residues ( "Antigen contact point”).
  • CDRs are primarily responsible for binding to epitopes.
  • the CDRs of the heavy and light chains are commonly referred to as CDR1, CDR2, and CDR3, and are numbered sequentially from the N-terminus.
  • the CDRs located in the variable domain of the antibody heavy chain are called HCDR1, HCDR2, and HCDR3, while the CDRs located in the variable domain of the antibody light chain are called LCDR1, LCDR2, and LCDR3.
  • each CDR can be determined using any one or a combination of many well-known antibody CDR assignment systems including: For example: Chothia (Chothia et al.
  • the boundaries of the CDRs of the variable regions of the same antibody obtained based on different assignment systems may differ. That is, the CDR sequences of the variable regions of the same antibody defined under different assignment systems are different. Therefore, when it comes to defining an antibody with a specific CDR sequence defined in the present invention, the scope of the antibody also encompasses antibodies whose variable region sequences contain the specific CDR sequences, but due to the application of different protocols (e.g. Different assignment system rules or combinations) resulting in its claimed CDR boundary being different from the specific CDR boundary defined by the present invention.
  • Antibodies with different specificities have different CDRs.
  • CDRs differ from antibody to antibody, only a limited number of amino acid positions within the CDR are directly involved in antigen binding.
  • the minimum binding unit may be a sub-portion of the CDR.
  • the residues of the rest of the CDR sequence can be determined by the structure and protein folding of the antibody. Accordingly, the invention also contemplates any variant of the CDRs given herein. For example, in a variant of a CDR, the amino acid residues of the smallest binding unit may remain unchanged, while the remaining CDR residues as defined by Kabat or Chothia may be replaced by conservative amino acid residues.
  • amino acid residue substitutions are those in which amino acid residues are replaced with amino acid residues having similar side chains.
  • a family of amino acid residues with similar side chains has been defined in the art. These families include those with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), and uncharged polar side chains (e.g., , Glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan, non-polar side chains (e.g., alanine, valine, leucine) , Isoleucine, proline, phenylalanine, methionine), ⁇ -branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., Tyrosine, phenylalanine, tryptophan, histidine).
  • basic side chains
  • one or more amino acid residues of an anti-OX40 antibody can be replaced by other amino acid residues from the same side chain family, and the function of the altered antibody can be tested, especially the same binding characteristics as the OX40 molecule.
  • the change in charge performance on the CDR surface is expected to affect the interface between the antibody and the solvent. Therefore, non-conservative amino acid residue substitutions have an unpredictable effect on maintaining or improving the stability of the antibody in solution.
  • Antibodies or antigen-binding fragments thereof suitable for use in the present invention include, but are not limited to, polyclonal, monoclonal, monovalent, bispecific, heteroconjugate, multispecific, recombinant, heterologous, heterozygous, and chimeric , Humanized (especially CDR-grafted), deimmunized, or human antibodies, Fab fragments, Fab 'fragments, F (ab') 2 fragments, fragments generated from the Fab expression library, Fd, Fv, two Sulfide-linked Fv (dsFv), single chain antibody (e.g. scFv), diabody or tetrabody (Holliger P. et al. (1993) Proc. Natl.
  • Nanobody nanobody
  • anti-idiotypic antibodies including, for example, anti-Id antibodies directed against antibodies of the invention
  • epitope-binding fragments of any of the foregoing.
  • a "human consensus framework” refers to a framework that represents the most frequently occurring amino acid residues in the selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subtype of a variable domain sequence.
  • Antibody in IgG form refers to the IgG form to which the heavy chain constant region of an antibody belongs.
  • the heavy chain constant regions of all antibodies of the same type are the same, and the heavy chain constant regions of different types of antibodies are different.
  • an antibody in the form of IgG1 means that the Ig domain of the heavy chain constant region is the Ig domain of IgG1.
