CN102988984A - Aqueous drug preparation of anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening stability - Google Patents
Aqueous drug preparation of anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening stability Download PDFInfo
- Publication number
- CN102988984A CN102988984A CN201210563488XA CN201210563488A CN102988984A CN 102988984 A CN102988984 A CN 102988984A CN 201210563488X A CN201210563488X A CN 201210563488XA CN 201210563488 A CN201210563488 A CN 201210563488A CN 102988984 A CN102988984 A CN 102988984A
- Authority
- CN
- China
- Prior art keywords
- preparation
- preferred
- concentration
- pharmaceutical preparation
- sodium
- 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.)
- Granted
Links
Abstract
The invention provides a new liquid aqueous drug preparation of an anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening the stability, wherein antibody solution is stabilized and osmotic pressure is regulated by sorbitol; disodium hydrogen phosphate/citric acid monohydrate or sodium acetate/acetic acid is taken as a buffer agent, and preferably, the disodium hydrogen phosphate/citric acid monohydrate is taken as the buffer agent; and tween-20 is taken as a surfactant, so that antibody preparation is stable, and long in retention time, meanwhile, preparation components are simplified, and the preparation is prepared simply and conveniently.
Description
Technical field
The invention belongs to biological technical field, relate to the liquid aqueous pharmaceutical preparation of total man's monoclonal antibody drug of enhanced stability; Relate to the purposes of using liquid aqueous pharmaceutical preparation treatment relevant disease of the present invention.
Background technology
Huamn tumor necrosis factory alpha (TNF-α) has been proved relevant with various diseases, such as autoimmune diseasees such as rheumatoid arthritis, psoriasises.A plurality of TNF alpha antibody medicines are used for the treatment of the relevant autoimmune class disease of TNF-α by drugs approved by FDA, such as adalimumab (Adalimumab, trade name Humira, Abbott).
The adalimumab that is applied to clinical treatment is the liquid infusion preparation, and interior packaging material is precharging type syringe.Its preparation prescription comprises sodium chloride, sodium hydrogen phosphate, sodium dihydrogen phosphate, citric acid, sodium citrate, mannitol, anhydrous sorbitol polyoxyethylene ether monoleate (Tween 80), water for injection (Humira description).Although the said preparation existence and stability, but still be not enough to long-time stabilization of antibodies, the expectation finished product should not be placed for more time.This may be because the employed antioxidant of said preparation and osmotic pressure regulator---mannitol can not be brought into play good Stabilization.In addition, the surfactant that existing adalimumab uses is tween 80, although thereby it can suppress and/or stop bad result's stabilization of antibodies such as the polymerization of protein molecular on water and Air Interface, interior packaging material contact surface, absorption, thereby produce acid, aldehyde, hydrogen peroxide but unsaturated bond oxidation and ester linkage hydrolyzing also can occur to a certain extent himself, thereby affected the stability of antibody.Simultaneously, its prescription buffer solution composition is redundant complicated, has not only increased product cost, and has made the formulation operations of preparation loaded down with trivial details.
Summary of the invention
In order to address the above problem, the invention provides a kind of liquid aqueous pharmaceutical preparation of new TNF alpha antibody human monoclonal antibodies, make antibody preparation more stable, the holding time is longer; Simplified simultaneously the preparation composition, made the preparation of preparation more simple and convenient.
Aqueous medicament preparations provided by the invention comprises:
(a) TNF alpha antibody human monoclonal antibodies;
(b) polyhydric alcohol;
(c) buffer agent;
(d) surfactant;
(e) sodium chloride;
(f) water;
Wherein, described polyhydric alcohol is sorbitol; Described buffer agent is sodium hydrogen phosphate/monohydrate potassium or sodium acetate/acetic acid, preferably phosphoric acid disodium hydrogen/monohydrate potassium; Described surfactant is polyoxyethylene sorbitan monolaurate (tween 20).
The pH of liquid aqueous pharmaceutical preparation of the present invention is 4-8, and electric conductance is 8-18mS/cm, and osmotic pressure is 250-400mOsm/kg.
Liquid aqueous pharmaceutical preparation of the present invention, it comprises 10-80g/L restructuring TNF alpha antibody human monoclonal antibodies, 10-20g/L sorbitol, 0.01-3g/L tween 20,2-14g/L sodium chloride, and the buffer agent of sodium hydrogen phosphate/monohydrate potassium, its pH is 4-8, preferred pH 5-6, more preferably pH 5.0-5.4.
Liquid aqueous pharmaceutical preparation of the present invention, it comprises 10-80g/L restructuring TNF alpha antibody human monoclonal antibodies, 10-20g/L sorbitol, 0.01-3g/L tween 20,2-14g/L sodium chloride, and the buffer agent of sodium acetate/acetic acid, and its pH is 4-8, preferred pH 5-6, more preferably pH 5.0-5.4.
The specific embodiment
For the ease of understanding the present invention, some term of the present invention is defined:
Term " pharmaceutical preparation " represents its form so that the biologic activity of active component is clearly effective, and it does not comprise obvious other poisonous composition to the experimenter who uses described preparation.
" stable " preparation is preparation when preserving, and antibody wherein can keep its physical stability and/or chemical stability and/or biological stability basically.
" buffer solution ": when adding a certain amount of bronsted lowry acids and bases bronsted lowry in some solution, have and hinder the effect that pH value of solution changes, be called cushioning effect, such solution is called buffer solution.
