Field of Invention
The invention provides stable pharmaceutical compositions comprising tumor necrosis
factor receptor Fc fusion protein (TNFR:Fc). The invention also provides methods of
manufacturing the composition, method of administration and kits containing the same.
Background of Invention
Tumor necrosis factor (TNF) alpha is a cytokine that promotes the inflammation and its
associated signs by binding to its receptor. It is produced by macrophages and many other
immune cells. It is involved in pathogenesis of many inflammatory disorders like
rheumatoid arthritis, psoritic arthritis, SLE, Crohn's disease etc. Hohmann et al
(Hohmann et al. 1989 J Biol Chem. 25, 14927-34) identified 2 distinct receptors of TNFalpha
which are present on different cell types viz. myeloid cells and epithelial cells.
Using monoclonal antibodies, Brockhaus et al (Brockhaus et al. 1990 ProcNatlAcadSci U
S A., 87(8), 3127-31) demonstrated that both TNF-alpha and beta bind to both the
receptors with high affinity.
Tumor necrosis factor-alpha (TNF-alpha) is a central regulator of inflammation, and
TNF-alpha antagonists may be effective in treating inflammatory disorders in which
TNF-alpha plays an important pathogenetic role. Inhibition of TNF has proven to be an
effective therapy for patients with rheumatoid arthritis and other forms of inflammatory
disease including psoriasis, psoriatic arthritis, and ankylosing spondylitis, inflammatory
bowel disease. One such TNF-alpha antagonist is Etanercept.
Etanercept is a dimeric fusion protein produced by recombinant DNA technology where
gene of soluble, ligand binding portion of TNF receptor 2 is fused with gene of Fc
component of human IgGl to give the desired fusion protein (US 7648702). Etanercept is
expressed in CHO cells. The Fc component of Etanercept lacks CHI domain but has
CH2, CH3 domains and hinge region. The fusion protein has approximate molecular
weight of 150 kD and consists of 934 amino acids. Etanercept interferes with TNF and
acts as a TNF inhibitor due to which it can be used as a biopharmaceutical to treat
autoimmune diseases. It prevents progressive destruction of joints in patients with
rheumatoid arthritis and the arthritis of psoriasis.
Due to its unique structure, Etanercept binds 50-100 folds more efficiently to TNF alpha
than its endogenous receptor (Gofeeet al. 2003/ Am AcadDermatol. 49, S105-111,
Strober 2005 SeminCutan Med Surg. 24; 28-36). Additionally, due to its dimeric nature it
can bind to 2 TNF alpha molecules as compared to one bound by endogenous receptor.
Conjugation of this molecule to Fc region of IgG increases the half life as compared to
endogenous soluble form. Commercially, Etanercept is available in both Lyophilized and
liquid forms.
The most important feature of a composition is to help the protein to retain its structural
conformation or its activity. The stability of protein in a composition can be related with
long-term storage. It is understood to mean that the active polypeptide of the
pharmaceutical composition does not substantially lose its activity as compared to the
composition at the beginning of storage.
All polypeptides have an Isoelectric Point (pi), which is generally defined as the pH at
which a polypeptide carries no net charge. It is known in the art that protein solubility is
typically lowest when the pH of the solution is equal to the isoelectric point (pi) of the
protein.
The Tm of the Fab domain of a protein is a good indicator of the thermal stability of a
protein and may further provide an indication of the shelf-life. Tm values of proteins
determined by differential scanning calorimetry, give insight into heat-induced changes in
protein conformation, mechanisms of protein unfolding and stabilization in solution. A
lower Tm indicates less stability of a protein in given solution, whereas a higher Tm
indicates a better stability of the protein. The Tm of the protein will vary based on the
formulation composition which in turn reflects its stability in respective formulation.
During long term storage, both aqueous and lyophilized compositions of proteins can lose
active protein due to aggregation or degradation. Aggregation of the protein can lead to
immunogenicity and is undesirable. Since the concentration of Etanercept used in the
composition is high, there is a likely possibility of protein aggregation during long term
storage. To improve the stability of the protein either the concentration of the existing
excipients can be varied or new excipients can be added to modify the composition.