  • Human antibody refers to an antibody having an amino acid sequence that corresponds to the amino acid sequence of an antibody produced by a human or human cell or derived from a non-human source that utilizes a human antibody library or other human Antibody coding sequence. This definition of a human antibody explicitly excludes humanized antibodies that include non-human antigen-binding residues.
  • a “humanized” antibody refers to a chimeric antibody comprising amino acid residues from non-human CDRs and amino acid residues from human FRs.
  • a humanized antibody will comprise substantially all of at least one, usually two variable domains, where all or substantially all CDRs correspond to those of a non-human antibody, and all or substantially all FR corresponds to those of human antibodies.
  • a humanized antibody may optionally comprise at least a portion of a human antibody-derived antibody constant region.
  • a "humanized form" of an antibody (e.g., a non-human antibody) refers to an antibody that has been humanized.
  • isolated antibody is intended to mean an antibody that is substantially free of other antibodies with different antigen specificities, eg, an isolated antibody that specifically binds OX40 is substantially free of antibodies that specifically bind to an antigen other than OX40 .
  • specific binding means that the antibody or antigen-binding fragment thereof forms a complex with the antigen that is relatively stable under physiological conditions. Specific binding is characterized by specifically dissociating OX40 with a dissociation constant of at least about 10 -7 M or less, preferably with K D of 10 -8 M to 10 -12 M, and thereby mediating a co-stimulatory response.
  • Percent (%) amino acid sequence identity with respect to a reference polypeptide sequence is defined as the sequences are aligned (and gaps are introduced if necessary) to obtain the maximum percent sequence identity without regard to any conservative substitutions Following the portion of sequence identity, the percentage of amino acid residues in the candidate sequence to the same amino acid residues in the reference polypeptide sequence. Sequence alignments can be performed using various methods in the art to determine percent amino acid sequence identity, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALIGN (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximum alignment over the full length of the sequences being compared.
  • nucleic acid sequences and protein sequences described herein can be further used as "query sequences" to perform searches against public databases to, for example, identify other family member sequences or related sequences.
  • the amount of the antibody or antigen-binding fragment thereof contained in the antibody preparation of the present invention may vary depending on the specific purpose characteristics of the preparation, the specific environment, and the specific purpose of using the preparation.
  • the antibody formulation is a liquid formulation, which may contain about 1-150 mg / mL, preferably about 10-100 mg / mL, for example, about 15, 20, 25, 30, 35, 40, 45, 50 , 55, 60 mg / mL anti-OX40 antibody or antigen-binding fragment thereof.
  • the invention relates to a formulation having a high concentration of an anti-OX40 antibody, for example, containing 40-150 mg / mL of an anti-OX40 antibody. It is known in the art that such high concentration antibody preparations can be diluted prior to injection, for example if a lower therapeutic antibody concentration is required for a particular therapeutic or prophylactic intervention or when treating smaller weight patients including children.
  • a suitable concentration may be 25 mg / mL or 10 mg / mL.
  • the original formulation can be produced at such a low concentration.
  • the anti-OX40 antibody formulations of the invention described herein in the various embodiments are stable.
  • the anti-OX40 in the antibody formulation of the invention is stored at about -80 ° C, -30 ° C, -20 ° C, 5 ° C, 25 ° C, 37 ° C, 40 ° C, or 45 ° C for 6 months.
  • the purity of the antibody is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more, as determined by size exclusion chromatography.
  • the anti-OX40 in the antibody formulation of the present invention is stored at about -80 ° C, -30 ° C, -20 ° C, 5 ° C, 25 ° C, 37 ° C, 40 ° C, or 45 ° C for 6 months. At least 50% of the antibody is in a non-basic and non-acidic form (ie, a main peak or a major charge form), as determined by cation exchange chromatography.
  • Exemplary anti-OX40 antibodies that can be included in the antibody formulations of the invention are, for example, the anti-OX40 antibodies disclosed in CN201710185399.9 and CN201710185400.8, ADI-20057, ADI-23504, ADI-23507, ADI-23509, ADI-20112, ADI-25650, ADI-25651, ADI-25652, ADI-25653, ADI-25654, ADI-20078, ADI-23515, ADI-23518, ADI-23519, ADI-20048, ADI-20096, ADI-20051, ADI- 20065, ADI-20066, ADI-20118, or ADI-20113, each of which contains the sequences shown in the table below.
