CN104740612A - Liquid formulation of receptor antibody fusion protein - Google Patents

Abstract

The invention provides a liquid formulation of receptor antibody fusion protein. The invention relates to a liquid polypeptide preparation. The preparation contains a TNF:Fc polypeptide, an aggregation inhibitor, a buffering agent, a surfactant, a stabilizer, an osmotic pressure regulator and a chelating agent. The novel liquid formulation containing the TNF:Fc is stabilized and presents long-term stability at 4 DEG C and room temperature.

Description

A kind of liquid preparation of receptor antibody fusion rotein

Technical field

The present invention relates to a kind of aqueous formulation, the preparation method of said composition, medication and the therapeutic use that comprise TNFR:Fc.

Background technology

Tumor necrosis factor (TNF) is a kind of polypeptide cytokines relating to inflammation and acute phase response.TNF-α is present in the people suffering from rheumatoid arthritis or Crohn disease in a large number.The direct suppression being carried out TNF-α by biological preparation obtains marked improvement in the treatment of rheumatoid arthritis, and the extracellular of this proinflammatory cytokine suppresses to be confirmed to be a kind of effective treatment.A kind of such biological preparation is Embrel.

Embrel (TNFR:Fc) is a kind of dimer fusion protein, and this dimer fusion protein is made up of the extracellular ligand bound fraction of Fc people 75 kilodalton (p75) Tumor Necrosis Factor Receptors (TNFR) being partly connected to human IgG.The Fc component of Embrel is made up of CH2 domain, CH3 domain and hinge region, and CH1 domain lacks.It is produced by recombinant DNA technology in Chinese hamster ovary mammalian cell expression system.It is made up of 934 aminoacid, and has the apparent molecular weight of roughly 150 kilodaltons.

Usually, protein has very short half life, and after being exposed to various factors (as being not suitable for temperature, water-aerosphere face, high pressure, physical/mechanical stress, organic solvent and microbial contamination), experience degeneration (as assembled, dissociating and be adsorbed on vessel surface).Therefore, Denatured protein loses inherent physicochemical property and physiologically active.The degeneration of protein is normally irreversible, and therefore protein is once degeneration, and their natural character may can not return to original state.

In bio-pharmaceuticals industry, use the long term storage of the protein prepared of recombinant DNA technology in an aqueous formulation normally difficult task.In order to overcome the stability problem of the protein in aqueous formulation, prepare more stable human cytokines product by lyophilizing (lyophilization).Freeze-drying prods is attended by the sterile aqueous media for restoring usually.After recovery, these preparations typically have short effective storage life, even if be also like this when preserving under low temperature (such as, 5 DEG C).Commercially the example of the available TNF-alpha inhibitor in lyophilized form comprises with , and these two kinds of compositionss should be restored before the use.

The typical practice improving protein polypeptide stability can be solved by the concentration changing the key element in preparation, or changes said preparation by adding excipient.

US5580856 discloses and carrys out dry protein dry in stabilization formulations in case bioactive loss by adding a kind of stabilizing agent that restores when drying protein is rehydrated.Also describe a kind of for a kind ofly comprising the test kit producing a kind of preparation in the solvent of this recovery stabilizing agent by being dissolved in by this dry compositions at this.

US6171586 discloses a kind of stable aqueous pharmaceutical preparations, a kind of antibody that this pharmaceutical preparation comprises the treatment effective dose without prior lyophilizing, pH remained on about 4.5 to about 6.0 scope in a kind of buffer agent, a kind of surfactant and a kind of polyhydric alcohol, also disclose the purposes of this preparation simultaneously.

EP1314437 relates to a kind of invention of preparation of stabilisation, the preparation of this stabilisation contains a kind of antibody in glycine buffer and/or histidine buffer, additionally provide a kind of preparation method of preparation of proteinaceous stabilisation, these methods comprise and regulate pH with a kind of basic amino acid or a kind of basic amino acid derivant or their a kind of salt simultaneously.

