CN113456582A - Liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody - Google Patents
Liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody Download PDFInfo
- Publication number
- CN113456582A CN113456582A CN202010235536.7A CN202010235536A CN113456582A CN 113456582 A CN113456582 A CN 113456582A CN 202010235536 A CN202010235536 A CN 202010235536A CN 113456582 A CN113456582 A CN 113456582A
- Authority
- CN
- China
- Prior art keywords
- monoclonal antibody
- ser
- liquid formulation
- val
- buffer system
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39591—Stabilisation, fragmentation
-
- 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
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- 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
- A61K47/16—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
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- 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
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- 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
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
Abstract
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a liquid preparation of a recombinant humanized anti-PD-1 monoclonal antibody, which comprises 10mg/ml of a sugarless anti-PD-1 monoclonal antibody, 10-20 mM of a buffering agent, 220-270 mM of a stabilizing agent, 0.02-0.06% (w/v) of a solubilizer, and a buffering system for adjusting the pH to 5.5-6.5. The invention provides a liquid preparation suitable for intravenous administration, which can maintain stability during storage and has low specificity and fragmentation thereof by further optimizing the contents of a buffer solution, a stabilizing agent and a solubilizing agent. And after the preparation is placed for 24 months at the temperature of 2-8 ℃, the appearance, concentration and pH, antibody purity and charge heteroplasmon of the preparation have no obvious change.
Description
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a liquid preparation of a recombinant humanized anti-PD-1 monoclonal antibody.
Background
PD-1 is an immunosuppressive receptor, mainly in activated T cells and B cells, as a T cell inhibitory receptor, and can inhibit the activity of T lymphocytes and related in vivo cellular immune response by combining with ligand PD-L1, can limit the function of T cell effectors in tumor cells, and has an important role in tumor immune escape. The anti-PD-1 monoclonal antibody can block Programmed Death (PD) -1 receptors on the surfaces of activated T cells, can save exhausted T cells by inhibiting a PD-1 and PD-1 ligand (PD-L1) passage, enhances the anti-tumor immunity, and has good targeting potential in the aspect of treating tumors. In response to this advantage, the development of anti-PD-1 monoclonal antibodies such as opdivo, keytruda and other monoclonal antibodies, acetonitrile, was approved for the market abroad, but these antibodies have a low response rate as a single drug. The glycosylation modification of the monoclonal antibody is knocked out, although the process development is stable, the uniformity is high, and a better clinical effect is expected to be obtained, compared with other anti-PD-1 monoclonal antibodies, the sensitivity of the monoclonal antibody to trypsin and the like is obviously increased due to the removal of glycosylation, the structural flexibility is enhanced, the monoclonal antibody has a tendency of polymer formation under the condition of low pH, and the aggregation and the precipitation are easy. In order to prevent the protein from denaturation during storage before use, loss of activity or structural integrity due to aggregation, formulation processes for this aglycosylated anti-PD-1 monoclonal antibody are necessary.
Patent CN103429264A discloses a lyophilized formulation of an anti-human PD-1 antibody or an antigen-binding fragment thereof, comprising 25mg/ml of the anti-human PD-1 antibody or an antigen-binding fragment thereof, 10mM histidine buffer at pH 5.0-6.0, 70mg/ml of sucrose, 0.02mg/ml of polysorbate 80.
The patent CN107334728A discloses a solution preparation of an anti-human PD-1 monoclonal antibody, which comprises 100mg/ml of the anti-human PD-1 monoclonal antibody, 20mM of histidine buffer, 250mM of sucrose, 6-8 mg/ml of poloxamer 188, 20-100 mg/ml of sulfobutyl ether beta cyclodextrin and 2-8 mg/ml of benzyl alcohol.
Patent CN107325180A discloses a monoclonal antibody preparation against human PD-1 suitable for subcutaneous injection, which comprises 100mg/ml of monoclonal antibody against human PD-1, 10mM of phosphate buffer, 10mM of citrate buffer, 250mM of mannitol, 20mM of aspartic acid, and 8mg/ml of Tween 20.
