CN113521018B - Freeze-dried composition containing anti-HER 2-drug conjugate, freeze-dried preparation, preparation method and application thereof - Google Patents
Freeze-dried composition containing anti-HER 2-drug conjugate, freeze-dried preparation, preparation method and application thereof Download PDFInfo
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- 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
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
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- 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
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- 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
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- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
- A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
- A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
- A61K47/6817—Toxins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention provides a freeze-dried composition containing an anti-HER 2-drug conjugate, which comprises ARX788 and one or more of pharmaceutically acceptable freeze-drying protective agents, buffer salts and surfactants, wherein the pH value of the freeze-dried composition is 5.7-6.3. The invention also provides a freeze-dried preparation containing the HER 2-resisting drug conjugate and a preparation method thereof. The invention also provides the application of the freeze-dried composition and the freeze-dried preparation. The freeze-dried composition and the freeze-dried preparation provided by the invention have the advantages of excellent stability, excellent appearance, short redissolution time, low moisture content, safe and reliable product quality and high production efficiency.
Description
Technical Field
The invention relates to the technical field of biological pharmacy, in particular to a freeze-dried composition containing an anti-HER 2-drug conjugate, a freeze-dried preparation prepared from the freeze-dried composition, and a preparation method and application of the freeze-dried preparation.
Background
Human epidermal growth factor receptor 2 (HER 2/ErbB 2) is one of the epithelial growth factor receptor family members, a transmembrane tyrosine kinase. Amplification of the HER2 gene is present in about 30% of primary human breast cancers, and is sometimes found in a variety of other solid tumors as well. Amplification of the HER2 gene typically results in overexpression such that as many as one million molecules of HER2 protein are present on the surface of each cancer cell. Elevated HER2 protein expression and HER2 gene amplification are associated with poor clinical prognosis, including shortening of recurrence-free survival and Overall Survival (OS) for breast and gastric cancers, and shortening of overall survival for lung, ovarian, colon and pancreatic cancers. Several HER 2-targeted therapeutics, antibodies and antibody-drug conjugates (ADCs, including T-DM 1) have been demonstrated to have clinical anti-cancer activity.
The medicine must be marketed in the form of final preparation, and dosage form selection, the types and the dosage of auxiliary materials, preparation process and the like all have influence on the stability of the product. The qualified product has stable and controllable quality in the period of validity, and is convenient to transport, store and use clinically. The stability of the medicine is one of important indexes for evaluating the effectiveness and safety of the medicine, the stability research is a main basis for determining the storage condition and the service life of the medicine, and scientific basis can be provided for the production, packaging, transportation condition and the like of the medicine. In the case of pharmaceutical products, particularly biological products, the retention of the molecular configuration and biological activity of the active ingredient depends on various covalent and non-covalent forces, and the maintenance of these forces is closely related to the microenvironment in which they are located, such as temperature, light, ion concentration, and mechanical shear force.
ADCs are highly effective and specific agents for the treatment of cancer and other conditions in which the antibody moiety specifically binds to its antigen on the target cell, such that the drug may exert its cytotoxic or other therapeutic effect on the target cell. However, like other protein drugs, antibodies are susceptible to degradation such as oxidation, deamidation, and fragmentation as well as formation of particulates and aggregates. The purpose of the prescription development of the ADC preparation is to ensure that the administration of the ADC preparation is stable and the quality is reliable. However, compared with unconjugated antibody, ADC has a more complex structure and more unstable factors, and the safety, effectiveness and quality controllability of the product can be influenced by the characteristics of a plurality of preparations expressed in the development process of biological preparations. Thus, ADCs present challenges for formulation for therapeutic purposes. To provide ADC drugs that are stable during transport and storage, the choice of carriers, excipients, and/or stabilizers in the pharmaceutical formulation must be carefully selected.
The ADC quality attribute comprises a free small molecule drug aspect, a naked antibody aspect and an antibody drug conjugate aspect. The safety of ADC is influenced by the content of free small molecule drugs, and the naked antibody has inherent key mass attributes of the antibody, such as chemical instability of polymers, fragments, charge heterogeneity, degradation, oxidation and the like, and all the attributes can influence various clinical curative effects such as biological activity, immunogenicity, drug effect, safety and the like. The specific quality attributes of ADC, such as chemical stability of conjugate drugs, drug-antibody ratio, etc., may affect the potency, drug metabolism and safety of ADC.