" buffer agent ": can make pH keep within the specific limits substantially invariable material.
Within the scope of the invention, " the treatment effective dose " of antibody or " effective dose " are illustrated in the effective dose of prevention or treatment disease aspect, and for the treatment of described disease, described antibody is effective.
Term " humanTNF-α " expression human cell factor, it is to exist with the secreted form of 17kD and 26kD film association form, the biologic activity form comprises the trimer of covalently bound 17kD molecule.Its concrete structure sees Pennica, and D. waits (1984) Nature 312:724-729; Davis, J.M. waits (1987) Biochemistry26:1322-1326; And Jones, E.Y. waits (1989) Nature 338:225-228.
Term " recombinant human antibody " is intended to comprise recombination method preparation, people's antibody of expressing, producing or separating.
Stock solution: the aqueous medicament preparations that does not contain tween.
Finished product: the aqueous medicament preparations that adds according to quantity tween.
SEC-HPLC: size-exclusion high performance liquid chromatography.
CEX-HPLC (Acidic, Main, Basic): cation exchange-high performance liquid chromatography (acid peak, main peak, alkaline peak).
NR-CE-SDS: non-reduced high performance capillary electrophoresis.
Potency: biologic activity.
The invention provides aqueous medicament preparations, comprising:
(a) TNF alpha antibody human monoclonal antibodies;
(b) polyhydric alcohol;
(c) buffer agent;
(d) surfactant;
(e) sodium chloride;
(f) water;
Wherein, described polyhydric alcohol is sorbitol; Described buffer agent is sodium hydrogen phosphate/monohydrate potassium or sodium acetate/acetic acid, preferably phosphoric acid disodium hydrogen/monohydrate potassium; Described surfactant is polyoxyethylene sorbitan monolaurate (tween 20).
The pH of liquid aqueous pharmaceutical preparation of the present invention is 4-8, and electric conductance is 8-18mS/cm, and osmotic pressure is 250-400mOsm/kg.
Aqueous medicament preparations of the present invention contains the restructuring TNF alpha antibody human monoclonal antibodies for the treatment of effective dose, and its concentration is 10-80g/L, preferred 20-70g/L, particularly preferably 50g/L.
Known polyhydric alcohol can play antioxidation, therefore can stabilization of antibodies.The present invention uses sorbitol, and concentration is 10-20g/L, preferred 12-18g/L, and 15g/L most preferably in the embodiments of the invention, adopts and uses the mannitol can better stabilization of antibodies in the existing adalimumab preparation of sorbitol.
Aqueous medicament preparations pH of the present invention is 4-8, preferred 5-6, particularly preferably 5.2.Buffer agent sodium hydrogen phosphate/monohydrate potassium of the present invention or sodium acetate/acetic acid, be preferably sodium hydrogen phosphate/monohydrate potassium, the molar concentration scope of the buffer solution of its pH 5.2 is 10-40mM, preferred 21.5mM, be sodium hydrogen phosphate 2g/L, monohydrate potassium 1.54g/L, and regulate final preparation pH 5.2 with sodium hydroxide.Use this buffer agent can make with original adalimumab preparation in the buffer solution of each ion same concentrations, guarantee that phosphate radical, citrate, sodium ion change all in 5%, simplified the preparation process for preparation, saved cost.Buffer agent in one embodiment of the present of invention is sodium acetate/acetic acid, and the molar concentration scope of its pH 5.2 buffer solution is 10-40mM, preferred 18.5mM, i.e. and sodium acetate 1.23g/L, acetic acid 0.2mL/L, and regulate final preparation pH 5.2 with acetic acid.
Inhibitory action is played in the polymerization of polyoxyethylene sorbitan fatty acid ester (tween) antagonist, and the present invention uses the less tween 20 of unsaturated bond (polyoxyethylene sorbitan monolaurate) thereby reduces the adverse effect of antagonist stability to reduce autoxidation.In an embodiment, adopt the more original adalimumab preparation of tween 20 to adopt tween 80 (polyoxyethylene sorbitan monooleate dehydration) can make pharmaceutical preparation that better stability is arranged.Especially, in a stability (illumination experiment) embodiment, when using the buffer agent of sodium hydrogen phosphate/monohydrate potassium, tween 20 can significantly improve the stability of pharmaceutical preparation.Optional tween 20 concentration is 0.01-3g/L, preferred 1g/L.
Sodium chloride is osmotic pressure regulator, and in embodiment of the present invention, its concentration is 2-14g/L, preferred 3-12g/L, most preferably 6.16g/L.
The buffer solution of liquid aqueous pharmaceutical preparation of the present invention is sodium hydrogen phosphate/monohydrate potassium solution, pH 4-8, preferred pH 5-6, more preferably pH 5.0-5.4, electric conductance is 8-18mS/cm, preferred 11.1-14.1mS/cm, osmotic pressure is 250-400mOsm/kg, preferred 280-360mOsm/kg.
The buffer solution of liquid aqueous pharmaceutical preparation of the present invention is sodium acetate/acetic acid solution, and pH is 4-8, preferred pH 5-6, more preferably pH 5.0-5.4, electric conductance is 8-18mS/cm, preferred 9.6-13.0mS/cm, osmotic pressure is 250-400mOsm/kg, preferred 270-350mOsm/kg.
The invention provides a kind of liquid aqueous pharmaceutical preparation of new TNF alpha antibody human monoclonal antibodies, make antibody preparation more stable, the holding time is longer; Simplified simultaneously the preparation composition, made the preparation of preparation more simple and convenient.