EP1478394 relates to a kind of aqueous pharmaceutical compositions being suitable for long term storage of the polypeptide of the Fc domain containing a kind of immunoglobulin, manufacture method, medication and comprises the test kit of said composition.

Therefore, there is under making us desirably being provided in cold preservation the stability of enhancing and there is the liquid preparation of the TNFR:Fc of at least medium stability at nominal room temperature, thus avoiding the probability of inconvenience from reposition routine and mistake.

Summary of the invention

Pharmaceutical preparation of the present invention comprises a kind of polypeptide of purification, buffer agent, agglutination inhibitor, osmotic pressure regulator, stabilizing agent, surfactant, chelating agen and optionally antiseptic, and they are suitable combination.

For therapeutic use, monoclonal antibody or receptor antibody fusion rotein (as TNFR:Fc) use with high concentration, and the protein of known high concentration can increase gathering.In one aspect, in preparation compositions, need agglutination inhibitor making product remain on native state and be have bioactive, there is the shelf-life of prolongation and the condition of storage of enhancing.There is several aminoacid serving as agglutination inhibitor by increasing surface tension, preferential combination or hydration and preferentially interact.

Agglutination inhibitor reduces the trend of the formation aggregation of polypeptide.Aminoacid (as aspartic acid, phenylalanine, glutamic acid, alanine, histidine and lysine) reduces the gathering of the polypeptide containing Fc domain in preparation for a long time and is advantageous embodiments of the present invention.

In yet another aspect, need buffer agent to maintain the pH value of preparation.Buffer system of the present invention comprises phosphoric acid buffer agent, bicarbonate buffer agent, succinate buffer, acetate buffer, citrate buffer agent, amino acids and Tris buffer agent, they use individually or with suitable combination, provide desired from 5..5 to the pH value of 7.5 scopes.

In the present invention, surfactant is used for preventing TNFR:Fc to be adsorbed on the surface of bottle, ampoule bottle, cylindrical tubes (carpoule), cartridge case or syringe.Surfactant reduces the surface tension of protein solution, thus prevents its absorption or be gathered on a hydrophobic surface.The surfactant being applicable to invention formulation includes but not limited to Polysorbate (such as polysorbate 20 or 80); Pa Luoshamu (such as Pa Luoshamu 188 or 168); Sorbitan ester and derivant thereof; Triton; Sodium lauryl sulphate; OG sodium; Dodecyl, myristyl, sub-oleoyl-or stearoyl-sulfobetaines; Dodecyl, myristyl, sub-oleoyl or stearoyl-sarcosine; Sub-oleoyl _, myristyl or cetyl-betanin; Dodecanamide propyl _, cocamidopropyl propyl amide _, sub-oleamide propyl group _, lima bean fringed pink amido propyl _, palmitamide propyl group-or isostearoyl amine CAB (such as dodecanamide propyl); Lima bean fringed pink amido propyl _, palmitamide propyl group-or isostearoyl amine propyl group-dimethylamine; Sodium methyl cocoyl taurate or the horizontal acid disodium of methyl oleoyl cattle; And M0NAQUAT ^tMseries (Mona Industries, Inc., Paterson, N.J.); Polyethylene Glycol, the copolymer (such as, Pluronics, PF68 etc.) of poly-propanol and ethylene glycol and propylene glycol.They individually or combinationally use.

In one embodiment, the stabilizing agent used in the present invention is selected from lower group, and this group is made up of the following: amino acids, as glycine, arginine, alanine, lysine, proline, serine, etc. and its their salt, they are individually or combinationally use; Monosaccharide, as glucose and mannose and their analog, they are individually or combinationally use; Disaccharides, as sucrose, trehalose, maltose and their analog, they are individually or combinationally use; Sugar alcohols, as mannitol, Sorbitol and their analog, they are individually or combinationally use; And polysaccharide, as glucosan, Polyethylene Glycol and their analog, they are individually or combinationally use.