US Patent Nos US 5,215,743; US 7,648,702; US application US 20070053906 and WO
2011141926 disclose pharmaceutical compositions comprising aqueous composition of
TNF-binding protein comprising a TNF-binding protein, a buffer and an isotonicity agent.
Summary of the Invention
In an embodiment, the invention is related to a stable pharmaceutical composition
comprising TNFR:Fc fusion protein and phosphate - citrate buffer.
In another embodiment, the invention is related to a stable pharmaceutical composition
comprising TNFR:Fc fusion protein, phosphate - citrate buffer and anti-aggregating agent
selected from L-glycine, urea and 2-hydroxypropyl beta-cyclodextrin (HPBCD).
In another embodiment, the invention is related to a stable pharmaceutical composition
comprising TNFR:Fc fusion protein, phosphate - citrate buffer, anti-aggregating agent
selected from L-glycine, urea and HPBCD, a tonicity modifying agent and a stabilizing
agent.
In yet another embodiment, the invention is related to the method of treating a disease
using the stable pharmaceutical composition of the present invention. The disease may be
rheumatoid arthritis, polyarticular juvenile idiopathic arthritis, psoriatic arthritis,
ankylosing spondylitis or plaque psoriasis.
In another embodiment the invention is related to a kit or container containing the
pharmaceutical composition of the invention.
The details of one or more embodiments of the invention set forth below are illustrative
only and not intended to limit to the scope of the invention. Other features, objects and
advantages of the inventions will be apparent from the description and claims.
Detail Description of Invention
The invention provides a stable pharmaceutical composition comprising TNFR molecules
fused to an Fc portion of a human immunoglobulin (TNFR:Fc fusion protein). More
particularly, the invention relates to the stable pharmaceutical composition of etanercept
in phosphate - citrate buffer, which displays a lower degradation potential.
In an embodiment of the invention, the TNFR:Fc fusion protein is etanercept.
It has been reported in US 7648702 patent and WO201 1141926 application that the
pharmaceutical compositions of Etanercept using L-glycine as anti-aggregating agent in
phosphate buffer are not stable as compared to the compositions of etanercept with other
amino acid such as arginine, proline, lysine, aspartic acid as anti-aggregating agent in
phosphate buffer. The WO201 1141926 application discloses that the composition
comprising Etanercept in phosphate buffer and L-glycine as anti-aggregating agent
showed aggregation as well as fragmentation products.
While studying the Etanercept compositions in different buffers and using different antiaggregating
agents it was observed that Etanercept composition comprising phosphate -
citrate buffer with L-glycine as anti-aggregating agent showed improved stability as
compared to Etanercept composition comprising phosphate buffer with L-glycine as antiaggregating
agent at 5 °C and 40 °C.
As illustrated in the example section, the stability of Etanercept composition essentially
consisting of phosphate - citrate buffer in combination with L-glycine as anti-aggregating
agent were assessed during a 6 months stability study at 5 °C as well as 2 weeks study at
40 °C (stress conditions stability studies). Compositions comprising the phosphate -
citrate buffer system were determined to be superior as compared to composition
comprising phosphate buffer system with respect to the % aggregation products and &
degradation products as determined by SEC.
The stable pharmaceutical composition used herein means that the TNFR:Fc fusion
protein exhibits following features:
i . The stable pharmaceutical composition of TNFR:Fc fusion protein in phosphate -
citrate buffer exhibits improved stability as compared to the composition of
etanercept comprising phosphate buffer, arginine and sodium chloride. The %
aggregates are less in the Etanercept composition comprising L-glycine as antiaggregating
agent in phosphate-citrate buffer as determined after 2 weeks of
storage at 40°C by SEC.
ii. The stable pharmaceutical composition of TNFR:Fc fusion protein in phosphate -
citrate buffer exhibits less than 5% high and low molecular weight impurities
similar to the innovator composition of etanercept comprising phosphate buffer,
arginine and sodium chloride after 2 weeks of storage at 40°C by SEC.
iii. The stable pharmaceutical composition of TNFR:Fc fusion protein in phosphate -
citrate buffer, glycine exhibits approximately 8% high and low molecular weight
impurities. Whereas the composition of etanercept comprising phosphate buffer,
glycine as an anti-aggregating agent showed 15% impurities, after 2 weeks of
storage at 40°C by SEC.