  • the anti-OX40 antibody in the antibody preparation of the invention comprises a heavy chain variable region VH and / or a light chain variable region VL, wherein
  • VH shown as SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103 and 104 CDRs of the three complementary determining regions,
  • HCDR1 selected from the amino acid sequences of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 16, and 17 SEQ ID NO: 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36 of the amino acid sequence of HCDR2, and selected from SEQ ID NO: a combination of HCDR3 in the amino acid sequence of 37, 38, 39, 40, 41, 42, 43, and 44; or
  • each of the three CDRs contains at least one and no more than 5, 4, 3, 2 or 1 amino acid changes (preferably amino acid substitutions, preferably conservative substitutions) )the sequence of;
  • LCDR1 selected from the amino acid sequence of SEQ ID NO: 45, 46, 47, 48, 49, and 50
  • LCDR2 selected from the amino acid sequence of SEQ ID NO: 51, 52, 53, 54, 55, and 56
  • a combination of LCDR3 selected from the amino acid sequences of SEQ ID NO: 57, 58, 59, 60, 61, 62, 63, 64, 65 and 66; or
  • each of the three CDRs contains at least one and no more than 5, 4, 3, 2 or 1 amino acid changes (preferably amino acid substitutions, preferably conservative substitutions) )the sequence of.
  • the anti-OX40 antibody in the antibody preparation of the present invention comprises a heavy chain variable region VH and / or a light chain variable region VL, wherein,
  • the heavy chain variable region VH comprises and is selected from SEQ ID NO: 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102 , 103 and 104 have an amino acid sequence of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity or 100% identity Its composition
  • the light chain variable region VL comprises at least 90%, 91%, 92%, and an amino acid sequence selected from the group consisting of SEQ ID NO: 115, 116, 117, 118, 119, 120, 121, 122, 123, and 124, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical or 100% identical amino acid sequences or consist of them.
  • the anti-OX40 antibody in the antibody preparation of the present invention comprises a heavy chain and / or a light chain, wherein the heavy chain comprises and is selected from SEQ ID NO: 125, 126, 127, 128, 129 , 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154 , 155, 156, 157, 158, 159, 160, 161, and 162 have an amino acid sequence of at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% Identity or consist thereof, said light chain comprising at least 90%, 91%, 92% of the amino acid sequence selected from the group consisting of Or consists of an amino acid sequence that is%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%
  • Suitable buffering agents for use in the present invention include, but are not limited to, organic acid salts such as citric acid, ascorbic acid, gluconic acid, succinic acid, tartaric acid, succinic acid, acetic acid or phthalic acid salts; trimethylolaminomethane , Or phosphate buffer, or a combination thereof.
  • an antibody formulation of the invention comprises such a buffer or pH adjuster to provide improved pH control.
  • the liquid formulation of the invention has a pH between 5.0 and 8.0, between 5.0 and 7.0, between 5.5 and 7.0, or between 6.5 and 7.0.
  • an antibody of the invention has a pH of about 5.0, 6.0, 7.0, 8.0.
  • the concentration of the buffering agent contained in the antibody formulation of the present invention is about 0.1-50 mg / ml, preferably about 1-20 mg / ml, for example, about 1.5, 2, 2.5, 3, 3.5, 4 , 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8 mg / ml.
  • Suitable stabilizers used in the present invention can function as, for example, viscosity enhancers, fillers, solubilizers, and the like.
  • the stabilizer may be ionic or non-ionic.
  • the stabilizer may be a non-ionic stabilizer, such as sugar.
  • the sugar as a stabilizer, it includes, but is not limited to, monosaccharides, such as sugar, maltose, galactose, glucose, D-mannose, sorbose, etc .; disaccharides, such as lactose, sucrose, trehalose, cellobiose, etc ; Polysaccharides such as raffinose, melezitose, maltodextrin, dextran, starch, etc .; and sugar alcohols such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucositol) ), Etc., and their combinations.
  • the sugar may be sucrose, trehalose, raffinose, maltose, sorbitol, or mannitol.
  • the sugar is sucrose.
  • this includes salts, for example, NaCl.
  • surfactant refers to an organic substance having an amphiphilic structure; that is, they are composed of groups with opposite solubility tendencies, usually oil-soluble hydrocarbon chains and water-soluble ionic groups group.