Osmotic pressure regulator is understood to be a kind of molecule facilitating the osmolarity of solution.Preferably, a kind of osmolarity of pharmaceutical composition is conditioned to improve the stability of active component to greatest extent, also the discomfort of patient during administration is reduced to bottom line simultaneously.The embodiment being applicable to the osmotic pressure regulator changing osmolarity comprises, but be not limited to, aminoacid (arginine, cysteine, histidine, etc.), salt (sodium chloride, potassium chloride, calcium chloride etc.) and/or saccharide (sucrose, glucose, mannitol and their analog).

Embrel tends to fragmentation in storage process, so need some fragmentation inhibitor to carry out the optimal formulation compositions of manufacturing needles to Embrel.The known influential EDTA of fragmentation to preventing in many products (as interferon, alemtuzumab, etc.) is used in this preparation.

Chelating agen is stable or prevent free metal ion and interested proteins react.The example that may be used for the chelating agen in the present invention comprises, but be not limited to, EDTA (ethylenediaminetetraacetic acid), HEDTA (hydroxyethylethylene diamine tri-acetic acid), NTA (nitrilotriacetic acid(NTA)), DTPA (second diene pentaacetic acid) and citric acid.

Antiseptic refers to and adds in preparation as a kind of compositions of antibacterial or material.Preferably, the preparation of the TNFR:Fc containing preservation of the present invention meets the legal of preservative effectiveness or supervision criterion, using the multipurpose product as a kind of viable commercial, preferably for the mankind.The antiseptic used in the present invention is selected from lower group, this group is made up of the following: phenol, metacresol, paracresol, orthoresol, chlorocresol, alkyl parabens (methyl parahydroxybenzoate, ethylparaben, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, etc.), benzethonium chloride, dehydro sodium acetate or thimerosal, they are individually or combinationally use.But in the present invention, the use of antiseptic is optional, and be preferred when designing multi-dose formulation.

The invention provides a kind of liquid preparation comprising the novelty of TNFR:Fc, it presents long-time stability under 4 DEG C and room temperature.

The invention discloses a kind of pharmaceutical preparation, comprise the polypeptide of a kind of TNF:Fc, agglutination inhibitor, buffer agent, surfactant, stabilizing agent, osmotic pressure regulator and chelating agen.

Said medicine preparation, wherein this agglutination inhibitor is selected from lower group, and this group is made up of the following: aspartic acid, phenylalanine, glutamic acid, alanine, glycine, histidine and lysine.

Said medicine preparation, wherein this buffer agent is selected from lower group, and this group is made up of the following: sodium phosphate, histidine, potassium phosphate, sodium citrate or potassium citrate, maleic acid, ammonium acetate, Tris (tris), acetate and diethanolamine or their combination.

Said medicine preparation, wherein this surfactant is selected from lower group, and this group is made up of the following: the nonionic surfactant based on Polysorbate, the nonionic surfactant based on Pa Luoshamu or their combination.

Said medicine preparation, wherein this stabilizing agent is selected from lower group, sucrose, trehalose, maltose, mannitol, and glycine, arginine, alanine, lysine, proline and serine and their salt composition, individually or combinationally use.

Said medicine preparation, wherein this osmotic pressure regulator is selected from lower group, and this group is made up of the following: sodium chloride, potassium chloride, sodium sulfate or their combination.

Said medicine preparation, wherein this chelating agen is selected from lower group, and this group is made up of the following: EDTA (ethylenediaminetetraacetic acid), HEDTA (hydroxyethylethylene diamine tri-acetic acid), NTA (nitrilotriacetic acid(NTA)), DTPA (second diene pentaacetic acid) and citric acid or their combination.

In one embodiment, the pharmaceutical preparation of the preferred a kind of TNFR:Fc of the present invention, comprise TNFR:Fc fusion rotein, aspartic acid, phosphate buffer, sodium chloride, sucrose, arginine salt, Pa Luoshamu and disodiumedetate, wherein TNFR:Fc exists with the concentration of 10mg/ml to 100mg/ml.