Disclosure of Invention
Based on the particularity of the ADC formulation, it is an object of the present invention to provide a novel, stable ADC formulation, a lyophilized composition comprising an anti-HER 2-drug conjugate.
It is another object of the present invention to provide a lyophilized formulation comprising an anti-HER 2-drug conjugate and a method for preparing the same.
It is also an object of the present invention to provide the use of the lyophilized composition and the lyophilized formulation.
The lyophilized composition containing the anti-HER 2-drug conjugate provided by the invention comprises ARX788 and one or more of pharmaceutically acceptable lyoprotectants, buffer salts and surfactants, wherein the pH value of the lyophilized composition is 5.7-6.3.
In some preferred embodiments, the pH of the lyophilized composition may be 6.0 to 6.3, and in some most preferred embodiments, the pH of the lyophilized composition may be 6.0.
The ARX788 is an anti-HER 2-ADC, also called anti-HER 2 monoclonal antibody-AS 269 conjugate (structural formula shown in figure 1). The anti-HER 2 antibody is obtained by mutating the 121 th amino acid of the heavy chain of trastuzumab to an acetylphenylalanine residue by using a codon expansion technology, carrying out oximation reaction on the mutated heavy chain and small molecular toxin containing hydroxylamine groups, and carrying out site-specific coupling to obtain a compound antibody with the main drug-to-antibody ratio of 1: 2. This is described in detail in chinese patent CN 201280036296.1.
The pharmaceutically acceptable lyoprotectants, buffer salts and surfactants can be any kind of common cryoprotectants in the pharmaceutical field, especially the kind of common cryoprotectants in ADC preparations. In some preferred embodiments, the buffer salts include, but are not limited to, acetate, phosphate, citrate, histidine, succinate, 2- (N-morpholinyl) ethanesulfonic acid, ethylenediaminetetraacetic acid disodium salt, and the like; such surfactants include, but are not limited to, polysorbate 20, polysorbate 80, and the like; the lyoprotectant includes, but is not limited to, sucrose, trehalose, and the like.
In some preferred embodiments, the lyophilized composition of the present invention comprises 10 to 30mg/mL of ARX788, 5 to 10mM of histidine, 2.5 to 8wt.% of trehalose, and 0.01 to 0.05wt.% of polysorbate 80.
In some more preferred embodiments, the composition of the invention comprises 20mg/mL of ARX788, 5mM histidine, 6wt.% trehalose, and 0.02wt.% polysorbate 80, at a pH of 6.0.
The invention also provides a freeze-dried preparation containing the anti-HER 2-drug conjugate, which is prepared from the freeze-dried composition in any one of the technical schemes.
The invention also provides a preparation method of the freeze-dried preparation, which comprises the following steps:
s1: pre-freezing the freeze-dried composition of any one of the technical schemes for 3 to 6 hours at a temperature of between 50 ℃ below zero and 40 ℃ below zero;
s2: heating the freeze-dried composition treated in the step S1 to-25 to-20 ℃, and keeping the temperature and the vacuum degree of 10-15 Pa for 30-50 h; and
s3: and (3) heating the freeze-dried composition processed in the step (S2) to 25-30 ℃, and keeping the temperature and the vacuum degree of 10-15 Pa for 5-10 h.
In some preferred embodiments, the present invention provides a preparation method, wherein the step S1 comprises: the freeze-dried composition in any one of the technical schemes is cooled to 2-5 ℃ within 0.5-1.5 h, the temperature is kept for 0.5-1.5 h, then the temperature is cooled to-50-40 ℃ within 3-5 h, and the temperature is kept for 3-6 h. In some more preferred embodiments, the step S1 comprises: the freeze-dried composition in any of the technical schemes is cooled to 4 ℃ within 1h, kept warm for 1h, then cooled to-45 ℃ within 3.5h, and kept warm for 5h.