In another embodiment the invention relates to the pharmaceutical composition of
Etanercept in phosphate -citrate buffer with other anti-aggregating agents such as urea,
HPBCD.
After obtaining improved stability of Etanercept composition in phosphate -citrate buffer
other anti-aggregating agents from other class of compounds than amino acids were
tested.
It was observed that Urea and HPBCD also provided stable pharmaceutical compositions
of Etanercept in phosphate - citrate buffer. It is understood to mean that etanercept of the
pharmaceutical composition does not substantially lose its activity as compared to the
composition at the beginning of storage. The term 'substantially' refers to not more than
20%, or more preferably 15%, or even more preferably 10%, and most preferably 5% of
its activity relative to activity of the composition at the beginning of storage. The
pharmaceutical composition of the invention is suitable for long term storage. As used
herein, 'the long term storage' means that the storage of the pharmaceutical composition
is stable for more than a month, preferably more than 6 months or 12 months, more
preferably more than 24 months.
Tumor Necrosis Factor alpha (TNF-alpha) is a member of a group of cytokines that
stimulate the acute phase reaction, and thus is a cytokine involved in systemic
inflammation. TNF-alpha is able to induce inflammation, induce apoptotic cell death, and
to inhibit tumorgenesis and viral replication. Dysregulation of TNF-alpha production has
been implicated in a variety of human diseases like autoimmune disease, ankylosing
spondylitis, juvenile rheumatoid arthritis, psoriasis, psoriatic arthritis, rheumatoid
arthritis, Wegener's disease (granulomatosis), Crohn's disease or inflammatory bowel
disease, chronic obstructive pulmonary disease (COPD), Hepatitis C, endometriosis,
asthma, cachexia, atopic dermatitis, Alzheimer as well as cancer.
Dosage of the TNFR:Fc will depend on the disease, severity of condition, patient's
clinical history, and response to the (prior) therapy, and will be adjusted and monitored by
a physician. The pharmaceutical composition may be administered parenterally, such as
subcutaneously, intramuscularly, intravenously, intraperitoneally, intracerebrospinally,
intra-articularly, intrasynovially and/or intrathecally by either bolus injection or
continuous infusion.
In an embodiment the TNFR:Fc may be administered in adult or juvenile subject, wherein
the amount may range from about 1 - 80 mg. The dose may be administered once weekly,
twice weekly. Further, the doses may be administered weekly, biweekly, or separated by
several weeks e.g. three weeks. The therapeutic dose and duration may vary as per patient
response and patient requirement.
In another embodiment, a suitable regimen for juvenile and paediatric patients may
involve a dose of 0.4 mg/kg to 5 mg/kg of TNFR:Fc, administered one or more times per
week.
In case of adult rheumatoid arthritis, 25 mg twice weekly or 50 mg once weekly TNFR:Fc
may be administered.
In case of psoriatic arthiritis, 25 mg twice weekly or 50 mg once weekly TNFR:Fc may
be administered.
In case of Ankolysing spondylitis, 25 mg twice weekly or 50 mg once weekly TNFR:Fc
may be administered.
In case of adult plaque psoriasis, the recommended dose of TNFR:Fc is 25 mg
administered twice weekly or 50 mg administered once weekly. In case of pediatric
plaque psoriasis, the recommended dose of TNFR:Fc is 0.8 mg/Kg weekly with a
maximum of 50 mg dose per week.
In case of polyarticular juvenile idiopathic arthritis, the recommended dose of TNFR:Fc
is 0.8 mg/Kg weekly with a maximum of 50 mg dose per week.
In case of renal and hepatic impairment no dose adjustment is required.
In a second aspect, the invention relates to a kit comprising a composition according to
the first aspect and instructions for use of the present composition.