In one embodiment, the pharmaceutical preparation of the preferred a kind of TNFR:Fc of the present invention, comprise the Embrel of 25mg/ml to 50mg/ml, the aspartic acid of 1mM to 50mM, the sodium phosphate buffer of 1OmM to 50mM, the sucrose of 10mg/ml to 50mg/ml, the arginine monohydrochloride of 1mM to 50mM, the NaCl of 50mM to 150mM, the EDTA of 0.1mM to 2mM, the Pa Luoshamu 188, pH of 0.1mg/ml to 1Omg/ml be 6.O to 7.O.

Pharmaceutical preparation disclosed by the invention, uses by intravenous injection or subcutaneous injection, is used for the treatment of as inflammatory diseasess such as rheumatoid arthritis, ankylosing spondylitis, psoriasises.

The novelty of TNFR:Fc described in the present invention, heat-resisting aqueous formulation has the following advantages:

1. relate to the use of agglutination inhibitor, prevent the gathering of fusion rotein in prolonged storage.

2. relate to a kind of use of stabilizing agent, prevent undesirable cracking in prolonged storage.

3. for this aqueous formulation provides better stability to maintain its activity for prolonged period of time, thus ensure the rational shelf-life.

Even if 4. at high temperature also for this aqueous formulation provides better stability.

Following instance illustrates the pharmaceutical composition that describes in the present invention and carries out the present invention to obtain the mode of the aqueous pharmaceutical preparations of stable TNFR:Fc.These examples never should be interpreted as the restriction to scope of the present invention.

Accompanying drawing explanation

Fig. 1 shows the comparison transition mid point of several formulations.

Detailed description of the invention

The Selection and screening of embodiment 1 agglutination inhibitor

Agglutination inhibitor is screened at 40 DEG C.

Have studied the Embrel preparation with different agglutination inhibitors (as lysine, aspartic acid, glycine and histidine).These preparations are also containing other the excipient according to details given in Table 1.

Table 1: the qualification of agglutination inhibitor

Sample formulation hatches one month at 40 DEG C, is then analyzed by SDS-PAGE.SDS-PAGE is used as a kind of analytical technology, can isolate free with kind that is high molecular according to molecular weight from native protein.Owing to having high gathering tendency when the monoclonal antibody of high concentration and fusion rotein, non-reduced SDS-PAGE is used to evaluation covalency aggregation.Owing to there is many disulfide bond in fusion rotein and monoclonal antibody, under reduction SDS-PAGE, check the fragmentation of protein.

SDS-PAGE analysis result finds, just to assemble and with regard to fragmentation, F2 and F3 has more degraded, but just with regard to the viewed aggregation of F1 and F4 and fragment, does not have gross differences.Owing to not observing the significant difference of impurity at this temperature, studying these two preparations (F1 and F4) at a higher temperature further increases degradation kinetics and degradation product.

The screening of agglutination inhibitor at 48 DEG C and 55 DEG C.

In order to find out best agglutination inhibitor between aspartic acid (F1) and histidine (F4), make these two preparations all charged at 48 DEG C and 55 DEG C.Result shows, compared with aspartic acid (F1), histidine (F4) under two kinds of probe temperatures (namely at 48 DEG C 36h and at 55 DEG C 24h) just assemble and demonstrate higher degraded with regard to fragmentation.Aspartic acid is proved to be at all temperatures for preventing gathering and fragmentation from being effective.

The effect of embodiment 2EDTA

Checked the effect of EDTA in F4 preparation (continuing 5 days at 40 DEG C), and analyzed by SE-HPLC, with size exclusion chromatography (SEC) isolated protein and relative substance (sizes based on them) thereof.This technology for detecting the gathering of Embrel and fragmentation is useful, and provides in result table 2 below.SDS PAGE is used to compare aggregation and clip types equally.

Table 2: the SE-HPLC result demonstrating the preparation of EDTA impact

Especially, with regard to fragment, according to SE-HPLC result, compared with the preparation not having EDTA, find that F4 has lower degradation rate.

By SDS PAGE, find that F4 has the low-molecular-weight band of small amount after sample exposes 5 days at 40 DEG C.Therefore, EDTA plays an important role in the fragmentation preventing Embrel.