Patent CN106390115A discloses a stable preparation of humanized monoclonal antibody, which comprises 25-50 mg/ml, pH6.0, 10-20 mM citric acid buffer solution, 150mM mannitol and 50mM sodium chloride, and 0.20% polysorbate 80.
CN107198773A discloses a liquid preparation of a recombinant anti-PD-L1 fully human monoclonal antibody, which comprises 30mg/ml of the recombinant anti-PD-L1 fully human monoclonal antibody, 20mM histidine-histidine hydrochloride, 150mM mannitol, 55mM sodium chloride and 0.01 wt% of polysorbate 80.
CN105793288A discloses an aqueous pharmaceutical formulation of PDL1 monoclonal antibody, comprising 60mg/ml anti-PDL 1 monoclonal antibody, 20mM histidine acetate or sodium acetate buffer, 120mM sucrose, ph 5.8.
In the above patents, the stability of the antibody preparation is improved by screening auxiliary materials and adding a stabilizer or an additive, and the increase of the auxiliary materials can bring potential safety hazards to clinical medication. In addition, different PD-1 antibodies differ in structure and performance properties, and therefore the formulation of the prior art is not suitable for use with aglycosylated anti-PD-1 monoclonal antibodies.
CN110354073A discloses a liquid formulation of an immunosuppressant monoclonal antibody, comprising 25mg/ml aglycosylated anti-PD-1 monoclonal antibody, 15mM histidine at pH6.0, 270mM sucrose and 0.4mg/ml polysorbate 20. The high-temperature-resistant and high-temperature-resistant composite material is relatively stable when placed at 2-8 ℃ for 6 months, but the stability of the high-temperature-resistant and high-temperature-resistant composite material is still to be further improved. There is therefore also a need to provide a more stable liquid formulation without glycosylation of anti-PD-1 monoclonal antibodies.
Disclosure of Invention
Based on the defects of the prior art, the invention aims to provide a stable liquid preparation of the recombinant humanized anti-PD-1 monoclonal antibody.
The technical scheme of the invention is as follows:
a liquid preparation of a recombinant humanized anti-PD-1 monoclonal antibody comprises 10mg/ml of a sugarless anti-PD-1 monoclonal antibody, 10-20 mM of a buffering agent, 220-270 mM of a stabilizing agent, 0.02-0.06% of a solubilizer, and the pH of a buffering system is adjusted to 5.5-6.5.
Preferably, the buffer is histidine hydrochloride or citric acid/sodium citrate, more preferably histidine hydrochloride.
Preferably, the stabilizer is sucrose or mannitol; further preferred is sucrose.
Preferably, the solubilizer is polysorbate 80 or polysorbate 20.
In one embodiment, a liquid formulation of a recombinant humanized anti-PD-1 monoclonal antibody comprises 10mg/ml of a sugarless anti-PD-1 monoclonal antibody, 10-20 mM histidine hydrochloride buffer solution, 220-250 mM sucrose, 0.02-0.06% (w/v) polysorbate 80, and a buffer system for adjusting pH to 6.0-6.5.
Preferably, the concentration of the histidine-hydrochloric acid buffer solution is 10 mM.
Preferably, the sucrose concentration is 220 mM.
Preferably, the polysorbate 80 content is 0.02% (w/v).
Preferably, the pH is 6.0.
In a preferred protocol, a liquid formulation of a recombinant humanized anti-PD-1 monoclonal antibody comprises the following components:
the invention also provides a preparation method of the liquid preparation of the recombinant humanized anti-PD-1 monoclonal antibody, which comprises the following steps: weighing a buffering agent with a prescription amount, adding water for injection to dissolve, adjusting pH, performing ultrafiltration concentration by using the prepared buffer system aglycosylation anti-PD-1 monoclonal antibody protein stock solution, replacing an original buffer system of aglycosylation anti-PD-1 monoclonal antibody protein, adding sucrose and polysorbate 80 with the prescription amount, stirring uniformly, adjusting the protein concentration to 10mg/ml by using the buffer system, performing sterile filtration by using a 0.22 mu m filter membrane, and filling to obtain the product.