In a preferred embodiment, the composition is contained in a pre-filled syringe. In
another preferred embodiment, the composition is contained in a pre-filled vial. The kit
may comprise one or more unit dosage forms containing the pharmaceutical composition
of the invention.
Any suitable syringe or vial or cartridge may be used. The kit may also comprise the
pharmaceutical composition according to the invention in another secondary container,
such as in an autoinjector. The prefilled syringe may contain the composition in aqueous
form. Described syringe may be further supplied with an autoinjector, which often is a
disposable article for single use only, and may e.g. have a volume between 0.1 and 1 ml.
However, the syringe or autoinjector may also be for multi-usage or multi-dosing. The
described vial may contain the composition in lyophilised or aqueous state, and may serve
as a single or multiple use device. The vial may e.g. have a volume between 1 and 10 ml.
The pharmaceutical composition is sterile and stable for long period of time at 2-8 C.
Also it is stable upto 6 months when stored at 25°C. The invention provides
pharmaceutical composition essentially comprising of etanercept, phosphate - citrate
buffer, anti-aggregating agent selected from L-glycine, urea and HPBCD, a tonicity
modifier, a stabilizer and optionally other excipients in suitable combination thereof.
The invention further relates to a stable pharmaceutical composition, wherein the
composition is liquid or lyophilized. The invention is further related to a stable
pharmaceutical composition in a pre-filled syringe, vial, cartridge, or pen.
In an embodiment of the invention, the active pharmaceutical ingredient etanercept is
used which is obtained from recombinant DNA technology using CHO cells. The
concentration of the etanercept in the composition is 10 mg/mL to 100 mg/mL. In a
preferred embodiment of the invention, the concentration of etanercept in the composition
is 10 mg/mL to 60 mg/mL. In the most preferred embodiment of the invention, the
concentration of etanercept in the composition is 20 mg/mL to 60 mg/mL.
In another embodiment of the invention, the buffer is phosphate - citrate buffer. In an
embodiment of the invention, the concentration of the buffer in the composition is 10 mM
to 100 mM. In a preferred embodiment of the invention, the concentration of the buffer in
the composition is 10 mM to 50 mM. In another preferred embodiment of the invention,
the concentration of the buffer in the composition is 20 mM to 40 mM.
In another embodiment of the invention, the pH of the composition is 5 to 8.
In another embodiment of the invention, the etanercept composition comprises antiaggregating
agent selected from L-glycine, urea and HPBCD.
In an embodiment of the invention when the anti-aggregating agent is L-glycine, then the
concentration of L-glycine in the composition is 10 mM to 300 mM.
In another embodiment of the invention when the anti-aggregating agent is urea, the
concentration of urea in the composition is 20 mM to 50mM.
In another embodiment of the invention when the anti-aggregating agent is HPBCD, then
the concentration of HPBCD is 10 mM to lOOmM.
In another embodiment of the invention, the stable pharmaceutical composition further
comprises a parenterally acceptable tonicity agent. The tonicity agent is selected from the
group of salts such as sodium chloride, potassium chloride, calcium chloride or
saccharides such as mannitol, sucrose, glucose, or amino acids such as arginine, cysteine,
histidine and the like. The preferred tonicity agent is sodium chloride. The concentration
range varies from 0 mM to 150 mM.
In yet another embodiment of the invention, the stable pharmaceutical composition
further comprises a stabilizer. The stabilizer is selected from the group consisting of
sucrose, trehalose, lactose, mannitol.The preferred stabilizing agent is sucrose. The
concentration of the stabilizing agent in the composition varies from 0.5 wt to 10 wt .
In the most preferred embodiment of the invention, the concentration of the stabilizer in
the composition is 0.5 wt to 1.5 wt .
In yet another embodiment of the invention, the stable pharmaceutical composition may
optionally comprise a chelating agent. The chelating agent is selected from the group
consisting of EDTA, DTPA, HEDTA, NTA and TSP. The preferred chelating agent is
EDTA. In a more preferred embodiment of the invention, the concentration of EDTA is 0
mM to 10 mM.