The effect of embodiment 3 buffer agent

As the Embrel preparation with phosphate buffer and histidine buffer that provides in table 3 (F1, F5, F6 and F7) at 50 DEG C charged 2 days, then analyzed by SE-HPLC and differential scanning calorimetry (DSC).DSC is a kind of technology of the thermodynamic stability for measuring protein.Embrel has three transition, Tm1 corresponding to TNFR, Tm2 corresponding to Fc part CH2 domain and Tm3 corresponding to Fc part CH3 domain (higher Tm represents higher stability).

Table 3: the qualification of buffering component

Table 4: the SE-HPLC result of buffering component qualification

According to SE-HPLC result, compared with the preparation (F6, F7) containing histidine buffer, the compositions (F1, F5) with the phosphate buffer as buffer agent has high-purity and less aggregation.According to dsc analysis, the transition mid point at the first and second transition peaks of phosphate buffer is respectively than histidine buffer agent formulation high about 1 DEG C and 2 DEG C respectively.The mid point at the 3rd transition peak in two kinds of buffer agents is almost similar (Fig. 1).

Due to compared with histidine buffer, observe phosphate buffer have lower degradation rate (as SE-HPLC result prove) and high thermodynamic stability (being proved by dsc analysis), phosphate buffer is by preferably as the buffer system be applicable to.

Meanwhile, we also find, increase L-arginine hydrochlorate and sucrose jointly as stabilizing agent in F5 and F7, greatly can improve preparation purity and reduce aggregation to be formed.

The qualification of embodiment 4 surfactant

Optimal material standed in stability is filtered out from non-ionic surface active agent Polysorbate (such as polysorbate 20 or 80) and Pa Luoshamu (such as Pa Luoshamu 188 or 168).

For the stability of desired protein, employ polysorbate 20 and Pa Luoshamu 188.Gathering in storage process and fragmentation has been evaluated by SE-HPLC method.Report the test is in table 6 and table 7.

Table 5: formulation ingredients table

Embrel	50mg	50mg
			Sucrose	20mg	10mg
L-arginine hydrochlorate		25mM
			Phosphate buffer	25mM	25mM

Aspartic acid	20mM	20mM
			Sodium chloride	100mM	100mM
EDETATE SODIUM	1mM	1mM
			Surfactant	Specifically in table 6	Specifically in table 7

Table 6: the surfactant of varying level and type is on containing the impact of sucrose as stabilizing agent

Table 7: the surfactant of varying level and type is on containing the impact as stabilizing agent of sucrose and L-arginine hydrochlorate

According to SE-HPLC result, compared with the preparation containing polysorbate 20 surfactant, the preparation compositions had as the Pa Luoshamu 188 of surfactant has higher purity and less aggregation.

Meanwhile, we also find, increase L-arginine hydrochlorate and sucrose, jointly as stabilizing agent, greatly can improve preparation purity and reduce aggregation formation.

The preparation of embodiment 5TNFR:Fc fusion protein formulations

Except Pa Luoshamu 188, all components provided in table 8 is all dissolved in the water of 80%, and they is mixed fully by continuous stirring.In this solution, add Pa Luoshamu 188, make ultimate density be 0.8mg/ml.With water, the volume of formulation buffer agent is transferred to 90%.With any applicable acid/alkali, pH value is adjusted to 6.3, and with water, volume is supplied to 100%.Said preparation buffer agent is filtered, and with this filtered formulation buffer agent by the concentration needed for Embrel volume dilution one-tenth.Store 1,3 and 6 months at-20 DEG C, 5 ± 3 DEG C and 25 ± 2 DEG C after, analyze Embrel preparation in the zero-time by SE-HPLC, SDS-PAGE and HIC.These report the tests are in table 9.

