CA2215936A1 - Hcg liquid formulations - Google Patents
Hcg liquid formulations Download PDFInfo
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- CA2215936A1 CA2215936A1 CA002215936A CA2215936A CA2215936A1 CA 2215936 A1 CA2215936 A1 CA 2215936A1 CA 002215936 A CA002215936 A CA 002215936A CA 2215936 A CA2215936 A CA 2215936A CA 2215936 A1 CA2215936 A1 CA 2215936A1
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- hcg
- liquid pharmaceutical
- pharmaceutical composition
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- mannitol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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Abstract
The invention refers to liquid pharmaceutical compositions containing hCG stabilised with a polyalcohol or a non-reducing sugar. Preferably, the compositions are stabilised with mannitol. In the preferred embodiments such compositions are aqueous solutions in a phosphate buffer at pH 7. Such compositions are ready to be injected and, therefore, the step of reconstitution of the lyophilised powder is avoided, thus simplifying the way of use.
Description
CA 0221~936 1997-09-19 WO 96/29095 PCT/~P95/01055 HCG LIQUID FORMULATIONS
The present invention relates to gonadotropin co~.L~ P liquid ph~rm~ceutical compositions. More precisely, it concerns liquid 5 form~ tions of hCG (human Chorionic Gonadotropin) stablised with a polyalcohol or a non-reducing sugar.
It is known that highly purified proteins easily undergo degradation, even due to the contact with atmospheric agents. This characteristic is even more evident for proteins produced by recombinant DNA techniques.
o Such proteins are usually stabilised with saccharides, such as lactose, or with mannitol, or else with proteins or aminoacids, such as albumin and glycin.
The injectable stabilised formulations of gonadotropins are obtained with a process which includes always a step of lyophili.~on to obtain a dry 15 powder; in such a way the stabilised formulations are able to m~int~in a longer cycle life, even if stored at room temperature.
WO 93/11788 describes lyophilised gonadotropin-co--l~N~ g ph~ ceutical compositions stabilised with sucrose, alone or in combination with other stabilising agents. In said patent application it is 20 shown that the stability provided to the lyophilised compositions under study by sucrose was better than that provided by lactose or mannitol.
No liquid stabilised formulations of gonadotropins have been described until now. It is highly desirable to obtain such liquid formulations so as to have the compositions ready to be injected and to avoid the 25 reconstitution of the lyophilised powder, thus simplifying the way of use.
We have surprisingly found that it is possible to obtain such liquid stabilised form~ hons.
The main object of the present invention is to provide a liquid phzlrm~ceutical composition co--~ -P hCG stabilised with a polyalcohol 30 or a non-reducing sugar. Preferably the polyalcohol is mannitol and the non-retlll~inP sugar is sucrose. More ~ler~l~bly the liquid formulations of the invention are stabilised with m~nnitol.
The solution is pr~r~lably a buffered aqueous solution and the buffer accordin to the invention is selected from the group consisting of CONFIRMATION COPY
CA 0221~936 1997-09-19 phosphate, acetate or succinate buffer. The ~lt;r~lled buffer is phosphate and the pH is ~lcr~l~bly 7.00.
The hCG is ~l~r~l~bly recomhin~nt and can be prepared, for ~-x~mple, by expression in CHO (Chinese ~m.~t~r Ovary) cells, transformed 5 with the corresponding DNA, according to the technique described in ~ European Patent 160699.
A further object of the present invention is to provide a process for the preparation of said liquid ph~rm~ceutical composition comprising diluting a hCG bulk solution in a buffer solution co~ g the excipients.
Still another object of the present invention is to provide a form of presentation of said liquid ph~Tm~ceu~cal composition comprising such foTm~ ~ion hermetically closed in a sterile condition in a container suitable for the storage before the use.
In order to optimise the stability of the hCG formulations of the invention a series of prelimin~ry experiments have been carried out with different buffers at various pH, ionic strength, dielectric constant and concentration of rec-hCG.