The aglycosylated anti-PD-1 monoclonal antibody of the present invention can be prepared with reference to CN106519034A, and the sequences of the heavy chain and the light chain of the aglycosylated anti-PD-1 monoclonal antibody are as follows:
heavy chain region
MEWSWVFLFFLSVTTGVHSEVQLVESGGGLVKPGGSLRLSCAASGFTFSSYGMSWVRQTPEKRLEWVATISGGGRDTYYPDSVKGRFTISRDNAKNNLYLQMSSLRSEDTALYYCARQKDTSWFVHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK
Light chain region
MSVPTQVLGLLLLWLTDARCEIVLTQSPATLAVSPGERATISCRASESVDDYGISFMNWFQQKPGQPPKLLIYVASNQGSGVPARFSGSGSGTDFTLNIHPMEEDDTAMYFCQQSKEVPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
The invention provides a liquid preparation suitable for intravenous administration, which can maintain stability during storage, and has low polymer and fragmentation thereof by further optimizing the contents of a buffer solution, a stabilizing agent and a solubilizing agent. And after the preparation is placed for 24 months at the temperature of 2-8 ℃, the appearance, concentration and pH, antibody purity and charge heteroplasmon of the preparation have no obvious change. After accelerating for 6 months, the appearance, concentration, activity, pH value, purity (SEC-HPLC) and charge heteroplasmon (CEX-HPLC) of the preparation have no obvious change.
Detailed Description
The invention is further illustrated by the following examples, which are intended to be part of a formulation screening assay and are not intended to limit the scope of the invention. The test methods under specific conditions not specified in the following examples were selected in accordance with conventional methods and conditions, or in accordance with commercial instructions. In the examples, the stability test and the related biological test were carried out according to the specifications of the Chinese pharmacopoeia. The actual substances described in the examples are all pharmaceutical grades, which are sold in the market. The aglycosylated anti-PD-1 monoclonal antibody can be prepared by the method referred to CN 106519034A.
Example 1
The formula is as follows:
the preparation method comprises the following steps:
weighing a buffering agent with a prescription amount, adding water for injection to dissolve, adjusting pH, performing ultrafiltration concentration by using the prepared buffer system aglycosylation anti-PD-1 monoclonal antibody protein stock solution, replacing an original buffer system of aglycosylation anti-PD-1 monoclonal antibody protein, adding sucrose and polysorbate 80 with the prescription amount, stirring uniformly, adjusting the protein concentration to 10mg/ml by using the buffer system, performing sterile filtration by using a 0.22 mu m filter membrane, and filling to obtain the product.
Example 2
The formula is as follows:
the preparation is as in example 1.
Example 3
The formula is as follows:
the preparation is as in example 1.
Example 4
The formula is as follows:
the preparation is as in example 1.
Example 5
The formula is as follows:
the preparation is as in example 1.
Example 6
The formula is as follows:
the preparation method comprises the following steps:
weighing a buffering agent with a prescription amount, adding water for injection to dissolve, adjusting pH, performing ultrafiltration concentration by using the prepared buffer system aglycosylation anti-PD-1 monoclonal antibody protein stock solution, replacing an original buffer system of aglycosylation anti-PD-1 monoclonal antibody protein, adding mannitol and polysorbate 80 with a prescription amount, stirring uniformly, adjusting the protein concentration to 10mg/ml by using the buffer system, performing sterile filtration by using a 0.22 mu m filter membrane, and filling to obtain the product.
Example 7
The formula is as follows:
the preparation method comprises the following steps:
weighing a buffering agent with a prescription amount, adding water for injection to dissolve, adjusting pH, performing ultrafiltration concentration by using the prepared buffer system aglycosylation anti-PD-1 monoclonal antibody protein stock solution, replacing an original buffer system of aglycosylation anti-PD-1 monoclonal antibody protein, adding sucrose and polysorbate 80 with the prescription amount, stirring uniformly, adjusting the protein concentration to 10mg/ml by using the buffer system, performing sterile filtration by using a 0.22 mu m filter membrane, and filling to obtain the product.