In some preferred embodiments, the preparation method provided by the present invention, the step S2 includes: and (3) heating the freeze-dried composition treated in the step (S1) to-23 to-20 ℃ within 0.2 to 0.8h, and keeping the temperature and the vacuum degree of 12 to 14Pa for 35 to 45h. In some more preferred embodiments, the step S2 includes: and (3) heating the freeze-dried composition treated in the step S1 to-23-20 ℃ within 0.5h, and keeping the temperature and the vacuum degree of 13Pa for 40h.
In some preferred embodiments, the present invention provides a preparation method, wherein the step S3 comprises: and (3) heating the freeze-dried composition treated in the step (S2) to 25-30 ℃ within 2-8 h, and keeping the temperature and the vacuum degree of 12-14 Pa for 5-10 h. In some more preferred embodiments, the step S3 comprises: and (3) heating the freeze-dried composition treated in the step (S2) to 25-30 ℃ within 5h, and keeping the temperature and the vacuum degree of 13Pa for 8h.
The invention also provides the use of the lyophilized composition of any of the above claims or the lyophilized formulation of any of the above claims in the preparation of a medicament for the treatment of cancer.
In the use of the present invention, the cancer includes, but is not limited to, breast cancer, stomach cancer, lung cancer, ovarian cancer, colorectal cancer, urothelial cancer, biliary tract cancer, liver cancer, brain cancer, esophageal cancer, endometrial cancer, uterine cancer, kidney cancer, bladder cancer, thyroid cancer, salivary gland cancer, or pancreatic cancer.
The freeze-dried composition and the freeze-dried preparation provided by the invention are designed particularly for a novel ADC drug ARX788, and through the specific pH value, the composition formula and even the arrangement of the freeze-drying process, the falling of small molecule drugs and the reduction of a main peak of CEX can be effectively reduced, the obtained freeze-dried product has very excellent stability, can keep good stability and biological activity for a long time even at high temperature (such as 40 ℃), and has the advantages of excellent appearance, short re-dissolving time (within one minute and even within 30 s), low moisture content, safe and reliable product quality and high production efficiency. The freeze-dried composition and the freeze-dried preparation provided by the invention have the advantages of simple preparation method and low cost, can be used for treating various cancers, and have wide application prospects.
Drawings
FIG. 1 is a schematic diagram of the structure of ADC drug ARX788, wherein one antibody protein is coupled with two small molecule compounds.
FIGS. 2A and 2B are a CEX trend graph and an activity trend graph, respectively, under the high temperature condition in example 5.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the following specific examples.
The ADC drugs ARX788 and ADC Reference (RS) used in the examples are from New code, zhejiang, bio-medicine, inc., and other reagents or raw materials are commercially available unless otherwise specified.
The detection items and detection conditions used in the examples are as follows:
appearance: visual inspection was performed with a clarity detector.
Protein concentration: cary 100 UV-Vis Spectrophotometer detection.
pH value: the instrument comprises the following steps: mettler
Detection by a Micro Pro pH meter.
Cation exchange chromatography CEX-HPLC:
residual amount of AS269 (small molecule toxin):
cell activity assay:
the cellular activity of the lyophilized samples relative to RS was examined using BT-474 cell line (ATCC, cat. HTB-20).
The specific process is as follows: BT-474 cells were cultured in a sufficient amount of ATCC hybrid-Care Medium (ATCC, cat. No. 46-X) containing 10% fetal bovine serum (Gibco, cat. No. 10099141) at 37 ℃ under 5% carbon dioxide, and seeded into 96-well cell culture plates (Corning, cat. No. 3599) at 1.5X 10 cells per well 4 BT-474 cells, 80. Mu.L/well. The lyophilized sample and RS were diluted from a gradient of 55.8. Mu.g/mL to 0.2. Mu.g/mL for 10 concentrations, 2 duplicate wells per dilution. The diluted lyophilized samples and RS were transferred to a plate inoculated with BT-474 cells at 10. Mu.L/well and incubated at 37 ℃ in 5% carbon dioxide. On day 5, 10. Mu.L of CCK-8 (Biyun, cat # C0043) detection reagent was added to each well, and the mixture was developed at 37 ℃ for 6 hours under 5% carbon dioxide, and the optical density values were read by an enzyme-linked microplate reader at wavelengths of 450nm and 620 nm. EC50 of the lyophilized samples and RS were calculated using SoftMax Pro software, and the relative cellular activities of the lyophilized samples were calculated using the following formula: relative cell activity (%) = (EC 50 RS)/(EC 50 lyophilized sample) × 100%.