As mentioned above, pharmaceutical preparation of the present invention is prepared by adding agglutination inhibitor to the polypeptide of purification.In addition, a kind of buffer agent, osmotic pressure regulator, surfactant, stabilizing agent, fragmentation inhibitor and a kind of other excipient etc. can be added if desired.Any those of ordinary skill in the art will be understood that, the combination of the various components be included in the composition can have been come with any order suitably, namely, first can add, add in centre or in the end add buffer agent, and also can first add, to add in centre or in the end to add osmotic pressure regulator etc.Equally, those of ordinary skill in the art will be understood that, some in some combination in these chemical substances are inconsistent, and therefore, they easily can be had similar quality but be that compatible different chemical substance replaces in relative mixtures.

Table 8

Composition	Concentration
		Embrel	50mg/ml
Sucrose	10mg/ml
		L-arginine hydrochlorate	5.3mg/mL
One water sodium hydrogen phosphate	2.6mg/mL
		Disodium hydrogen phosphate,anhydrous	0.9mg/mL
Aspartic acid	2.6mg/mL
		Sodium chloride	5.8mg/mL
EDETATE SODIUM	0.37mg/mL
		Pa Luoshamu 188	0.8mg/mL

The stability test of TNFR:Fc aqueous formulation, these biological tests carry out according to the specification of European Pharmacopoeia.

SDS PAGE[assembles (non-reduced) and fragmentation (reduction)]

Owing to having high gathering tendency when the monoclonal antibody of high concentration and fusion rotein, non-reduced SDS-PAGE is used to evaluation covalency aggregation.Due to TNFR by disulfide-bonded to the hinge region of Fc and because there is many disulfide bond in Fc and TNFR, under reduction SDS PAGE, check the fragmentation of protein.

SE-HPLC (assembling and fragmentation)

With size exclusion chromatography (SEC) isolated protein and relative substance (sizes based on them) thereof, above-mentioned technology is for detecting the gathering of Embrel and fragmentation is useful.

HIC (gathering, fragmentation, truncate and misfolding)

HIC carrys out isolated protein based on hydrophobicity.Peak I in HIC determines fragment, and peak 2 is corresponding to the active dimeric of Embrel, and peak 3 is corresponding to aggregation.The leading peak I observed in some cases is corresponding to truncate impurity.

Differential scanning calorimetry (DSC)

DSC is a kind of technology of the thermodynamic stability being used for measuring protein.Embrel has three transition, Tm I corresponding to TNFR, Tm2 corresponding to Fc part CH2 domain and Tm3 corresponding to Fc part CH3 domain (higher Tm determines higher stability).

About aqueous formulation stability in be collected in table 9.

Table 9: the stability test result (-20 DEG C, 5 ± 3 DEG C, 25 ± 2 DEG C) of Embrel preparation

(a: each temperature measuring 3 groups of preparations are averaged.）

The stability test of embodiment 6 TNFR:Fc liquid preparation at 40 DEG C

Prepare the liquid preparation comprising the Embrel with the composition provided in table 8.The solution of preparation stores 1 month by O.2 μm filter sterilised under lamina air flow at 40 DEG C.Analyze said preparation by SE-HPLC, SDS-PAGE and HIC store zero-time, 7 days, 15 days, 21 days at 40 DEG C after and check its biological activity.Report the test is in table 10.

Table 10: the stability test result (40 DEG C) of Embrel preparation

Measure a	Zero-time	7 days	15 days	21 days	30 days
						The gathering that SE-HPLC measures	1%	1.4%	2.0%	3.0%	4.7%
The fragmentation that SE-HPLC measures	0%	0.1%	0.4%	0.6%	1.0%
						HIC peak 1	0%	NA	NA	NA	2.0%
HIC peak 2	95%	NA	NA	NA	92.2%
						HIC peak 3	5%	NA	NA	NA	5.8%

(a: each temperature measuring 3 groups of preparations are averaged.）

The stability test of embodiment 7 TNFR:Fc liquid preparation at 50 DEG C

Prepare the liquid preparation comprising the Embrel with the composition provided in table 8.The solution of preparation stores 3 days by O.2 μm filter sterilised under lamina air flow at 50 DEG C.Said preparation is analyzed by SE-HPLC, SDS-PAGE and HIC after storing 2 days and 3 days in the zero-time and at 50 DEG C.Report the test is in table 11.