In order to evaluate the effect of pH and of the buffer, 0.01 M
solutions of phosphate, succinate or acetate buffers were prepared with water for injection. The pH was adjusted to 6.0, 7.0 or 8.0 with NaOH l M.
The bulk solution of rec-hCG was added to the buffer systems to obtain solutions at 5,000 IU/ml. The solutions were then filtered and poured into 3 ml glass vials. The composition of the formulations thus prepared is reported in Table 1. The accelerated stability of these formlll~hons has been studied, So that the stability of the same can be foreseen when they are stored in containers at room temperature, through the extrapolation of the data at higher temperatures. In this case the samples were stored at 40~ and 50~C
and the stability of rec-hCG was checked by d~ g its purity by HPSEC analyses according to the following standard conditions:
30 Phase A 0.1 M phosphate pH 6.7 + 0.1 M Na2S~4 Isocratic conditions 100% phase A
Column TSK G 2000 SVVXL
FlowRate 0.5 ml/min W Detector 214 nm 35 Injection Volume 20 ~1(10,000 IU strength) 40 ,~ (5,000 IU strength) Table 3 l~oll~ the perc~Mt~e of rec-hCG monomer peak delelll.ined by HPSEC. The results show that the solutions at pH 6.0 and 8.0 are less stable in comparison with the solutions at pH 7.0 and that no rem~rk~ble stability differences were observed among the burr~
The effect of the ionic strength was evaluated on rec-hCG 5,000 IU/ml solutions, prepared with phosphate and succinate buffers 0.01M at pH
7.0, adjusted with NaCl to the following values of osmolality: 150, 300 and 400 mOsm. The composition of the form~ tions is reported in Table 2. The o samples were stored at 4~, 25~, 40~ and 50~C and tested for the stability ofrec-hCG by HPSEC. The results, reported in table 4, show that the increase of ionic strength negatively affects the stability of rec-hCG.
The effect of the dielectric constant was evaluated on 5,000 IU/ml solutions of rec-hCG, prepared with phosphate and succinate buffers 0.01 M
at pH 7, cont~inin~ 5, 10 and 15% propylene glycol. The composition of the formulations is reported on Table 2. The samples were stored at 4~, 25~, 40~ and 50~C and tested for the stability of rec-hCG by HPSEC. The results, reported in Table 4, show that increasing the percentage of propylene glycol negatively affects the stability of rec-hCG.
In order to evaluate the effect of the rec-hCG concentration, the stability at 50~C of the solutions in phosphate buffer 0.01 M at pH 7.0, cont~inin~ respectively 2,500, 5,000, 7,500 and 10,000 IU/ml of rec-hCG
was monitored by HPSEC for 2 weeks. The results reported in Table 5 showed that the stability was higher for the more concentrated solutions In order to compare the effects of various stabilisers and/or excipients on the stability of rec-hCG, six liquid formulations, in phosphate buffer 0. 01 M at pH 7.0 cont~;nin~ 10,000 IU/ml rec-hCG were l~lc~alcd, as a first step. Sucrose, glycine, glucose, mannitol, lactose and NaCl were used, as stabilisers/excipient. The composition of the formulations is reported in Table 6. These formulations were submitted to the stability tests by storing samples at 4~, 25~, 40~ and 50~C and tested by a Bioassay and HPSEC.
Subsequently, based on the results of said first step, four lots of two selectedliquid formulations were prepared, using as stabilisers sucrose and mannitol.
Table 7 reports the composition of such formulations.
The Bioassay has been carried out in accordance with the European Ph~rm~copoeia Monograph.
In Table 8 the HPSEC stability data are reported and in Table 9 the values of bioactivity are reported . The results showed the following:
5 1. the bioactivity of the form~ hons cont~ining glucose and lactose rem~rk~bly decreased at 50~C after 1 week storage. Also monalmer peak was lower colllp~u~d to that measured in the other form~ tions.