  • the surfactant in the liquid formulation of the present invention is a non-ionic surfactant, such as an alkyl poly (ethylene oxide).
  • a non-ionic surfactant such as an alkyl poly (ethylene oxide).
  • Specific non-ionic surfactants that can be included in the formulations of the invention include, for example, polysorbates such as polysorbate-80, polysorbate-60, polysorbate-40, or polysorbate -20; Plonick et al.
  • the amount of the non-ionic surfactant contained in the antibody preparation of the present invention may vary depending on the specific purpose characteristics of the preparation, the specific environment, and the specific purpose of using the preparation.
  • the formulation may contain a concentration of about 0.01-5 mg / ml, preferably about 0.1-2 mg / ml, such as about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0 mg / ml ml of Polysorbate-80 or Plonic.
  • contemplated excipients that can be utilized in the antibody liquid formulations of the present invention include, for example, flavoring agents, antimicrobial agents, sweeteners, antioxidants, antistatic agents, gelatin, and the like.
  • sweeteners for example, peppermints, peppermints, and peppermints
  • antistatic agents for example, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium sulfate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium sulfate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium
  • the antibody formulation according to the invention as described herein in the various embodiments is stable so that even after storage at 25 ° C or 40 ° C for 4 weeks, 1 month or 3 months, anti-OX40 is measured by SEC-HPLC.
  • the purity of the antibody is greater than 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.
  • the antibody preparation of the present invention comprising an anti-OX40 antibody can be used to treat, ameliorate or prevent a variety of diseases or conditions.
  • Anti-OX40 antibody-containing formulations are particularly useful for treating, ameliorating, or preventing cancer, immune-related diseases, and T-cell dysfunction diseases.
  • an antibody formulation of the invention comprising an anti-OX40 antibody can be used to treat, ameliorate or prevent cancer.
  • the cancers include, but are not limited to, B-cell lymphoma (including low / follicular non-Hodgkin's lymphoma (NHL), small lymphocytic (SL) NHL, intermediate / follicular NHL, intermediate diffuse NHL, Advanced Immune Cellular NHL, Advanced Lymphoblastic NHL, Advanced Small Angioblastic NHL, Bulk Disease NHL, Mantle Cell Lymphoma, AIDS-Related Lymphoma, and Waldenstrom's ( Waldenstrom) macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myelogenous leukemia, and post-transplant lymphoproliferative disorder (PTLD) And abnormal vascular proliferation associated with phakomatoses, edema (such as those associated with brain tumors), B cell proliferative disorders, and
  • More specific examples include, but are not limited to, relapsed or refractory NHL, front-line lower NHL, stage III / IV NHL, chemotherapy-resistant NHL, precursor B lymphoblastic leukemia and / or lymphoma, small lymph Cellular lymphoma, B-cell chronic lymphocytic leukemia and / or prelymphocytic leukemia and / or small lymphocytic lymphoma, B-cell prelymphocytic lymphoma, immune cell tumor and / or lymphoplasmic ( lymphoplasmacytic lymphoma, lymphocytoplasmic lymphoma, marginal B-cell lymphoma, marginal spleen lymphoma, extranodal marginal zone-MALT lymphoma, nodal marginal zone lymphoma, Hairy cell leukemia, plasmacytoma and / or plasma cell myeloma, low / follicular lymphoma, intermediate / follicular NHL, mantle cell lymphoma, folli
  • the antibody formulations of the invention are used to treat, ameliorate, or prevent lung cancer (eg, non-small cell lung cancer), liver cancer, gastric cancer, or colon cancer.
  • lung cancer eg, non-small cell lung cancer
  • liver cancer eg, gastric cancer, or colon cancer.
  • the antibody preparation of the present invention can be administered to a subject or patient. Administration is usually by infusion or by syringe. Accordingly, the invention provides a delivery device (e.g., a syringe) that includes an antibody preparation (e.g., a pre-filled syringe) of the invention.
  • a delivery device e.g., a syringe
  • an antibody preparation e.g., a pre-filled syringe
  • the patient will receive an effective amount of an anti-OX40 antibody as the main active ingredient, that is, an amount sufficient to treat, ameliorate or prevent the disease or condition of interest.
  • Therapeutic effects may also include reducing physical symptoms.
  • the optimal effective amount and concentration of antibodies for any particular subject will depend on a number of factors, including the patient's age, weight, health and / or gender, the nature and extent of the disease, the activity of the specific antibody, Its clearance rate, and also includes any possible other treatments administered in combination with the antibody formulation.