Describe the present invention in detail referring to embodiment, should be understood that following embodiment is intended to explanation, is not construed as limiting the present invention.
The TNF alpha antibody human monoclonal antibodies that the present invention uses is the IgG1 recombinant antibodies D2E7 (referring to Chinese patent ZL97193635.8) of Genor Biopharma Co., Ltd.'s preparation; Water is the self-produced water for injection of Genor Biopharma Co., Ltd.; Sorbitol is available from Shijiazhuang City, Hebei Province timely snow pharmaceutical Co. Ltd, and it meets the Chinese Pharmacopoeia pharmaceutical grade; Other reagent is all available from J.T.Baker company, and it meets the American Pharmacopeia pharmaceutical grade.
It below is instrument model used in the present invention: balance: Sartorius TE4101; PH and electrical conductivity integrated instrument: METTLER TOLEDO SevenMulti; Osmotic tester: Model3250Osmometer (ADVANCED INSTRUMENTS, INC.); HPLC:Waters2695-2487, wherein, the used analytical column of SEC-HPLC is Tosoh TSKgel G3000SW; The used analytical column of CEX-HPLC is Tosoh TSKgel CM-STAT; It is Beckman PA800plus that NR-CE-SDS measures instrument.
The preparation of embodiment 1. buffer solution
1.1 preparation 8L sodium hydrogen phosphate/monohydrate potassium ultrafiltration buffer solution (ultrafiltration buffer solution A)
Take by weighing respectively 16g sodium hydrogen phosphate, 12.32g monohydrate potassium, 120g sorbitol, 49.28g sodium chloride, add to 7.9L with water for injection, until dissolving fully, regulating pH 5.2 with the 1M sodium hydroxide behind the mix homogeneously, it is for subsequent use to be settled to 8.0L.
1.2 preparation 8L sodium hydrogen phosphate/sodium dihydrogen phosphate and sodium citrate/citric acid ultrafiltration buffer solution (ultrafiltration buffer solution B)
Take by weighing respectively 6.88g two hypophosphite monohydrate sodium dihydrogens, 12.24g two hypophosphite monohydrate disodium hydrogens, 2.4g sodium citrate, 10.4g monohydrate potassium, 96g mannitol, 49.28g sodium chloride, add to 7.9L with water for injection, until dissolving fully, regulating pH 5.2 with the 1M sodium hydroxide behind the mix homogeneously, it is for subsequent use to be settled to 8.0L.
1.3 preparation 8L sodium acetate/acetic acid ultrafiltration buffer solution (ultrafiltration buffer solution C)
Take by weighing respectively 9.84g sodium acetate, 120g sorbitol, 49.28g sodium chloride, water for injection adds to 7.9L, until dissolve fully, behind the mix homogeneously with second acid for adjusting pH 5.2, it is for subsequent use to be settled to 8.0L.
Embodiment 2. antibody concentration are the preparation of the ultrafiltration and concentration liquid of 51g/L
2.1 buffer solution is the preparation (ultrafiltration and concentration liquid A) of the ultrafiltration and concentration liquid of sodium hydrogen phosphate/monohydrate potassium solution
Ultrafiltration instrument VIVA Flow 200 (Sartorius of producer) cleans with the 0.2M sodium hydroxide by the manufacturer's recommended step, comprise and clean ultrafilter membrane, ultrafiltration cup, pipeline, outlet etc., clean to remove sodium hydroxide with the ultrafiltration buffer solution A among the 0.5L embodiment 1.1 again.0.48L TNF alpha antibody human monoclonal antibodies solution (antibody concentration 25g/L, total antibody 12g) is added in the ultrafiltration cup, begin equal-volume with ultrafiltration buffer solution A and change liquid, change the long-pending 4.8L of being of liquid.Change liquid complete after, antibody-solutions is concentrated into about 0.2L from 0.48L, reclaim antibody-solutions after the depolarization, the antibody-solutions volume that reclaims after measured is 0.204L, concentration is 52g/L (meet the about 0.2L of volume, concentration requires greater than 51g/L).The antibody-solutions that will add after 4mL ultrafiltration buffer solution A will reclaim is diluted to 51g/L, with aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, the filtering solution Preservation in sterile condition that obtains.
2.2 buffer solution is the preparation (ultrafiltration and concentration liquid B) of the ultrafiltration and concentration liquid of sodium hydrogen phosphate/sodium dihydrogen phosphate and sodium citrate/citric acid solution
Ultrafiltration instrument VIVA Flow 200 basic operations clean to remove sodium hydroxide with the ultrafiltration buffer solution B among the 0.5L embodiment 1.2 again with described in the embodiment 2.1.0.48L TNF alpha antibody human monoclonal antibodies solution (antibody concentration 25g/L, total antibody 12g) is added in the ultrafiltration cup, begin equal-volume with ultrafiltration buffer solution B and change liquid, change the long-pending 4.8L of being of liquid.Change liquid complete after, antibody-solutions is concentrated into about 0.2L from 0.48L, reclaim antibody-solutions after the depolarization, the antibody-solutions volume that reclaims after measured is 0.209L, concentration is 52g/L (meet the about 0.2L of volume, concentration requires greater than 51g/L).The antibody-solutions that will add after 4mL ultrafiltration buffer solution B will reclaim is diluted to 51g/L, with aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, the filtering solution Preservation in sterile condition that obtains.