The present invention relates to gonadotropin co~.L~ P liquid ph~rm~ceutical compositions. More precisely, it concerns liquid 5 form~ tions of hCG (human Chorionic Gonadotropin) stablised with a polyalcohol or a non-reducing sugar.
It is known that highly purified proteins easily undergo degradation, even due to the contact with atmospheric agents. This characteristic is even more evident for proteins produced by recombinant DNA techniques.
o Such proteins are usually stabilised with saccharides, such as lactose, or with mannitol, or else with proteins or aminoacids, such as albumin and glycin.
The injectable stabilised formulations of gonadotropins are obtained with a process which includes always a step of lyophili.~on to obtain a dry 15 powder; in such a way the stabilised formulations are able to m~int~in a longer cycle life, even if stored at room temperature.
WO 93/11788 describes lyophilised gonadotropin-co--l~N~ g ph~ ceutical compositions stabilised with sucrose, alone or in combination with other stabilising agents. In said patent application it is 20 shown that the stability provided to the lyophilised compositions under study by sucrose was better than that provided by lactose or mannitol.
No liquid stabilised formulations of gonadotropins have been described until now. It is highly desirable to obtain such liquid formulations so as to have the compositions ready to be injected and to avoid the 25 reconstitution of the lyophilised powder, thus simplifying the way of use.
We have surprisingly found that it is possible to obtain such liquid stabilised form~ hons.
The main object of the present invention is to provide a liquid phzlrm~ceutical composition co--~ -P hCG stabilised with a polyalcohol 30 or a non-reducing sugar. Preferably the polyalcohol is mannitol and the non-retlll~inP sugar is sucrose. More ~ler~l~bly the liquid formulations of the invention are stabilised with m~nnitol.
The solution is pr~r~lably a buffered aqueous solution and the buffer accordin to the invention is selected from the group consisting of CONFIRMATION COPY
CA 0221~936 1997-09-19 phosphate, acetate or succinate buffer. The ~lt;r~lled buffer is phosphate and the pH is ~lcr~l~bly 7.00.
The hCG is ~l~r~l~bly recomhin~nt and can be prepared, for ~-x~mple, by expression in CHO (Chinese ~m.~t~r Ovary) cells, transformed 5 with the corresponding DNA, according to the technique described in ~ European Patent 160699.
A further object of the present invention is to provide a process for the preparation of said liquid ph~rm~ceutical composition comprising diluting a hCG bulk solution in a buffer solution co~ g the excipients.
Still another object of the present invention is to provide a form of presentation of said liquid ph~Tm~ceu~cal composition comprising such foTm~ ~ion hermetically closed in a sterile condition in a container suitable for the storage before the use.
In order to optimise the stability of the hCG formulations of the invention a series of prelimin~ry experiments have been carried out with different buffers at various pH, ionic strength, dielectric constant and concentration of rec-hCG.
In order to evaluate the effect of pH and of the buffer, 0.01 M
solutions of phosphate, succinate or acetate buffers were prepared with water for injection. The pH was adjusted to 6.0, 7.0 or 8.0 with NaOH l M.
The bulk solution of rec-hCG was added to the buffer systems to obtain solutions at 5,000 IU/ml. The solutions were then filtered and poured into 3 ml glass vials. The composition of the formulations thus prepared is reported in Table 1. The accelerated stability of these formlll~hons has been studied, So that the stability of the same can be foreseen when they are stored in containers at room temperature, through the extrapolation of the data at higher temperatures. In this case the samples were stored at 40~ and 50~C
and the stability of rec-hCG was checked by d~ g its purity by HPSEC analyses according to the following standard conditions:
30 Phase A 0.1 M phosphate pH 6.7 + 0.1 M Na2S~4 Isocratic conditions 100% phase A
Column TSK G 2000 SVVXL
FlowRate 0.5 ml/min W Detector 214 nm 35 Injection Volume 20 ~1(10,000 IU strength) 40 ,~ (5,000 IU strength) Table 3 l~oll~ the perc~Mt~e of rec-hCG monomer peak delelll.ined by HPSEC. The results show that the solutions at pH 6.0 and 8.0 are less stable in comparison with the solutions at pH 7.0 and that no rem~rk~ble stability differences were observed among the burr~
The effect of the ionic strength was evaluated on rec-hCG 5,000 IU/ml solutions, prepared with phosphate and succinate buffers 0.01M at pH
7.0, adjusted with NaCl to the following values of osmolality: 150, 300 and 400 mOsm. The composition of the form~ tions is reported in Table 2. The o samples were stored at 4~, 25~, 40~ and 50~C and tested for the stability ofrec-hCG by HPSEC. The results, reported in table 4, show that the increase of ionic strength negatively affects the stability of rec-hCG.