In another embodiment of the invention, the stable pharmaceutical composition of the
invention comprises stable etanercept, phosphate - citrate buffer; anti-aggregating agent
selected from L-glycine, urea or HPBCD; sucrose as a stabilizing agent and with a long
shelf life at temperature 5°C .
In another embodiment of the invention, the stable pharmaceutical composition of the
invention comprises stable etanercept, phosphate - citrate buffer; anti-aggregating agent
selected from L-glycine, urea or HPBCD; sucrose as a stabilizing agent and with a long
shelf life at 5°C.
In another embodiment of the invention, the stable pharmaceutical composition of the
invention comprises stable etanercept, phosphate - citrate buffer; anti-aggregating agent
selected from L-glycine, urea or HPBCD; sucrose as a stabilizing agent and with 2 weeks
shelf life at 40°C.
In another embodiment of the invention, the stable pharmaceutical composition of the
invention comprises stable etanercept, phosphate - citrate buffer; anti-aggregating agent
selected from L-glycine, urea or HPBCD; sucrose as a stabilizing agent which provides
better stability to the pharmaceutical composition to maintain its activity for the longer
period of time providing longer shelf life.
In another embodiment, the invention pertains to a method of producing a pharmaceutical
composition according to the first aspect, comprising TNFR:Fc, phosphate - citrate
buffer, stabilizing agent selected from the group consisting of L-glycine, urea and
HPBCD.
In a preferred embodiment, the method may further comprise the step of adding at least
one tonicity modifier, such as sodium chloride; a stabilizer, such as sucrose and
optionally a chelating agent as defined above.
In another embodiment, the method may further comprise a lyophilization step, which
may be before or after adding the at least one tonicity modifier, and/or an excipient as
defined above.
Accordingly to a preferred embodiment of the present invention the pharmaceutical
composition comprises 10 mg/ml to 100 mg/ml of etanercept, about 10 mM to 100 mM of
phosphate citrate buffer, about 10 mM to 300 mM L-glycine, about 0 mM to 150 mM
sodium chloride and about 0.5 wt to 10wt sucrose having a pH range of 5 to 7.
Accordingly to another preferred embodiment of the present invention the pharmaceutical
composition comprises 10 mg/ml to 100 mg/ml of etanercept, about 10 mM to 100 mM of
phosphate citrate buffer, about 1 mg/ml to 18 mg/ml urea, about 1mM to 150 mM
sodium chloride, about 0.5 wt to 2 wt sucrose and about OmM to 10 mM
EDTAhaving a pH range of 5 to 7..
In another preferred embodiment of the present invention the pharmaceutical composition
comprises 10 mg/ml to 100 mg/ml of etanercept, about 10 mM to 100 mM of phosphate
citrate buffer, about 20 mg/ml to 30 mg/ml HPBCD, about 1mM to 150 mM sodium
chloride, about 0.5 wt to 2 wt sucrose and about 0 mM to 10 mM EDTAhaving a pH
range of 5 to 7..
The invention will be more fully understood by reference to the following examples.
However, the examples should not be construed as limiting the scope of the invention.
Experimental Section
The active ingredient etanercept, which was used for the described examples, is derived
from recombinant DNA technology in CHO cells. The CHO cells were cultured in a fedbatch
process. Etanercept was purified from the cell free harvest by standard purification
and filtration process including affinity chromatography and further chromatographic and
filtration steps. Etanercept was derived from different production batches which was used
in the examples, i.e., example 2 and example 5.
General Process for Preparation of stable pharmaceutical composition of Etanercept
The process for preparing theEtanercept drug substance compositions comprises of 2
steps viz. preparation of formulated bulk and fill finish. The formulated bulk is prepared
by diluting the drug substance with the formulation buffer to achieve the desired
concentration of drug product. The formulation buffer is prepared by adding required
quantity of Trisodium Citrate dihydrate and Sodium dihydrogen phosphate dihydrate to
WFI followed by mixing. Further, required quantities of other excipients are added to the
above solution and the desired volume is adjusted with WFI after adjustment of pH. The
formulation buffer is then aseptically filtered using 0.22 sterilizing grade PVDF filter.