2.3 buffer solution is the preparation (ultrafiltration and concentration liquid C) of the ultrafiltration and concentration liquid of sodium acetate/acetic acid solution
Ultrafiltration instrument VIVA Flow 200 basic operations clean to remove sodium hydroxide with the ultrafiltration buffer solution C among the 0.5L embodiment 1.3 again with described in the embodiment 2.1.0.48L TNF alpha antibody human monoclonal antibodies solution (antibody concentration 25g/L, total antibody 12g) is added in the ultrafiltration cup, begin equal-volume with ultrafiltration buffer solution C and change liquid, change the long-pending 4.8L of being of liquid.Change liquid complete after, antibody-solutions is concentrated into about 0.2L from 0.48L, reclaim antibody-solutions after the depolarization, the antibody-solutions volume that reclaims after measured is 0.195L, concentration is 54g/L (meet the about 0.2L of volume, concentration requires greater than 51g/L).The antibody-solutions that will add after 11mL ultrafiltration buffer solution C will reclaim is diluted to 51g/L, with aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, the filtering solution Preservation in sterile condition that obtains.
The preparation of embodiment 3. stock solutions
3.1 preparation 0.9mL/ cillin bottle stock solution A (buffer solution is sodium hydrogen phosphate/monohydrate potassium, does not contain the preparation of tween)
Under the aseptic condition, get the ultrafiltration and concentration liquid A of 50mL embodiment 2.1, add the ultrafiltration buffer solution A with the 1mL embodiment 1.1 of aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, fully mix homogeneously is stock solution A.Under the aseptic condition, it is sub-packed in the aseptic apyrogenic liquid infusion agent cillin bottle of 2mL, loading amount 0.9mL/ bottle, 54 of packing, 2-8 ℃ keeps in Dark Place stand-by behind the Zha Gai that jumps a queue.
Every situation explanation: every loading amount 0.9mL, labelled amount 0.8mL, it is composed as follows:
3.2 preparation 0.9mL/ cillin bottle stock solution B (buffer solution is sodium hydrogen phosphate/sodium dihydrogen phosphate and sodium citrate/citric acid solution, does not contain the preparation of tween 80)
Under the aseptic condition, get the ultrafiltration and concentration liquid B of 50mL embodiment 2.2, add the ultrafiltration buffer solution B with the 1mL embodiment 1.2 of aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, fully mix homogeneously is stock solution B.Under the aseptic condition, it is sub-packed in the aseptic apyrogenic liquid infusion agent cillin bottle of 2mL, loading amount 0.9mL/ bottle, 54 of packing, 2-8 ℃ keeps in Dark Place stand-by behind the Zha Gai that jumps a queue.
Every situation explanation: every loading amount 0.9mL, labelled amount 0.8mL, it is composed as follows:
3.3 preparation 0.9mL/ cillin bottle stock solution C (buffer solution is sodium acetate/acetic acid solution, does not contain the preparation of tween)
Under the aseptic condition, get the ultrafiltration and concentration liquid C of 50mL embodiment 2.3, add the ultrafiltration buffer solution C with the 1mL embodiment 1.3 of aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, fully mix homogeneously is stock solution C.Under the aseptic condition, it is sub-packed in the aseptic apyrogenic liquid infusion agent cillin bottle of 2mL, loading amount 0.9mL/ bottle, 54 of packing, 2-8 ℃ keeps in Dark Place stand-by behind the Zha Gai that jumps a queue.
Every situation explanation: every loading amount 0.9mL, labelled amount 0.8mL, it is composed as follows:
The preparation of embodiment 4. finished products
4.1 preparation 0.9mL/ cillin bottle finished product A (buffer solution is sodium hydrogen phosphate/monohydrate potassium, contains the preparation of tween 20)
Take by weighing the 1.00g tween 20, in 19.6mL, with aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, the 10-19mL Preservation in sterile condition of getting after the filtration is for subsequent use, is 51 * polysorbas20 mother solution with the water for injection standardize solution.
Under the aseptic condition, get the ultrafiltration and concentration liquid A of 50mL embodiment 2.1, add 1mL 51 * polysorbas20 mother solution, fully mix homogeneously is finished product A.Under the aseptic condition, it is sub-packed in the aseptic apyrogenic liquid infusion agent cillin bottle of 2mL, loading amount 0.9mL/ bottle, 54 of packing, 2-8 ℃ keeps in Dark Place stand-by behind the Zha Gai that jumps a queue.
Every situation explanation: every loading amount 0.9mL, labelled amount 0.8mL, it is composed as follows:
4.2 preparation 0.9mL/ cillin bottle finished product B (buffer solution is sodium hydrogen phosphate/sodium dihydrogen phosphate and sodium citrate/citric acid solution, contains the preparation of tween 80)
Take by weighing the 1.00g Tween 80, in 19.6mL, with aseptic apyrogenic 0.22 μ m aseptic filtration membrane filtration, the 10-19mL Preservation in sterile condition of getting after the filtration is for subsequent use, is 51 * tween 80 mother solution with the water for injection standardize solution.
Under the aseptic condition, get the ultrafiltration and concentration liquid B of 50mL embodiment 2.2, add 1mL 51 * tween 80 mother solution, fully mix homogeneously is finished product B.Under the aseptic condition, it is sub-packed in the aseptic apyrogenic liquid infusion agent cillin bottle of 2mL, loading amount 0.9mL/ bottle, 54 of packing, 2-8 ℃ keeps in Dark Place stand-by behind the Zha Gai that jumps a queue.