The effect of the dielectric constant was evaluated on 5,000 IU/ml solutions of rec-hCG, prepared with phosphate and succinate buffers 0.01 M
at pH 7, cont~inin~ 5, 10 and 15% propylene glycol. The composition of the formulations is reported on Table 2. The samples were stored at 4~, 25~, 40~ and 50~C and tested for the stability of rec-hCG by HPSEC. The results, reported in Table 4, show that increasing the percentage of propylene glycol negatively affects the stability of rec-hCG.
In order to evaluate the effect of the rec-hCG concentration, the stability at 50~C of the solutions in phosphate buffer 0.01 M at pH 7.0, cont~inin~ respectively 2,500, 5,000, 7,500 and 10,000 IU/ml of rec-hCG
was monitored by HPSEC for 2 weeks. The results reported in Table 5 showed that the stability was higher for the more concentrated solutions In order to compare the effects of various stabilisers and/or excipients on the stability of rec-hCG, six liquid formulations, in phosphate buffer 0. 01 M at pH 7.0 cont~;nin~ 10,000 IU/ml rec-hCG were l~lc~alcd, as a first step. Sucrose, glycine, glucose, mannitol, lactose and NaCl were used, as stabilisers/excipient. The composition of the formulations is reported in Table 6. These formulations were submitted to the stability tests by storing samples at 4~, 25~, 40~ and 50~C and tested by a Bioassay and HPSEC.
Subsequently, based on the results of said first step, four lots of two selectedliquid formulations were prepared, using as stabilisers sucrose and mannitol.
Table 7 reports the composition of such formulations.
The Bioassay has been carried out in accordance with the European Ph~rm~copoeia Monograph.
In Table 8 the HPSEC stability data are reported and in Table 9 the values of bioactivity are reported . The results showed the following:
5 1. the bioactivity of the form~ hons cont~ining glucose and lactose rem~rk~bly decreased at 50~C after 1 week storage. Also monalmer peak was lower colllp~u~d to that measured in the other form~ tions.
2. in the presence of glycine and NaCl a more evident decrease of bioactivity and of purity was measured in comparison to the formulations CO~I~;.i.,i.. g sucrose and mannitol. Also in this case the decrease of the percentage of the rec-hCG monomer peak, was not due to the iFormation of aggregates forms, but to the increase of free subunits.
Tables 10 and 11 report the purity d~Lt;lmilled by HPSEC for the 5,000 and 10,000 IU strength respectively. Data show that even after three 15 weeks at 50~C the purity is higher in the formulations con~ lg mannitol compared to the formulations Cont~inin,o sucrose. Tables 12 and 13 report the purity of the a subunit detell~ ed by reverse phase HPLC after 1 week storage at 50~C for the sucrose and mannitol formulations. The data confirm the better stability of the formulation cont~ining mannitol in comparison to 20 that co.,L~ g sucrose.
Reverse Phase HPLC analyses have been performed ~,vith the following standard conditions:
Phase A 1 ml TFA in 1 liter of bidistilled water Phase B 0. 79 ml TFA in 1 liter of acetonitrile 25 Gradient conditions time A% B%
20' 60 40 21' 20 80 22' 85 15 30 Column Aquapore RP 300 25 cm Column temperature 40~C
Flow Rate 1 ml/min W detector 214 nm l~jection volume 10 ,ul _ CA 0221~936 1997-09-19 ~ the Tables 14 and 15 the results of the bioactivity assay are reported. No appreciable bioactivity decrease was observed after 24 weeks at 4~ and 25~C in the mannitol form~ on.