Binding activity assay:
the binding activity of the lyophilized samples with respect to RS was tested by ELISA.
The specific process is as follows: HER2 antigen (R & D, cat # 1129-ER-050) was added to a 96-well plate (Corning, cat # 9018) at 50. Mu.L/well and coated overnight at 2-8 ℃; add blocking solution (Fisher Scientific, cat # 37528) at 200. Mu.L/well and shake at room temperature 200 + -10 rpm for 60min; diluting the lyophilized sample and RS from 10 μ g/mL gradient to 0.00017 μ g/mL with blocking solution in 96-well plate with total 11 concentrations of 2 multiple wells per dilution, shaking at room temperature 200 + -10 rpm for 60min; adding secondary antibody (Abcam, cat # ab 79115) at 50 μ L/well, and shaking at 200 + -10 rpm at room temperature for 60min; adding TMB color developing solution (Biyunyan, product number P0209) into 50 μ L/hole, shaking at room temperature of 200 + -10 rpm for 10min, adding 5% sulfuric acid into 50 μ L/hole, reading light absorption values at 450nm and 650nm respectively, calculating EC50 of lyophilized sample and RS by SoftMax Pro GxP software, and calculating relative binding activity of lyophilized sample by the following formula: relative binding activity (%) = (EC 50 RS)/(EC 50 lyophilized sample) × 100%.
Unless otherwise specified, the percentages used in the examples are percentages by mass.
EXAMPLE 1 stability Studies at different pH values
Formulations at different pH values were designed 1-10 (see Table 1).
TABLE 1 formulation of different pH values
Sample preparation:
(1) Respectively preparing 5mM acetate and histidine buffer solution, and adjusting to a target pH value by adopting hydrochloric acid or sodium hydroxide;
(2) Respectively preparing concentrated auxiliary material solutions by using a buffer salt solution, wherein the concentrations of polysorbate 80 and trehalose are respectively 14mg/ml (70 times of the target concentration) and 500mg/ml (10 times of the target concentration);
(3) Adding 10 parts of ADC solution into a pretreated ultrafiltration centrifugal tube (model: 30k, 15ml), centrifuging for 20min, respectively adding buffer salt solution, centrifuging for 20min, changing the solution, and repeating the solution changing and concentrating operation for four times to respectively obtain ADC concentrated solution containing target buffer salt;
(4) Measuring the protein concentration of the ADC solution after the solution change and concentration, adding a corresponding amount of the auxiliary material concentrated solution according to the calculated value, and fixing the volume to a target volume by adopting buffer salt;
(5) Filtering the sample, subpackaging, plugging and capping.
And (3) stability investigation:
the samples were left to stand at 25 ℃ for a total of 2 months. The results of the examination are shown in Table 2, in which T0 represents the time point when the sample started to be left, 25 ℃ to 1M represents the time point when the sample was left for one month, and 25 ℃ to 2M represents the time point when the sample was left for two months.
TABLE 2 summary of formulation stability examination results at different pH values
ND: since the AS269 remained growing more at this time point, CEX was not detected.
And (4) analyzing results:
AS269 residual results (table 2) show that, in each of the formulations (pH < 6.0) except formulation 3 (pH 6.0), formulation 4 (pH 6.3) and formulation 10 (pH 6.0), when examined at 25 ℃, a tendency of residual increase in AS269 was observed, and the lower the pH, the higher the residual AS269, indicating that the small molecule was easily dropped at a pH less than 6.0.
From the CEX results, it is clear that the decrease of the main CEX peak, the increase of the acidic peak and the basic peak are more likely to occur at pH > 6.0.
It follows that the pH of the lyophilized composition is critical to product stability, and that the pH of the lyophilized composition of the present invention is preferably between 5.7 and 6.3, more preferably around 6.0.