Every situation explanation: every loading amount 0.9mL, labelled amount 0.8mL, it is composed as follows:
4.3 preparation 0.9mL/ cillin bottle finished product C (buffer solution is sodium acetate/acetic acid solution, contains the preparation of tween 20)
Under the aseptic condition, get the ultrafiltration and concentration liquid C of 50mL embodiment 2.3, add the 51 * tween 20 mother solution that makes among the 1mL embodiment 4.1, fully mix homogeneously is finished product C.Under the aseptic condition, it is sub-packed in the aseptic apyrogenic liquid infusion agent cillin bottle of 2mL, loading amount 0.9mL/ bottle, 54 of packing, 2-8 ℃ keeps in Dark Place stand-by behind the Zha Gai that jumps a queue.
Every situation explanation: every loading amount 0.9mL, labelled amount 0.8mL, it is composed as follows:
Embodiment 5. study on the stability
5.15 ± 3 ℃ of long-time stability experiments and 25 ± 2 ℃ of acceleration experiments
Get respectively each 12 of 0.9mL/ cillin bottle stock solution A, B, C and finished product A, B in above-described embodiment, C, be placed in 5 ± 3 ℃ the Scientific Revco cold storage refrigerator, expectation is respectively at the 0th day, the 5th day, the 15th day, the 30th day, the 3rd month, the 6th month sample analysis, and every kind of each sample point of sample is put 2; Get respectively each 12 of 0.9mL/ cillin bottle stock solution A, B, C and finished product A, B in above-described embodiment, C, be placed in 25 ± 2 ℃ the Climacell 222 constant temperature and humidity incubators, respectively at the 0th day, the 5th day, the 15th day, the 30th day, the 3rd month, the 6th month sample analysis, every kind of each sample point of sample is put 2.Analytical method is carried out the detection of SEC-HPLC, CEX-HPLC, NR-CE-SDS and biologic activity again for immediately naked eyes detection outward appearance after taking a sample.In the above-mentioned experimental design, if the 30th day, the 6th month result meets expection, then needn't analyze the 15th day, the 3rd month sample point.Experimental result sees Table 1 to table 7.
The 0th day testing result of table 1.
Table 2.5 ± 3 ℃, the 5th day long-time stability experimental result
Table 3.5 ± 3 ℃, the 30th day long-time stability experimental result
Table 4.5 ± 3 ℃, the 6th month long-time stability experimental result
Table 5.25 ± 2 ℃ were accelerated experimental result on the 5th day
Table 6.25 ± 2 ℃ were accelerated experimental result on the 30th day
Table 7.25 ± 2 ℃ were accelerated experimental result on the 6th month
Experimental result shows, in 5 ± 3 ℃ of groups, stock solution A, B, C and finished product A, B, C the 5th day, the 30th day, the 6th month all testing result all with the 0th day no significant difference, use buffer agent sodium hydrogen phosphate/monohydrate potassium with tween 20 and use buffer agent sodium acetate/acetic acid and tween 20 can reach the stability identical with the agent of A Da wood monoclonal antibody liquid infusion.
In 25 ± 2 ℃ of groups, stock solution A, B, C and finished product A, B, C was at the 5th day, all testing results of the 30th day all with the 0th day no significant difference, but 6th month testing result shows difference (as shown in table 7): by relatively finding, the SEC-HPLC purity of finished product A is than finished product B, the variation at C high about 1% and main peak oxytropism peak is than finished product B, little about 7-8% of C, this explanation finished product A is better than finished product B and C, further, finished product A, B, the SEC-HPLC purity of C is higher than respectively its stock solution A, B, the purity of C about 1%, and the variation at corresponding generation main peak oxytropism peak still less, illustrate that tween has the effect of obvious stabilization of antibodies, particularly prevents the polymerization of antibody molecule; Find that by contrasting 3 stock solution groups used buffer agent sodium hydrogen phosphate/monohydrate potassium has better Stabilization than buffer agent sodium acetate/acetic acid antagonist in the present embodiment, used sorbitol has better Stabilization than mannitol antagonist.
5.2 illumination experiment
Get respectively each 8 of 0.9mL/ cillin bottle stock solution A, B, C and finished product A, B in above-described embodiment, C, wrap up lucifuge with aluminium foil, then be placed in 40 ± 2 ℃ the Climacell 404 constant temperature and humidity incubators, respectively at the 0th day, the 5th day, the 15th day, the 30th day every kind of sample taken a sample respectively two and analyze; Get respectively each 8 of 0.9mL/ cillin bottle stock solution A, B, C and finished product A, B in above-described embodiment, C, be placed in 40 ± 2 ℃, the Climacell 404 constant temperature and humidity incubators of illumination (intensity 4000-5000lx), respectively at the 0th day, the 5th day, the 15th day, the 30th day every kind of sample taken a sample respectively two and analyze.Analytical method is carried out the detection of SEC-HPLC, CEX-HPLC, NR-CE-SDS and biologic activity again for immediately naked eyes detection outward appearance after taking a sample.In the above-mentioned experimental design, if the 30th day result meets expection, then needn't analyze the 15th day sample point.Experimental result sees Table 8 to table 10.
Table 8.40 ± 2 ℃, 5 days influence factor's experimental results of lucifuge
Table 9.40 ± 2 ℃, 30 days influence factor's experimental results of lucifuge
Table 10.40 ± 2 ℃, 5 days influence factor's experimental results of illumination
Under the aCEX detection, peak shape produces and to move to right, and accurate integration is so data are unlisted.