As per the batch calculation, the required quantity of the Etanercept (in same formulation)
is aseptically diluted
The compositions were analysed by Size Exclusion - High-performance liquid
chromatography (SE-HPCL) at different time frames of storage at 5°C, 25°C and 40°C.
SE-HPLC separates the proteins and its related impurities on the basis of their size.
Therefore, it is useful to detect aggregation and fragmentation of Etanercept.
The examples which follow are illustrative of the invention and are not intended to be
limiting.
Example 1
The process for preparing theEtanercept drug substance compositions was comprises of 2
steps viz. preparation of formulated bulk and fill finish. The formulated bulk is prepared
by diluting the drug substance with the formulation buffer to achieve the desired
concentration of drug product. The formulation buffer is prepared by adding required
quantity (as mentioned in table 1) of Trisodium Citrate dihydrate and Sodium dihydrogen
phosphate dihydrate to WFI followed by mixing. Further, required quantities of Glycine
as anti-aggregating agent and other excipients are added to the above solution and the
desired volume is adjusted with WFI after adjustment of pH. The formulation buffer is
then aseptically filtered using 0.22 sterilizing grade PVDF filter. As per the batch
calculation, the required quantity of the Etanercept DS (in same formulation) is
aseptically diluted with the filtered formulation buffer to achieve the desired
concentration of 50 ± 5 mg/mL of Etanercept bulk. The formulated bulk is filtered
through 0.22 sterilizing grade PVDF filter and is aseptically dispensed into prefilled
syringes. The PFSs were then charged on stability at various temperatures.Table 1
describes the composition obtained using L-glycine as anti-aggregating agent in 75 mM
concentration.
Table 1: The composition of Example 1
Example 2 :
Etanercept compositions were studied in different buffers and using different antiaggregating
agents. The short term stability of Etanercept composition comprising
phosphate - citrate buffer with L-glycine as anti-aggregating agent was analysed at 5 °C
for 6 months and 40 °C for 2 weeks along with the Etanercept composition comprising
phosphate buffer with L-glycine as anti-aggregating agent, where other excipients were
maintained constant by using SE-HPLC and the results are provided in Table 2. The
Table 2 illustrates the stability studies of different etanercept compositions.
Table 2 : Stability studies at 5 °C and 40 °C
As table 2 illustrates, Etanercept composition comprising phosphate - citrate buffer with
L-glycine as anti-aggregating agent showed improved stability as compared to Etanercept
composition comprising phosphate buffer with L-glycine as anti-aggregating agent at 5
°C and 40 °C.
Example 3
The process for preparing the Etanercept drug substance composition is similar as
explained in example , wherein the anti-aggregating used is urea. Composition shown in
Table 3 was prepared using urea as anti-aggregating agent.
Table 3 : The composition of Example 3
Example 4
The process for preparing the Etanercept drug substance composition is similar as
explained in example , wherein the anti-aggregating used is HPBCD. Composition
shown in Table 4 was prepared using HPBCD as anti-aggregating agent.
Table 4 : The composition of Example 4
Example 5
The innovator composition of etanerceptcomprising phosphate buffer and arginine as
anti-aggregating agent (Composition 1) and the composition of etanercept comprising
phosphate - citrate buffer and L-glycine as anti-aggregating agent (Composition 2) were
filled in PFSs and were charged on long term stability which is ongoing. The data of
protein purity after 9 months storage at 5°C, 6 months storage at 25°C, 2 weeks storage at
40°C was analysed by using SE-HPCL and the results are provided in table 5.
Table 5 : Comparative stability data
Similarly, the compositions of example 3 and example 4, i.e., composition of etanercept
comprising phosphate - citrate buffer with urea as anti-aggregating agent and
composition of etanercept comprising phosphate - citrate buffer with HPBCD as antiaggregating
agent were filled in PFSs and were studied for stability at 5°, 25° and 40°C.
The stability of these compositions was assessed and was found comparable with the
composition 1 upto period of 2 weeks.
Example 6
Etanercept used for the lyophilization studies is formulated and dialyzed extensively with
pharmaceutical compositions mentioned in table 6. Respective formulated bulks are filled
in vials, half stoppered and are subjected to lyophilization.