Example 2 formulation stability Studies at different ADC concentrations and polysorbate 80 concentrations
Based on the experimental results of example 1, formulation formulations 11-22 (table 3) were designed to examine the effect of different ADC concentrations and different polysorbate 80 concentrations on product stability. The samples are respectively sampled and detected after being placed at the temperature of 25 ℃ and shaken at 200rpm for 1 day and 3 days, and the detection results (table 4) show that the indexes of the samples in the formulas 11 to 22 before and after shaking are not obviously changed, so that the protein concentration is 5 to 30mg/ml and the polysorbate 80 concentration is 0.01 to 0.05 percent under the condition that the pH value is about 6.0, and the protein stability is good.
TABLE 3 formulation at different ADC concentrations and polysorbate 80 concentrations
Table 4 summary of the shake stability results for formulation formulations at different ADC concentrations and polysorbate 80 concentrations
Example 3 lyophilization parameter study of different solids content formulations
Formulation formulations 23-34 were designed to examine the effect of different combinations of anti-HER 2-ADC concentrations and trehalose concentrations on the lyophilization parameters (Tg'/Tc). The results in Table 5 show that there are differences in the lyophilization parameters for the different anti-HER 2-ADC and trehalose concentration combination formulations.
Considering the effect of lyophilization parameters (Tg'/Tc), lyophilization fill volume, and anti-HER 2-ADC concentration on lyophilization appearance, processing time, and stability, the anti-HER 2-ADC concentration can be 10-30mg/ml, and the trehalose concentration can be 2.5-8%. When lyophilized at anti-HER 2 ADC concentrations of 10mg/ml, 20mg/ml, and 30mg/ml, 2 (formula 23, formula 24), 3 (formula 27, formula 28, formula 29), and 1 (formula 31), respectively, the lyophilized product of formula 28 showed the best appearance (Table 6).
TABLE 5 summary of freeze-drying parameter results for different solid content formulations
Table 6 summary of freeze-dried appearance of different formulations
Example 4 Freeze drying Process study
The lyophilization process was examined using formulation 28 of example 3, namely 20mg/mL of anti-HER 2-ADC (ARX 788), 5mM histidine, 6wt.% trehalose, and 0.02wt.% polysorbate 80.
ADC samples and placebo (5 mM histidine, 6wt.% trehalose, and 0.02wt.% polysorbate 80) were formulated with reference to sample formulation steps (1) - (4) in example 1. The results are shown in Table 7. As can be seen from the results in table 7, the process parameters such as temperature, vacuum degree and the like of the first sublimation and the second sublimation affect the appearance, water content and other properties of the obtained freeze-dried product. The freeze-dried product obtained by the process 1' has unqualified appearance and high water content, and is not beneficial to the stability of the preparation; the appearance of the freeze-dried product obtained by the process 1 is greatly improved, but a certain proportion of unqualified products still exist; the processes 2-4 comprehensively consider and optimize the processes of primary sublimation and secondary sublimation, and the obtained freeze-dried product has excellent appearance and low water content, so that the yield and the product stability of the freeze-dried preparation can be remarkably improved.
TABLE 7 study of the lyophilization Process
Example 5 pilot production and stability study
Three batches of pilot production were carried out using the procedure-4 identified in example 4 (i.e., pilot batch-1 to pilot batch-3, batch size about 1500), the procedure was as follows:
(1) Taking the ADC stock solution out of the refrigerator 24 hours in advance, and standing at room temperature for thawing; after thawing, merging the stock solution into a Schottky bottle, and stirring and uniformly mixing;
(2) Sterilizing and filtering the uniformly mixed stock solution by two-stage sterile filters with the diameter of 0.22 mu m to obtain a sterile stock solution bag;
(3) Sterilizing the penicillin bottle by a tunnel oven and transferring the penicillin bottle into an isolator; the rubber plug and the aluminum cover are conveyed into the isolator through the RTP barrel after being subjected to steam sterilization;
(4) The sterile liquid storage bag is connected with the isolator through sterile connection and then filled into a sterilized penicillin bottle;
(5) After the penicillin bottles are half-stoppered, the penicillin bottles are conveyed into a freeze dryer, the plate layers of the freeze dryer are fully distributed, and empty penicillin bottles are used for supplementing the vacancy. Starting a program to freeze-dry, and performing full-pressure plugging after the freeze-drying is finished;
(6) Conveying the penicillin bottles subjected to the full-pressure plugging to a capping area for capping after being taken out of a box of a freeze dryer;
(7) And (5) cleaning the outer wall of the penicillin bottle after the cover is rolled, visually inspecting, labeling and packaging.