1) in 40 ± 2 ℃ of lucifuge groups, stock solution A, B, C and finished product A, B, C are showed no the 5th day, the 30th day outward appearance unusually.Along with time lengthening; SEC-HPLC and NR-CE-SDS purity all descend to some extent; but stock solution A, C and finished product A, C purity fall are less than stock solution B and finished product B; illustrate that stock solution A, C and finished product A, C are better than stock solution B and finished product B, supposition may be that the sorbitol of higher concentration has better protective effect than mannitol antagonist molecule.Simultaneously, along with the prolongation of time, the main peak peak area of the antibody in each sample constantly reduces, and acid peak-to-peak area constantly increases, and the part main peak is transformed into acid peak.In the same time, it is more that main peak is transformed into acid peak, it is more to illustrate that antibody itself changes, more unstable, further 6 samples of contrast can find among finished product A, C and stock solution A, the C by the increase at main peak oxytropism peak less than among finished product B and the stock solution B by the increase at main peak oxytropism peak, illustrate that finished product A, C and stock solution A, C are more conducive to stablizing of antibody than finished product B and stock solution B.In whole experimental period, biologic activity does not detect decline.
2) 40 ± 2 ℃ of illumination (intensity 4000-5000lx) group was at the 5th day outward appearance visible particle shape material, SEC-HPLC and NR-CE-SDS purity all decline to a great extent, the about 20-35% of amplitude, CEX-HPLC can't detect, and biologic activity sharply descends and loses analysis significance.The 10th day visible obvious liquid-solid segregation phenomenon of outward appearance, therefore investigated the meaning that sample has not had analysis in the 15th day, 30 days.
Conclusion:
The SEC-HPLC of each stock solution, finished product and NR-CE-SDS purity show that order is as shown in table 11 from excellent to bad:
Table 11.40 ± 2 ℃ illumination effect Factor Experiment result
Annotate:＞＞＞expression significantly better than, the former purity is high by 5% than the latter;＞＞expression is better than, and the former purity is than the high 2-5% of the latter;＞expression slightly is better than, and the former purity is than the high 1-2% of the latter;=expression is equal to, both purity differ ± 1% in.
Liquid infusion preparation provided by the invention---buffer agent is sodium hydrogen phosphate-citric acid or sodium acetate/acetic acid as can be drawn from Table 11, and surfactant is tween 20---aspect stable, be better than existing adalimumab liquid infusion preparation, preferred buffer system is that sodium hydrogen phosphate-citric acid and surfactant are the liquid infusion preparation of tween 20, and its stability significantly is better than existing adalimumab liquid infusion preparation.
Claims (10)
1. liquid aqueous pharmaceutical preparation comprises:
(a) TNF alpha antibody human monoclonal antibodies;
(b) polyhydric alcohol;
(c) buffer agent;
(d) surfactant;
(e) sodium chloride;
(f) water;
Wherein, described polyhydric alcohol is sorbitol; Described buffer agent is sodium hydrogen phosphate/monohydrate potassium or sodium acetate/acetic acid; Described surfactant is polyoxyethylene sorbitan monolaurate.
2. the liquid aqueous pharmaceutical preparation of claim 1, wherein said TNF alpha antibody human monoclonal antibodies is the D2E7 monoclonal antibody, its concentration is 10-80g/L, preferred 20-70g/L, particularly preferably 50g/L.
3. the liquid aqueous pharmaceutical preparation of claim 1, the concentration of wherein said sorbitol is 10-20g/L, preferred 12-18g/L, most preferably 15g/L.
4. the liquid aqueous pharmaceutical preparation of claim 1, the concentration of wherein said polyoxyethylene sorbitan monolaurate is 0.01-3g/L, preferred 1g/L.
5. the liquid aqueous pharmaceutical preparation of claim 1, wherein said buffer agent is sodium hydrogen phosphate/monohydrate potassium, pH is 5-6, preferred pH 5.0-5.4, more preferably pH 5.2, and the molar concentration scope of the buffer solution of its pH5.2 is 10-40mM, preferred 21.5mM, be sodium hydrogen phosphate 2g/L, monohydrate potassium 1.54g/L, and regulate final preparation pH 5.2 with sodium hydroxide.
6. the liquid aqueous pharmaceutical preparation of claim 1, wherein said buffer agent is sodium acetate/acetic acid, pH is 5-6, preferred pH 5.0-5.4, more preferably pH 5.2, and the molar concentration scope of its pH 5.2 buffer solution is 10-40mM, preferred 18.5mM, be sodium acetate 1.23g/L, acetic acid 0.2mL/L, and regulate final preparation pH 5.2 with acetic acid.
7. each liquid aqueous pharmaceutical preparation of claim 1-6, the concentration of wherein said sodium chloride is 2-14g/L, preferred 3-12g/L, most preferably 6.16g/L.
8. the liquid aqueous pharmaceutical preparation of claim 7, wherein said TNF alpha antibody human monoclonal antibodies is the D2E7 monoclonal antibody, concentration is 10-80g/L, preferred 20-70g/L, particularly preferably 50g/L; The concentration of described sorbitol is 10-20g/L, preferred 12-18g/L, most preferably 15g/L; Described surfactant is polyoxyethylene sorbitan monolaurate, and concentration is 0.01-3g/L, preferred 1g/L; Described buffer agent is sodium hydrogen phosphate/monohydrate potassium or sodium acetate/acetic acid, and preferably phosphoric acid disodium hydrogen/monohydrate potassium is buffer agent, and the pH of described aqueous medicament preparations is 5-6, preferred pH 5.0-5.4, and more preferably pH 5.2.