Table 11: the result (50 DEG C) of the stability test of Embrel preparation

Measure a	Zero-time	2 days	3 days
				The gathering that SE-HPLC measures	1%	5.4%	10.8%

The fragmentation that SE-HPLC measures	0%	0%	0%
				HIC peak 1	0%	0%	0%
HIC peak 2	95%	94.2%	91.6%
				HIC peak 3	5%	5.8%	8.4%

(a: each temperature measuring 3 groups of preparations are averaged.）

The combination of applicable buffer agent, agglutination inhibitor, osmotic pressure regulator, stabilizing agent, surfactant, chelating agen (and optionally having antiseptic in their suitable combination) is used to prepare the fusion protein formulations of these novelties.

Included the biological activity TNFR:Fc of effective amount by the preparation of described invention preparation, it is used to the inflammatory diseases for the treatment of people, as rheumatoid arthritis, ankylosing spondylitis, psoriasis etc.They are used preferably as injection aqueous solution, can be diluted to certain density aqueous solution intravenous injection as required and use, or preferred filling is used by subcutaneous injection in precharging type syringe.

Claims (10)

1. a pharmaceutical preparation, comprises the polypeptide of a kind of TNF:Fc, agglutination inhibitor, buffer agent, surfactant, stabilizing agent, osmotic pressure regulator and chelating agen.

2. pharmaceutical preparation as claimed in claim 1, wherein this agglutination inhibitor is selected from lower group, and this group is made up of the following: aspartic acid, phenylalanine, glutamic acid, alanine, glycine, histidine and lysine.

3. pharmaceutical preparation as claimed in claim 1, wherein this buffer agent is selected from lower group, and this group is made up of the following: sodium phosphate, histidine, potassium phosphate, sodium citrate or potassium citrate, maleic acid, ammonium acetate, Tris (tris), acetate and diethanolamine or their combination.

4. pharmaceutical preparation as claimed in claim 1, wherein this surfactant is selected from lower group, and this group is made up of the following: the nonionic surfactant based on Polysorbate, the nonionic surfactant based on Pa Luoshamu or their combination.

5. pharmaceutical preparation as claimed in claim 1, wherein this stabilizing agent is selected from lower group, sucrose, trehalose, maltose, mannitol, and glycine, arginine, alanine, lysine, proline and serine and their salt composition, individually or combinationally use.

6. pharmaceutical preparation as claimed in claim 1, wherein this osmotic pressure regulator is selected from lower group, and this group is made up of the following: sodium chloride, potassium chloride, sodium sulfate or their combination.

7. pharmaceutical preparation as claimed in claim 1, wherein this chelating agen is selected from lower group, and this group is made up of the following: EDTA (ethylenediaminetetraacetic acid), HEDTA (hydroxyethylethylene diamine tri-acetic acid), NTA (nitrilotriacetic acid(NTA)), DTPA (second diene pentaacetic acid) and citric acid or their combination.

8. the pharmaceutical preparation of a TNFR:Fc, comprise TNFR:Fc fusion rotein, aspartic acid, phosphate buffer, sodium chloride, sucrose, arginine salt, Pa Luoshamu and disodiumedetate, wherein TNFR:Fc exists with the concentration of 10mg/ml to 100mg/ml.

9. pharmaceutical preparation as claimed in claim 8, comprise the Embrel of 25mg/ml to 50mg/ml, the aspartic acid of 1mM to 50mM, the sodium phosphate buffer of 1OmM to 50mM, the sucrose of 10mg/ml to 50mg/ml, the arginine monohydrochloride of 1mM to 50mM, the NaCl of 50mM to 150mM, the EDTA of 0.1mM to 2mM, the Pa Luoshamu 188, pH of 0.1mg/ml to 1Omg/ml be 6.O to 7.O.

10. the pharmaceutical preparation as described in claim 1-10, uses by intravenous injection or subcutaneous injection, is used for the treatment of as inflammatory diseasess such as rheumatoid arthritis, ankylosing spondylitis, psoriasises.