Production data show that three batches of production are completed smoothly, the appearance of the freeze-dried product is white or white-like blocks, the yield of the finished product is high, no defect products such as meltback or collapse exist, the redissolution time is within 1 minute, and the moisture content is below 1 percent.
With respect to the sterile powder for injection produced by the method of this example, the stability of three batches of samples for 24 months under long-term (5 ± 3 ℃) and 6 months under accelerated (25 ± 2 ℃) conditions was evaluated by using the appearance, pH, reconstitution time, moisture content, protein content, coupling ratio, SEC, CEX, CE-SDS, AS269 residue, cell activity, binding activity, and the like AS indices, and the results showed that there was no significant change in each index before and after the examination.
Meanwhile, one batch of samples (pilot plant batch-2) was subjected to high temperature influence factor investigation. The results show that the stability of the sample is very good, and the CEX peak (FIG. 2A) and the activity (FIG. 2B) change very little even if the sample is examined for 6 months under the condition of 40 ℃.
Unless otherwise defined, all terms used herein have the meanings commonly understood by those skilled in the art.
The described embodiments of the present invention are for illustrative purposes only and are not intended to limit the scope of the present invention, and those skilled in the art may make various other substitutions, alterations, and modifications within the scope of the present invention, and thus, the present invention is not limited to the above-described embodiments but only by the claims.
Claims (3)
1. A lyophilized formulation comprising an anti-HER 2-drug conjugate, wherein said lyophilized formulation is prepared by:
s1: cooling a freeze-dried composition containing an anti-HER 2-drug conjugate to 2-5 ℃ within 0.5-1.5 h, preserving heat for 0.5-1.5 h, then cooling to-50-40 ℃ within 3-5 h, preserving heat for 3-6 h, wherein the freeze-dried composition comprises 20mg/mL of ARX788, 5mM of histidine, 6wt.% of trehalose and 0.02wt.% of polysorbate 80, and adjusting the pH value to 6.0;
s2: raising the temperature of the freeze-dried composition treated in the step S1 to-23 to-20 ℃ within 0.2 to 0.8h, and keeping the temperature and the vacuum degree of 12 to 14Pa for 35 to 45h; and
s3: and (3) heating the freeze-dried composition treated in the step (S2) to 25-30 ℃ within 2-8 h, and keeping the temperature and the vacuum degree of 12-14 Pa for 5-10 h.
2. Use of the lyophilized formulation of claim 1 comprising an anti-HER 2-drug conjugate in the manufacture of a medicament for the treatment of cancer.
3. The use according to claim 2, wherein the cancer is breast cancer, gastric cancer, lung cancer, ovarian cancer, colorectal cancer, urothelial cancer, biliary tract cancer, liver cancer, brain cancer, esophageal cancer, endometrial cancer, uterine cancer, kidney cancer, bladder cancer, thyroid cancer, salivary gland cancer, or pancreatic cancer.
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| CN115708810A (en) * | 2022-11-03 | 2023-02-24 | 浙江新码生物医药有限公司 | Freeze-dried composition containing anti-CD 70-drug conjugate, freeze-dried preparation, preparation method and application thereof |
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| CN106456784A (en) * | 2014-04-16 | 2017-02-22 | 拜康有限公司 | Stable protein formulations comprising a molar excess of sorbitol |
| CN106729743B (en) * | 2015-11-23 | 2021-09-21 | 四川科伦博泰生物医药股份有限公司 | anti-ErbB 2 antibody-drug conjugate, and composition, preparation method and application thereof |
| CN110732023B (en) * | 2018-07-18 | 2023-06-16 | 江苏恒瑞医药股份有限公司 | HER2 antibody pharmaceutical composition and application thereof |
| WO2020233534A1 (en) * | 2019-05-17 | 2020-11-26 | 百奥泰生物制药股份有限公司 | Antibody-drug conjugate preparation, preparation method therefor and use thereof |
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