Example 8
The formula is as follows:
the preparation method is the same as example 1.
Example 9
The formula is as follows:
the preparation method is the same as example 1.
Verification examples
Three batches of samples were prepared according to examples 1-9, respectively, and the storage stability was investigated using accelerated stability testing and long term testing.
The long-term test is carried out at the temperature of 2-8 ℃, and the test is carried out according to the stability key investigation items at the end of 0 month, 3 months, 6 months, 12 months and 24 months respectively; the accelerated test is carried out at 25 +/-2 ℃ for 6 months, the used equipment can control the temperature to +/-2 ℃, the actual temperature is detected, samples are taken once at the end of 0 month, 3 months and 6 months during the test period, and the test is carried out according to the key stability investigation items. The results are shown in tables 1 and 2.
TABLE 12-8 ℃ long-term stability test results
TABLE 225. + -. 2 ℃ accelerated stability test results
SEQUENCE LISTING
<110> Lunan pharmaceutical group, Inc
<120> liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody
<130> 2020
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 467
<212> PRT
<213> Artificial sequence
<400> 1
Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly
1 5 10 15
Val His Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys
20 25 30
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
35 40 45
Ser Ser Tyr Gly Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu
50 55 60
Glu Trp Val Ala Thr Ile Ser Gly Gly Gly Arg Asp Thr Tyr Tyr Pro
65 70 75 80
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn
85 90 95
Asn Leu Tyr Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Leu
100 105 110
Tyr Tyr Cys Ala Arg Gln Lys Asp Thr Ser Trp Phe Val His Trp Gly
115 120 125
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
130 135 140
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
145 150 155 160
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
165 170 175
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
180 185 190
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
195 200 205
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
210 215 220
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
225 230 235 240
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
245 250 255
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Gln Leu Met
260 265 270
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
275 280 285
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
290 295 300
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr
305 310 315 320
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
325 330 335
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
340 345 350
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
355 360 365
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
370 375 380
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
385 390 395 400
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
405 410 415
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
420 425 430
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Leu
435 440 445
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
450 455 460
Pro Gly Lys
465
<210> 2
<211> 238
<212> PRT
<213> Artificial sequence
<400> 2
Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr
1 5 10 15
Asp Ala Arg Cys Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ala
20 25 30
Val Ser Pro Gly Glu Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser
35 40 45
Val Asp Asp Tyr Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro
50 55 60
Gly Gln Pro Pro Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser
65 70 75 80
Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Asn Ile His Pro Met Glu Glu Asp Asp Thr Ala Met Tyr Phe Cys
100 105 110
Gln Gln Ser Lys Glu Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu
115 120 125
Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
130 135 140
Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu
145 150 155 160
Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175
Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser
180 185 190
Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala
195 200 205
Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
210 215 220
Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
Claims (10)
1. A liquid preparation of a recombinant humanized anti-PD-1 monoclonal antibody is characterized by comprising 10mg/ml of a sugarless anti-PD-1 monoclonal antibody, 10-20 mM of a buffering agent, 220-270 mM of a stabilizing agent, 0.02-0.06% of a solubilizer and a buffer system for adjusting the pH to 5.5-6.5.