  • the effective amount delivered can be determined within the judgment of the clinician.
  • an effective dose may be about 0.005 mg / kg body weight to about 50 mg / kg body weight, or about 0.05 mg / kg body weight to about 10 mg / kg body weight.
  • HERCEPTIN TM administered at an initial loading dose of 4 mg / kg body weight and a weekly maintenance dose of 2 mg / kg body weight
  • RITUXAN TM administered at 375 mg / m 2 weekly
  • SYNAGIS TM is administered intramuscularly at 15 mg / kg body weight
  • the invention also provides a formulation of the invention as described herein in various embodiments for use as a medicament, for example, for delivering an anti-OX40 antibody to a mammal, or for treating, preventing or ameliorating one of the diseases and conditions described above Or more.
  • the mammal is preferably a human, but may also be, for example, a horse or a cow or a dog or a cat.
  • These antibodies are ideally those that match the target species, for example, human anti-OX40 antibodies for humans, horse anti-OX40 antibodies for horses, and dog anti-OX40 antibodies for dogs. If a native host antibody is not available, this can be accomplished by transferring CDR residues (usually, one or more framework residues) from the donor antibody to the recipient framework from the host species. An antibody specifically transfers, for example, to humanization. Equine, bovine, canine and feline antibodies are known in the art. The antibody will bind OX40 of the target species, but it can also cross-react with OX40 from other species.
  • the dose can be a single dose schedule or a multiple dose schedule.
  • CE-SDS sodium alkyl sulfate capillary electrophoresis
  • Sample tray temperature about 10 °C
  • CE-SDS Capillary electrophoresis with sodium lauryl sulfate
  • the antibody preparation sample was diluted with ultrapure water to 10.0 mg / ml, and 10 ⁇ l of the diluted sample was taken, and a pH 6.5 sample buffer solution was sequentially added thereto (the sample buffer solution was obtained by pipetting 200 ⁇ l of pH 6.5 citric acid-phosphate buffer solution).
  • Add 47 ⁇ l 10% SDS add ultrapure water to 1000 ⁇ l to prepare) 85 ⁇ l, 10kDa internal standard (Beckman, US, article number 390953) 2 ⁇ l, 250mmol / L N-ethylmaleimide (NEM) 5 ⁇ l, After thorough mixing, heat at 70 ° C for 10 minutes, and transfer to the sample tube after cooling.
  • CE-SDS analysis was performed using a Beckman PA800 Plus capillary electrophoresis instrument.
  • the sampling voltage was -5kV
  • the sampling time was 20 seconds
  • the separation voltage was -15kV and -16.5kV
  • the analysis time was 35 minutes and 36 minutes, respectively.
  • New anti-OX40 antibodies that specifically bind to OX40 were prepared and purified according to CN201710185399.9 and CN201710185400.8, with the antibody names shown in Table 6 below, and their sequence characteristics are summarized in Tables 1 to 5 above. .
  • Anti-OX40 antibody name ADI-20057 ADI-23504 (offspring of ADI-20057) ADI-23507 (ADI-20057 offspring) ADI-23509 (offspring of ADI-20057) ADI-20112 ADI-25650 (offspring of ADI-20112) ADI-25651 (offspring of ADI-20112) ADI-25652 (offspring of ADI-20112) ADI-25653 (offspring of ADI-20112) ADI-25654 (offspring of ADI-20112)
  • ADI-20078 ADI-23515 (offspring of ADI-20078) ADI-23518 (offspring of ADI-20078) ADI-23519 (offspring of ADI-20078) ADI-20048 ADI-20096 ADI-20051 ADI-20065 ADI-20066 ADI-20118 ADI-20113
  • This example evaluates the effect of the pH of the buffer on the stability of the anti-OX40 antibody.
  • Buffers were prepared at different pH values according to Table 7 below.
  • the concentration of each of the above-mentioned anti-OX40 antibodies in the buffer solution of each pH value was 15 mg / ml. After filtration, aliquot and perform the following measurement.
  • each of the above samples containing 15 mg / ml of anti-OX40 antibody was placed in a 40 ° C ⁇ 2 ° C constant temperature and humidity box, and samples were taken on day 0, day 1, 5 and 10.
  • the appearance of each sample and the presence of visible foreign matter were observed; the protein content in each sample was measured using an ultraviolet visible spectrophotometer (manufactured by Shimadzu, Japan, model UV-1800); and efficient through size exclusion
  • the purity (%) of each sample was measured by liquid chromatography (SEC-HPLC) method; and the charge variant (%) of each sample was measured by cation exchange high performance liquid chromatography (CEX) HPLC. The results are shown below.
  • Table 8 lists the results of determination of protein content in the sample of antibody ADI-20112 (IgG1 type).