9. the liquid aqueous pharmaceutical preparation of claim 8, it is the aqueous solution of following material:
10. the liquid aqueous pharmaceutical preparation of claim 8, it is the aqueous solution of following material:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210563488.XA CN102988984B (en) | 2012-12-21 | 2012-12-21 | Aqueous drug preparation of anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening stability |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210563488.XA CN102988984B (en) | 2012-12-21 | 2012-12-21 | Aqueous drug preparation of anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening stability |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102988984A true CN102988984A (en) | 2013-03-27 |
| CN102988984B CN102988984B (en) | 2015-05-20 |
Family
ID=47918418
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210563488.XA Active CN102988984B (en) | 2012-12-21 | 2012-12-21 | Aqueous drug preparation of anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening stability |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102988984B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2863951A1 (en) * | 2012-06-12 | 2015-04-29 | Boehringer Ingelheim International GmbH | Pharmaceutical formulation for a therapeutic antibody |
| US9682145B2 (en) | 2012-09-07 | 2017-06-20 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| CN107619442A (en) * | 2016-07-15 | 2018-01-23 | 上海百迈博制药有限公司 | One kind recombinates the anti-full human monoclonal antibody preparations of TNF α |
| CN109239335A (en) * | 2018-09-11 | 2019-01-18 | 广州万孚生物技术股份有限公司 | Joint inspection test strips and preparation method thereof |
| CN110151988A (en) * | 2018-02-11 | 2019-08-23 | 百奥泰生物制药股份有限公司 | A kind of human antibody preparation of targeted therapy TNF-α related disease |
| CN112656939A (en) * | 2014-09-22 | 2021-04-16 | 正大天晴药业集团股份有限公司 | Pharmaceutical composition of humanized antibody for vascular endothelial growth factor |
| US11071782B2 (en) | 2016-04-20 | 2021-07-27 | Coherus Biosciences, Inc. | Method of filling a container with no headspace |
| US11229702B1 (en) | 2015-10-28 | 2022-01-25 | Coherus Biosciences, Inc. | High concentration formulations of adalimumab |
| CN116239678A (en) * | 2023-05-12 | 2023-06-09 | 广州悦洋生物技术有限公司 | Antioxidant composition, application, protective agent and solution |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1256148A (en) * | 1999-12-06 | 2000-06-14 | 天津华立达生物工程有限公司 | Stable water solution of interferon |
| CN1688339A (en) * | 2002-08-16 | 2005-10-26 | 艾博特生物技术有限公司 | Formulation of human antibodies for treating TNF-alpha associated disorders |
| CN101896199A (en) * | 2007-11-30 | 2010-11-24 | 株式会社绿十字 | Pharmaceutical formulation comprising hepatitis B virus neutralizing human antibody |
| CN102327612A (en) * | 2010-07-13 | 2012-01-25 | 上海张江生物技术有限公司 | Steady humanized monoclonal antibody preparation |
| CN102458469A (en) * | 2009-05-04 | 2012-05-16 | 艾博特生物技术有限公司 | Stable high protein concentration formulations of human anti-tnf-alpha-antibodies |
-
2012
- 2012-12-21 CN CN201210563488.XA patent/CN102988984B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1256148A (en) * | 1999-12-06 | 2000-06-14 | 天津华立达生物工程有限公司 | Stable water solution of interferon |
| CN1688339A (en) * | 2002-08-16 | 2005-10-26 | 艾博特生物技术有限公司 | Formulation of human antibodies for treating TNF-alpha associated disorders |
| CN101896199A (en) * | 2007-11-30 | 2010-11-24 | 株式会社绿十字 | Pharmaceutical formulation comprising hepatitis B virus neutralizing human antibody |
| CN102458469A (en) * | 2009-05-04 | 2012-05-16 | 艾博特生物技术有限公司 | Stable high protein concentration formulations of human anti-tnf-alpha-antibodies |
| CN102327612A (en) * | 2010-07-13 | 2012-01-25 | 上海张江生物技术有限公司 | Steady humanized monoclonal antibody preparation |
Cited By (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2863951A1 (en) * | 2012-06-12 | 2015-04-29 | Boehringer Ingelheim International GmbH | Pharmaceutical formulation for a therapeutic antibody |
| JP2015519382A (en) * | 2012-06-12 | 2015-07-09 | ベーリンガー インゲルハイム インターナショナル ゲゼルシャフト ミット ベシュレンクテル ハフツング | Pharmaceutical formulations for therapeutic antibodies |
| US10716852B2 (en) | 2012-09-07 | 2020-07-21 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10772959B2 (en) | 2012-09-07 | 2020-09-15 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10716854B2 (en) | 2012-09-07 | 2020-07-21 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9724415B2 (en) | 2012-09-07 | 2017-08-08 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10722579B2 (en) | 2012-09-07 | 2020-07-28 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9731009B2 (en) | 2012-09-07 | 2017-08-15 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9737600B2 (en) | 2012-09-07 | 2017-08-22 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9757454B2 (en) | 2012-09-07 | 2017-09-12 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10799585B2 (en) | 2012-09-07 | 2020-10-13 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10786566B2 (en) | 2012-09-07 | 2020-09-29 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9707293B2 (en) | 2012-09-07 | 2017-07-18 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10780163B2 (en) | 2012-09-07 | 2020-09-22 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9682145B2 (en) | 2012-09-07 | 2017-06-20 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10716853B2 (en) | 2012-09-07 | 2020-07-21 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9724414B2 (en) | 2012-09-07 | 2017-08-08 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US9731008B2 (en) | 2012-09-07 | 2017-08-15 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10688183B2 (en) | 2012-09-07 | 2020-06-23 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| US10772960B2 (en) | 2012-09-07 | 2020-09-15 | Coherus Biosciences, Inc. | Stable aqueous formulations of adalimumab |