2. The liquid formulation of claim 1, wherein the buffer is histidine hydrochloride or citric acid/sodium citrate.
3. The liquid formulation of claim 2, wherein the buffering agent is histidine hydrochloride.
4. The liquid formulation of claim 1, wherein the stabilizer is sucrose or mannitol.
5. The liquid formulation of claim 4, wherein the stabilizer is sucrose.
6. The liquid formulation according to claim 1, wherein the solubilizer is polysorbate 80 or polysorbate 20.
7. The liquid formulation according to any one of claims 1 to 6, comprising the following components in amounts:
8. the liquid formulation of claim 1, wherein the aglycosylated anti-PD-1 monoclonal antibody has the amino acid sequence as follows:
heavy chain region
MEWSWVFLFFLSVTTGVHSEVQLVESGGGLVKPGGSLRLSCAASGFTFSSYGMSWVRQTPEKRLEWVATISGGGRDTYYPDSVKGRFTISRDNAKNNLYLQMSSLRSEDTALYYCARQKDTSWFVHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK；
Light chain region
MSVPTQVLGLLLLWLTDARCEIVLTQSPATLAVSPGERATISCRASESVDDYGISFMNWFQQKPGQPPKLLIYVASNQGSGVPARFSGSGSGTDFTLNIHPMEEDDTAMYFCQQSKEVPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC。
9. A method for preparing the liquid formulation of claim 7, comprising the steps of: weighing a buffering agent with a prescription amount, adding water for injection to dissolve, adjusting pH, performing ultrafiltration concentration by using the prepared buffer system aglycosylation anti-PD-1 monoclonal antibody protein stock solution, replacing an original buffer system of aglycosylation anti-PD-1 monoclonal antibody protein, adding sucrose and polysorbate 80 with the prescription amount, stirring uniformly, adjusting the protein concentration to 10mg/ml by using the buffer system, performing sterile filtration by using a 0.22 mu m filter membrane, and filling to obtain the product.
10. A method for preparing a liquid formulation according to any one of claims 1 to 6 or 8, comprising the steps of: weighing a buffering agent with a prescription amount, adding water for injection to dissolve, adjusting pH, performing ultrafiltration concentration by using the prepared buffer system aglycosylation anti-PD-1 monoclonal antibody protein stock solution, replacing an original buffer system of aglycosylation anti-PD-1 monoclonal antibody protein, adding sucrose and polysorbate 80 with the prescription amount, stirring uniformly, adjusting the protein concentration to 10mg/ml by using the buffer system, performing sterile filtration by using a 0.22 mu m filter membrane, and filling to obtain the product.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010235536.7A CN113456582A (en) | 2020-03-30 | 2020-03-30 | Liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody |
| PCT/CN2020/101516 WO2021196443A1 (en) | 2020-03-30 | 2020-07-11 | Liquid preparation of recombinant humanized anti-pd-1 monoclonal antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010235536.7A CN113456582A (en) | 2020-03-30 | 2020-03-30 | Liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113456582A true CN113456582A (en) | 2021-10-01 |
Family
ID=77864792
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010235536.7A Pending CN113456582A (en) | 2020-03-30 | 2020-03-30 | Liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN113456582A (en) |
| WO (1) | WO2021196443A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114099672A (en) * | 2022-01-28 | 2022-03-01 | 嘉和生物药业有限公司 | Pharmaceutical composition of anti-RANKL humanized monoclonal antibody with enhanced stability |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117427157A (en) * | 2023-10-30 | 2024-01-23 | 广州誉衡生物科技有限公司 | Stable preparation of fully human anti-PD-1 monoclonal antibody |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106519034A (en) * | 2016-12-22 | 2017-03-22 | 安源医药科技（上海）有限公司 | Anti-PD-1 (Programmed Death-1) antibody and application thereof |
| CN107198773A (en) * | 2017-06-08 | 2017-09-26 | 上海药明生物技术有限公司 | Recombinate the liquid preparation of anti-PD L1 human monoclonal antibodies |
| CN110354073A (en) * | 2018-04-09 | 2019-10-22 | 鲁南制药集团股份有限公司 | A kind of liquid preparation of immunosuppressor monoclonal antibody |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2011126338A (en) * | 2008-11-28 | 2013-01-10 | Эбботт Лэборетриз | STABLE COMPOSITIONS OF ANTIBODIES AND METHODS FOR THEIR STABILIZATION |