  • N / A means this item is not set.
  • the change in sample purity is mainly due to the degradation of antibody proteins after high temperature acceleration; at pH 7.0, the change in sample purity is mainly due to the aggregation of antibody proteins after high temperature acceleration; at pH 8 Under .0 conditions, the initial purity of the antibody protein in the sample was slightly lower, mainly due to the aggregation of the protein.
  • Table 9 lists the results of measuring the protein content of a sample of the antibody ADI-20112 (IgG1 type).
  • the anti-OX40 antibody ADI-20112 was formulated in the buffer at pH 6.0 or pH 7.0, higher purity was observed, indicating that the stability of the anti-OX40 antibody in the pH 6.0 or pH 7.0 buffer was better.
  • each anti-OX40 antibody sample changed at high temperature (40 ⁇ 2 ° C).
  • acid variants of the antibody at pH 8.0, the acid component of the anti-OX40 antibody ADI-20112 (IgG1 type) sample Significantly increased, samples with pH 7.0 followed, samples with pH 5.0 and pH 6.0 had the smallest change; for the main component of the antibody, the main component of the sample with pH 7.0 had the smallest change; for basic variants of the antibody, each The basic components of the sample under pH conditions have a consistent decrease compared to the basic components at day 0.
  • the results are shown in Figures 1-4 of the specification.
  • each of the above samples containing 15 mg / ml of anti-OX40 antibody was shaken at 650 rpm in the dark at 25 ° C, and samples were taken on day 0, day 1, 3, and 5 days.
  • the appearance of each sample and the presence of visible foreign matter were observed; the protein content in each sample was measured using an ultraviolet visible spectrophotometer (manufactured by Shimadzu, Japan, model UV-1800); and by the SEC-HPLC method The purity (%) of each sample was measured.
  • Table 10 lists the protein content measurement results of the antibody ADI-20112 (IgG1 type) sample.
  • N / A means this item is not set.
  • Table 11 lists the purity measurement results of the antibody ADI-20112 (IgG1 type) sample.
  • N / A means this item is not set.
  • each of the above samples containing 15 mg / ml of anti-OX40 antibody was frozen (-30 ° C or lower) and thawed (using a water bath at a temperature of 25 ° C), and samples were taken at the 0th, 3rd, and 6th cycles.
  • the appearance of each sample and the presence of visible foreign matter were observed; the protein content of each sample was measured using an ultraviolet visible spectrophotometer (manufactured by Shimadzu, Japan, model UV-1800); The purity (%) of each sample was measured.
  • Table 12 lists the protein content measurement results of the antibody ADI-20112 (IgG1 type) sample.
  • N / A means this item is not set.
  • Freezing and thawing cycles were performed under conditions of pH of about 5.0, 6.0, 7.0, and 8.0, and the purity of each group of samples was measured. The results showed that the purity of each group of samples was not significantly affected under freeze-thaw conditions.
  • Table 13 lists the purity measurement results of the antibody ADI-20112 (IgG1 type) sample.
  • N / A means this item is not set.
  • liquid preparations with anti-OX40 antibodies were prepared, with a pH of about 7.0.
  • the components of the liquid preparations are shown in Table 14.
  • the previously prepared anti-OX40 antibody sample solution was replaced with the prepared buffer solution of the four preparations, and then the anti-OX40 antibody solution in the sample solution was about 25 mg / ml.
  • the bacteria were filtered and sterilized. Packed in a 15R vial, 4ml / bottle, freeze-dried the sample, tested by high temperature after stoppering and capping, and tested for stability.
  • the lyophilized preparation was placed under conditions of 40 ° C ⁇ 2 ° C and 25 ° C ⁇ 2 ° C, and samples were taken at 0 days, 2 weeks, 4 weeks, and 3 months.
  • the lyophilized preparation sample was reconstituted with sterile water to a volume close to that before lyophilization, and the protein content of the lyophilized preparation after reconstitution was determined to be about 26.0 mg / ml.
  • the four formulations were observed for up to 3 months at a temperature of 40 ° C ⁇ 2 ° C and 25 ° C ⁇ 2 ° C.
  • Formulations 1-4 have an acceptable degree of stability.
  • Formulation 2 was slightly inferior to Formulation 1 in terms of turbidity, purity (SEC-HPLC method) and charge variant (CEX-HPLC method) indicators;
  • Formulation 3 and Formulation 4 were stable for each charge variant (CEX-HPLC method). Slightly lower than Formulation 1.
  • Each stability index of Formulation 1 was better than other formulations.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Medicinal Preparation (AREA)