Table 6 : The compositions of Example 6
All patents, patent applications and publications cited in this application are hereby
incorporated by reference in their entirety for all purposes to the same extent as if each
individual patent, patent application or publication were so individually denoted.
Although certain embodiments and examples have been described in detail above, those
having ordinary skill in the art will clearly understand that many modifications are
possible in the embodiments and examples without departing from the teachings thereof.
CLAIMS
I . A stable pharmaceutical composition comprising TNFR:Fc fusion protein and
phosphate - citrate buffer,.
2. A stable pharmaceutical composition comprising TNFR:Fc fusion protein, phosphate
- citrate buffer and anti-aggregating agent selected from the group consisting of Lglycine,
urea and 2-hydroxy propyl beta-cyclodextrin (HPBCD).
3. The composition as claimed in claim lor 2, wherein the pH of the composition is in
the range of 5 to 7.
4. The composition of any of the preceding claims wherein TNFR:Fc fusion protein is
Etanercept.
5. A stable pharmaceutical composition comprising TNFR:Fc fusion protein, phosphate
- citrate buffer, an anti-aggregating agent selected from L-glycine, urea and 2-
hydroxy propyl beta-cyclodextrin (HPBCD), a tonicity agent and a stabilizing agent.
6. The composition of claim 5, wherein the tonicity agent is selected from a group of
salts consisting of sodium chloride, potassium chloride, calcium chloride; group of
saccharides consisting mannitol, sucrose, glucose and amino acids.
7. The composition of claim 5, wherein the stabilizing agent is selected from the group
consisting of sucrose and trehalose.
8. The stable pharmaceutical composition of claim 5 comprising of etanercept,
phosphate-citrate buffer; L-glycine as anti-aggregating agent; sodium chloride as a
tonicity agent and sucrose as a stabilizing agent.
9. The stable pharmaceutical composition of claim 5 comprising etanercept, phosphatecitrate
buffer, urea as an anti-aggregating agent, sodium chloride as a tonicity agent,
sucrose as a stabilizing agent and EDTA as a chelating agent.
10. The stable pharmaceutical composition of claim 5 comprising etanercept, phosphatecitrate
buffer, HPBCD as an anti-aggregating agent, sodium chloride as a tonicity
agent, sucrose as a stabilizing agent and EDTA as a chelating agent.
I I . The composition of any of the preceding claims wherein the composition is sterile and
ready for parenteral administration.
12. The composition of claim 11, wherein the composition is liquid.
13. The composition of claim 11, wherein the composition is lyophilized.
14. The pharmaceutical composition of claim 8, comprising 10 mg/ml to 100 mg/ml of
etanercept, about 10 mM to 100 mM of phosphate citrate buffer, about 10 mM to 300
mM L-glycine, about 0 mM to 150 mM sodium chloride and about 0.5 wt to 10wt
sucrose.
15. The pharmaceutical composition of claim 9, comprising 10 mg/ml to 100 mg/ml of
etanercept, about 10 mM to 100 mM of phosphate citrate buffer, about 1 mg/ml to 18
mg/ml urea, about 1mM to 150 mM sodium chloride, about 0.5 wt to 2 wt
sucrose and about OmM to 10 mM EDTA.
16. The pharmaceutical composition of claim 10, comprising 10 mg/ml to 100 mg/ml of
etanercept, about 10 mM to 100 mM of phosphate citrate buffer, about 20 mg/ml to
30 mg/ml HPBCD, about 1mM to 150 mM sodium chloride, about 0.5 wt to 2 wt
sucrose and about 0 mM to 10 mM EDTA.
17. A kit comprising a composition of any of the preceding claims and instructions for
use of the said composition.
18. The Kit of claim 17, wherein the composition is liquid or lyophilized powder.
19. The kit of claim 17, wherein the composition is stored in a pre-filled sterile syringe or
vial or cartridge.
20. A method of treating a mammal in need there of comprising administering a
therapeutically effective amount of the pharmaceutical composition of any of claims 1
to 16.