| WO2012135408A1 (en) * | 2011-03-31 | 2012-10-04 | Merck Sharp & Dohme Corp. | Stable formulations of antibodies to human programmed death receptor pd-1 and related treatments |
| EP3240571A4 (en) * | 2014-12-31 | 2018-06-13 | NovelMed Therapeutics, Inc. | Formulation of aglycosylated therapeutic antibodies |
| CN107325180A (en) * | 2016-04-28 | 2017-11-07 | 上海抗体药物国家工程研究中心有限公司 | A kind of monoclonal antibody formulation suitable for hypodermic anti-human PD-1 |
| EP3518891A1 (en) * | 2016-09-27 | 2019-08-07 | Fresenius Kabi Deutschland GmbH | Liquid pharmaceutical composition |
-
2020
- 2020-03-30 CN CN202010235536.7A patent/CN113456582A/en active Pending
- 2020-07-11 WO PCT/CN2020/101516 patent/WO2021196443A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106519034A (en) * | 2016-12-22 | 2017-03-22 | 安源医药科技（上海）有限公司 | Anti-PD-1 (Programmed Death-1) antibody and application thereof |
| CN107198773A (en) * | 2017-06-08 | 2017-09-26 | 上海药明生物技术有限公司 | Recombinate the liquid preparation of anti-PD L1 human monoclonal antibodies |
| CN110354073A (en) * | 2018-04-09 | 2019-10-22 | 鲁南制药集团股份有限公司 | A kind of liquid preparation of immunosuppressor monoclonal antibody |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114099672A (en) * | 2022-01-28 | 2022-03-01 | 嘉和生物药业有限公司 | Pharmaceutical composition of anti-RANKL humanized monoclonal antibody with enhanced stability |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2021196443A1 (en) | 2021-10-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111971062B (en) | Monoclonal antibody preparation against human PD-1, combined medicament and application thereof | |
| CN111246886B (en) | Pharmaceutical composition of anti-PD-L1 humanized monoclonal antibody | |
| CN108367072A (en) | Aqueous pharmaceutical preparations containing anti-PD-1 antibody As VELUMAB | |
| CN110354073B (en) | Liquid preparation of immunosuppressant monoclonal antibody | |
| EP3563868A1 (en) | Pharmaceutical preparation stably comprising cd147 monoclonal antibody | |
| EP3991745A1 (en) | Formulations containing anti-cd47/pd-l1 bispecific antibody and preparation method therefor and use thereof | |
| TWI802942B (en) | PD-L1/LAG-3 bispecific antibody preparation and its preparation method and use | |
| US20210188996A1 (en) | Formulations of anti-cd38 antibodies for subcutaneous administration | |
| CN113456582A (en) | Liquid preparation of recombinant humanized anti-PD-1 monoclonal antibody | |
| JP2020509065A (en) | Aqueous anti-PD-L1 antibody preparation | |
| EP3811979B1 (en) | Anti-her2 antibody drug conjugate pharmaceutical preparation | |
| CN112566634A (en) | Treatment of stage III NSCLC and remission of treatment-related pathological conditions | |
| TW201907951A (en) | Prolactin receptor antibody for male and female hair loss | |
| WO2021115321A1 (en) | Pharmaceutical taci-fc fusion protein formulation | |
| CN114146174A (en) | anti-PD-L1/OX 40 bispecific antibody preparation and preparation method and application thereof | |
| CN114984207B (en) | anti-PD-1 nano antibody preparation | |
| WO2021147854A1 (en) | Recombinant fully human anti-tigit monoclonal antibody preparations, preparation method therefor and use thereof | |
| CN116261448A (en) | Liquid formulations comprising high concentrations of humanized antibodies for the treatment of IL-6 related diseases | |
| CN115581766A (en) | PD-L1/TGF beta combined bifunctional antibody fusion protein liquid preparation | |
| CN115768799A (en) | anti-HER 2 monoclonal antibody freeze-dried preparation and preparation method thereof | |
| CN113797330A (en) | anti-PD-1 monoclonal antibody liquid preparation | |
| CN113082203B (en) | Liquid preparation of immunosuppressant monoclonal antibody | |
| CN114007648B (en) | Formulations comprising anti-LAG-3 antibodies, methods of making and uses thereof | |
| EP4070817A1 (en) | Liquid preparation containing anti-il-17 antibody | |
| EP4011390A1 (en) | Preparation comprising anti-pd-1/her2 bispecific antibody, and preparation method therefor and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |