CN101797383A - Hydro-acupuncture preparation of monoclonal antibody - Google Patents

Abstract

The invention provides a preparation method of a hydro-acupuncture preparation of an anti-EGFR-antibody. The hydro-acupuncture preparation of the anti-EGFR-antibody consists of anti-EGFR-antibody albumen, a buffer salt and a surface active agent, and has the advantages that the buffer salt not only has the effect of buffering pH, but also exists in solution as a protective agent of the albumen; and simultaneously, PEG replaces hemolytic Tween as the surface active agent, so that a formula is safe. The hydro-acupuncture preparation for the anti-EGFR-antibody albumen has the advantages of high stability, prolonged valid period and low cost.

Description

A kind of hydro-acupuncture preparation of monoclonal antibody

Technical field

The present invention relates to a kind of hydro-acupuncture preparation of anti-EGFR monoclonal antibodies.

Background technology

EGF-R ELISA (epidermal growth factor receptor, EGFR) in growth, reparation and existence, the new vessels of regulate tumor cell generate, attack and shift, have important effect, expression is all arranged in the human tumor of quite a few simultaneously.In addition, in a lot of malignant tumor, the expression of the EGFR often back and lower survival rate with relatively poor is relevant.Therefore can infer that if there is medicine can block the activity of EGFR, that will suppress its phosphorylation and signal conduction, thereby plays the antitumor action of multiple approach, also can increase the antitumor curative effect of chemotherapy and radiation simultaneously.Compare with natural part, when the monoclonal antibody of anti-epidermal growth factor receptor (EGFR) combines with EGFR, have higher affinity, the cell pathway of therefore blocking the EGFR mediation is a kind of high specific model of action that has.

The monoclonal antibody of anti-EGFR is aimed at the target antibody of extracellular domain EFGR, and clinical practice has shown good antineoplastic activity, and does not produce serious side reaction.The anticancer principle of anti-EGFR monoclonal antibodies is just based on this.

Existing anti-EGFR-antibodies preparation patent:

Germany S Ma Sesi and H-C bridle obtain the solid form preparation (application number/patent No.: 200480034587) of anti-EGFR-antibodies by precipitation reagent and crystal.Its shortcoming is that the albumen that precipitation reagent obtains can't redissolve sometimes.

The H-C bridle of Merck Patent GmbH; The R Miller; U martini-Ma Er; The U Haas; The C Ingeborg Bachmann is invented the stable liquid pharmaceutical formulation that comprises Cetuximab (Cetuximab), and (application number: 02814105.9), described Cetuximab is the chimeric mAb of a kind of anti-epidermal growth factor receptor (EGF receptor).Institute's detection method:

Purity detecting: high performance liquid chromatography, pillar TSK3000SW _XLMobile phase: 20mmol phosphate buffer 0.4mol/LNa ₂SO ₄

Isoelectric point, IP: according to " Chinese pharmacopoeia point focusing electrophoresis method such as version in 2005.

Endotoxin detects: according to " Chinese pharmacopoeia version detection of bacterial endotoxin in 2005 method.

Outward appearance: according to " Chinese pharmacopoeia version visible foreign matters in 2005 inspection technique.

Active: according to the A418 detection method with commercially available Erbitux (Merck ﹠ Co., Inc.) as reference substance.Setting its activity is 100%, and liquid drugs injection comparative solution, solution A, solution B adopt fiducial value active with it.

Accelerated stability experimental result (25 ℃)

The result shows according to stability test: the preparation stored stability that the buffered with amino acid system is formed obviously is better than the preparation that phosphate system is formed.

The specific embodiment

By the following examples, experimental example describes content of the present invention in detail, but these embodiment are not construed as limiting the invention.

Embodiment 1: the liquid drugs injection comparative solution

Composition	Concentration
		Cetuximab	??100mg
Sodium dihydrogen phosphate	??20mg
		Sodium hydrogen phosphate	??66mg
Sodium chloride	??424mg
		Water for injection	Add to 50ml
PH value	??7.1

Embodiment 2: solution A

Composition	Concentration
		Cetuximab	??300mg
Arginine	??40mmol/L
		Histidine	??40mmol/L
??PEG6000	??1％
		Water for injection	Add to 10ml
PH value	??7.1

Embodiment 3:

Study on the stability, the bin stability of the above 3 kinds of prescriptions of employing medicine accelerated stability The effects, 25 ℃ of temperature.

State the storage-stable enhancing of preparation and can be used for the treatment of tumor through the parenteral approach.S Ma Sesi and H-C bridle prepare the highly concentrated liquid of anti-EGFR-antibodies again by ultrafiltration, invented the method (application number/patent No.: 200580004883) of anti-EGFR-antibodies liquid preparation.These two inventions have largely been alleviated in the past because of the low inconvenience that brings of protein drug concentration; but weak point is arranged also; at first its patent is a buffer system with salt such as phosphate or acetate; the albumen that the form of liquid preparation exists will produce opalescence or precipitation when salinity is high slightly; physical and chemical reaction takes place easily; thereby lose activity, secondly when albumen excessive concentration in aqueous solution has the protective agent protection again, do not assemble easily, easily produce immunoreation behind the injection human body.

Summary of the invention

That anti-EGFR-monoclonal antibody comprises is the Mus source, human mouse chimeric, the anti-EGFR-monoclonal antibody of full-length human.

Pharmaceutical formulation of the present invention is simple, and the albumen system is stable.Be convenient to large-scale production, store and transportation.The system of forming with acidic amino acid and basic amino acid has very strong buffer capacity and to proteic protective capability.There is not the unacceptable reagent of pharmacology in this pharmaceutical formulation, thereby has good safety and toleration.

The present invention is a kind of hydro-acupuncture preparation that contains anti-EGFR-monoclonal antibody, and the composition of preparation comprises anti-EGFR-monoclonal antibody, buffer salt, surfactant.

Anti-EGFR-monoclonal antibody hydro-acupuncture preparation protein content is 2-100mg/mL, more preferably 5-75mg/mL.The protein formulation of this high concentration can alleviate the big misery of patient's medication injection volume greatly.

Buffer system of the present invention comprises the acceptable buffer system of human body, more preferably arginine, histidine system and arginine, glycine system.Its concentration is 0-200mM, more preferably 10mM-100mM.The buffer pH scope is between 6.0-9.0.More preferably pH6.5-7.5.

Surfactant of the present invention is the PEG class, more preferably PEG400, and PEG4000, PEG6000, surfactant concentration are 0.001%-5%, more preferably 0.005%-4%.

Claims (7)

1. hydro-acupuncture preparation that contains anti-EGFR-antibody, it is characterized in that in pharmaceutical formulation, utilizing acidic amino acid and basic amino acid promptly as buffer system again as protective agent, utilize PEG to replace tween as surfactant.

2. according to claim 1, it is characterized in that anti-EGFR-antibody content is 1-100mg/mL, more preferably 2-60mg/mL.

3. according to claim 1, it is characterized in that buffer salt system comprises the acceptable buffer system of human body, more preferably arginine, histidine system and arginine, glycine system.

4. according to claim 3, the concentration that it is characterized in that buffer system is 0-200mM, more preferably 10mM-100mM.

5. according to claim 3, the buffer pH scope that it is characterized in that buffer system is between 6.0-9.0.More preferably pH6.5-7.5.

6. according to claim 1, it is characterized in that surfactant is the PEG class, more preferably PEG400, PEG4000, PEG6000.

7. according to claim 6, it is characterized in that surfactant concentration is 0.001%-10%, more preferably 0.005%-4%.