| CN112656939A (en) * | 2014-09-22 | 2021-04-16 | 正大天晴药业集团股份有限公司 | Pharmaceutical composition of humanized antibody for vascular endothelial growth factor |
| CN112656939B (en) * | 2014-09-22 | 2023-12-08 | 正大天晴药业集团股份有限公司 | Pharmaceutical composition of humanized antibody for vascular endothelial growth factor |
| US11229702B1 (en) | 2015-10-28 | 2022-01-25 | Coherus Biosciences, Inc. | High concentration formulations of adalimumab |
| US11071782B2 (en) | 2016-04-20 | 2021-07-27 | Coherus Biosciences, Inc. | Method of filling a container with no headspace |
| US11576971B2 (en) | 2016-04-20 | 2023-02-14 | Coherus Biosciences, Inc. | Method of filling a container with no headspace |
| CN107619442B (en) * | 2016-07-15 | 2022-09-20 | 泰州迈博太科药业有限公司 | Recombinant anti-TNF-alpha fully human monoclonal antibody preparation |
| CN107619442A (en) * | 2016-07-15 | 2018-01-23 | 上海百迈博制药有限公司 | One kind recombinates the anti-full human monoclonal antibody preparations of TNF α |
| CN110151988A (en) * | 2018-02-11 | 2019-08-23 | 百奥泰生物制药股份有限公司 | A kind of human antibody preparation of targeted therapy TNF-α related disease |
| CN109239335A (en) * | 2018-09-11 | 2019-01-18 | 广州万孚生物技术股份有限公司 | Joint inspection test strips and preparation method thereof |
| CN116239678A (en) * | 2023-05-12 | 2023-06-09 | 广州悦洋生物技术有限公司 | Antioxidant composition, application, protective agent and solution |
| CN116239678B (en) * | 2023-05-12 | 2023-11-21 | 广州悦洋生物技术有限公司 | Antioxidant composition, application, protective agent and solution |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102988984B (en) | 2015-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102988984B (en) | Aqueous drug preparation of anti-TNF (tumor necrosis factor)-alpha human monoclonal antibody for strengthening stability | |
| EA027325B1 (en) | Etanercept formulations stabilized with xylitol | |
| Pochini et al. | Analysis of cytokine profile and growth factors in platelet-rich plasma obtained by open systems and commercial columns | |
| JPH01311027A (en) | Stabilization of human albumin solution of and obtained solution | |
| CN87104963A (en) | Stable pharmaceutical agent containing granulocyte colony stimulating factor and process for producing the same | |
| CA2532475A1 (en) | Formulation for a protein pharmaceutical without added human serum albumin (hsa) | |
| CN103160486A (en) | Preparation method of porcine thrombin | |
| TWI810196B (en) | Il-15 protein complex pharmaceutical composition and use thereof | |
| EP2676677A1 (en) | Highly concentrated anti-cd40 antibody pharmaceutical preparation | |
| CN103705914A (en) | Staphylococcus aureus vaccine and preparation method thereof | |
| CN101627967B (en) | Ambroxol hydrochloride liquid preparation and preparation method thereof | |
| CN102151257A (en) | Busulfan injection and preparation method thereof | |
| CN106729627B (en) | Recombinant human erythropoietin preparation | |
| RU2025120C1 (en) | Method of preparing of preparation containing factor g-csf stimulating the growth of granulocyte colonies | |
| JP2022500376A (en) | Compositions Containing Proteins and Polyalkoxy Fatty Acid Acyl Surfactants | |
| CN101991857B (en) | Stable pharmaceutical preparation and preparation method thereof | |
| JP2009539989A (en) | Formulation and production method of artesic acid for injection | |
| CN113521244B (en) | Argatroban injection and preparation method thereof | |
| CN101703466A (en) | Borneol injection and preparation method thereof | |
| WO2014032402A1 (en) | Erythropoietin recovery method | |
| CN103690949B (en) | Endomucin antibody medicament for prompting bone marrow regeneration | |
| JPH0424331B2 (en) | ||
| CN103877013A (en) | Asarin injection and preparation method thereof | |
| Ghawi et al. | Season variation and starvation period influence on the antithrombotic activity of leech saliva extract from the medicinal Malaysian leech, Hirudinaria manillensis | |
| CN111732672B (en) | Okra polygalacturonic acid with uric acid reducing effect and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20151216 Address after: 201203, room 1043, 602 Harley Road, Zhangjiang hi tech park, Shanghai, Pudong New Area Patentee after: Genor BioPharma Co., Ltd. Patentee after: Yuxi Jiahe Biotechnology Co., Ltd. Address before: 201203, room 1043, 602 Harley Road, Zhangjiang hi tech park, Shanghai, Pudong New Area Patentee before: Genor BioPharma Co., Ltd. |