Abstract

L'invention concerne une formulation contenant un anticorps anti-OX40, et en particulier une formulation médicale d'un anticorps, d'un fragment de liaison à l'antigène, d'un agent tampon, d'un stabilisant et/ou d'un tensioactif qui contient des molécules qui se lient spécifiquement à OX40. L'invention concerne également des utilisations des formulations dans le traitement et la prévention de maladies.
PCT/CN2019/107829 2018-09-25 2019-09-25 Formulation contenant un anticorps anti-ox40, son procédé de préparation et son utilisation WO2020063668A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201980006061.XA CN111683681B (zh) 2018-09-25 2019-09-25 包含抗ox40抗体的制剂、其制备方法及其用途

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201811113627.2 2018-09-25
CN201811113627 2018-09-25

Publications (1)

Publication Number Publication Date
WO2020063668A1 true WO2020063668A1 (fr) 2020-04-02

Family

ID=69951044

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/107829 WO2020063668A1 (fr) 2018-09-25 2019-09-25 Formulation contenant un anticorps anti-ox40, son procédé de préparation et son utilisation

Country Status (3)

Country Link
CN (1) CN111683681B (fr)
TW (1) TWI726426B (fr)
WO (1) WO2020063668A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024078521A1 (fr) * 2022-10-12 2024-04-18 苏州创胜医药集团有限公司 Formulation comprenant un anticorps anti-vegfr2

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104487457A (zh) * 2012-05-30 2015-04-01 中外制药株式会社 靶组织特异性抗原结合分子
WO2016200836A1 (fr) * 2015-06-08 2016-12-15 Genentech, Inc. Méthodes de traitement du cancer au moyen d'anticorps anti-ox40
WO2018017888A1 (fr) * 2016-07-20 2018-01-25 Igm Biosciences, Inc. Molécules multimériques fixant ox40 et leurs utilisations
WO2018032020A1 (fr) * 2016-08-08 2018-02-15 Aeglea Bio Therapeutics, Llc Compositions et méthodes pour le traitement du cancer par déplétion en arginine et à l'aide d'agents d'immuno-oncologie

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104487457A (zh) * 2012-05-30 2015-04-01 中外制药株式会社 靶组织特异性抗原结合分子
WO2016200836A1 (fr) * 2015-06-08 2016-12-15 Genentech, Inc. Méthodes de traitement du cancer au moyen d'anticorps anti-ox40
WO2018017888A1 (fr) * 2016-07-20 2018-01-25 Igm Biosciences, Inc. Molécules multimériques fixant ox40 et leurs utilisations
WO2018032020A1 (fr) * 2016-08-08 2018-02-15 Aeglea Bio Therapeutics, Llc Compositions et méthodes pour le traitement du cancer par déplétion en arginine et à l'aide d'agents d'immuno-oncologie

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024078521A1 (fr) * 2022-10-12 2024-04-18 苏州创胜医药集团有限公司 Formulation comprenant un anticorps anti-vegfr2

Also Published As

Publication number Publication date
CN111683681A (zh) 2020-09-18
TWI726426B (zh) 2021-05-01
TW202033179A (zh) 2020-09-16
CN111683681B (zh) 2023-03-24

Similar Documents

Publication Publication Date Title
JP2023109942A (ja) 単独およびプログラム死受容体１（ｐｄ－１）抗体と組み合わされた抗ｃｔｌａ４抗体の安定な製剤、ならびにその使用方法
JP2024016177A (ja) 単独のおよびプログラム死受容体１（ｐｄ－１）抗体と組み合わされた抗ｔｉｇｉｔ抗体の安定な製剤、ならびにその使用方法
TWI761869B (zh) 包含抗cd47/pd-l1雙特異性抗體的製劑及其製備方法和用途
TWI764097B (zh) 包含抗ｃｄ４７抗體的製劑及其製備方法和用途
TW201347791A (zh) 抗體調配物
WO2021143826A1 (fr) Protéine de mort cellulaire 1 anti-programmée recombinante et préparation d'anticorps bispécifique anti-cluster d'antigène de différenciation 137 et son utilisation
TWI802942B (zh) Pd-l1/lag-3雙特異性抗體製劑及其製備方法和用途
CN114146174B (zh) 抗pd-l1/ox40双特异性抗体制剂及其制备方法和用途
TW202128133A (zh) 重組全人源抗ｔｉｇｉｔ單株抗體製劑及其製備方法和用途
TW202034901A (zh) 抗體配方
WO2020063668A1 (fr) Formulation contenant un anticorps anti-ox40, son procédé de préparation et son utilisation
KR20230009897A (ko) 항-IL-23p19 항체를 포함하는 제제, 이의 제조방법 및 용도
TW202222345A (zh) 包含抗ｔｉｇｉｔ／ｐｄ－１雙特異性抗體的製劑及其製備方法和用途
RU2829156C2 (ru) Композиция, содержащая анти-pd-1/her2 биспецифическое антитело, способ ее получения и ее использование
AU2018263837B2 (en) Stable formulations of anti-CTLA4 antibodies alone and in combination with programmed death receptor 1 (PD-1) antibodies and methods of use thereof
WO2020259593A1 (fr) Préparations contenant un anticorps anti-lag-3, leur procédé de préparation et leur utilisation
WO2024240224A1 (fr) Formulation comprenant un anticorps anti-ccr8 et son utilisation
CN116803420A (zh) 靶向PD-1和TGFβ的双功能蛋白药物组合物及其用途
WO2023134771A1 (fr) Composition pharmaceutique d'anticorps anti-ctla-4 et son utilisation
JP2024528724A (ja) 抗ｐｄ－１抗体医薬組成物及びその使用
CN116710071A (zh) Pd1/pd-l1抗体的皮下施用
TW202114639A (zh) 包含抗ｐｄ－１／ｈｅｒ２雙特異性抗體的製劑及其製備方法和用途
CN112675300A (zh) 包含抗gitr抗体的制剂及其制备方法和用途

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19866457

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19866457

Country of ref document: EP

Kind code of ref document: A1