According to the present invention, the liquid ph~rm~ce~ cal compositions contain from 1,000 to 40,000 lU/ml, l,lert;ldbly 10,000 IU/ml, of h~G and from 10 to 180 mg/l, ~;re~dbly 54.6 mg/l, of mannitol in a 0.01 M buffer solution.
EXAMPLES OF PHA~MACEUTICAL MANIJFACTURING
M~qt~-.ri~ Phosphoric acid 85% RPE ACS (Carlo Erba); Mannitol DAB, Ph Eur BP, FU, USP, FCC, E421 (Merck), NaOH 1 M (Merck), water for injections.
The primary container for the formulated vials consists of:
Tables 10 and 11 report the purity d~Lt;lmilled by HPSEC for the 5,000 and 10,000 IU strength respectively. Data show that even after three 15 weeks at 50~C the purity is higher in the formulations con~ lg mannitol compared to the formulations Cont~inin,o sucrose. Tables 12 and 13 report the purity of the a subunit detell~ ed by reverse phase HPLC after 1 week storage at 50~C for the sucrose and mannitol formulations. The data confirm the better stability of the formulation cont~ining mannitol in comparison to 20 that co.,L~ g sucrose.
Reverse Phase HPLC analyses have been performed ~,vith the following standard conditions:
Phase A 1 ml TFA in 1 liter of bidistilled water Phase B 0. 79 ml TFA in 1 liter of acetonitrile 25 Gradient conditions time A% B%
20' 60 40 21' 20 80 22' 85 15 30 Column Aquapore RP 300 25 cm Column temperature 40~C
Flow Rate 1 ml/min W detector 214 nm l~jection volume 10 ,ul _ CA 0221~936 1997-09-19 ~ the Tables 14 and 15 the results of the bioactivity assay are reported. No appreciable bioactivity decrease was observed after 24 weeks at 4~ and 25~C in the mannitol form~ on.
According to the present invention, the liquid ph~rm~ce~ cal compositions contain from 1,000 to 40,000 lU/ml, l,lert;ldbly 10,000 IU/ml, of h~G and from 10 to 180 mg/l, ~;re~dbly 54.6 mg/l, of mannitol in a 0.01 M buffer solution.
EXAMPLES OF PHA~MACEUTICAL MANIJFACTURING
M~qt~-.ri~ Phosphoric acid 85% RPE ACS (Carlo Erba); Mannitol DAB, Ph Eur BP, FU, USP, FCC, E421 (Merck), NaOH 1 M (Merck), water for injections.
The primary container for the formulated vials consists of:
3 ml glass vials (DIN 2R) (borosilicate glass type I), Rubber closures (Ph~rm~gllmmi W1816 V50), Aluminium rings and flip off caps (Pharma Metal GmbH).
Preparation of rec-hCG solution co~ mannitol The phosphoric acid (0.98 g) is added to the water for injections (600 ml). If necessary, the pH is adjusted to 7.0 with NaOH 1 M. Mannitol (54.6 g) is added to the phosphoric acid solution and the pH is again checked and, if necessary, adjusted to 7.00 + 0.2 with NaOH 1 M or with phosphoric acid diluted 1:5. The rec-hCG bulk (l0 MIU or 20 MIU, if the final desired strenght is 5,000 or 10,000 IU respectively) is then added to the excipient solution and the pH is again checked and, if necessary, adjusted to 7.00 + 0.2 with NaOH 1 M or with phosphoric acid diluted 1:5.
The solution is brought to 1 liter with water for injections. Such solution is then filtered through a 0.22 ~m Millipak 20 filter under a pressure not higher than 1.5 atm, under l~min~r flow, collecting the solution intoa flask and stirring gently for about 1 minute.
The vials are then filled up with 0.5 ml of the rec-hCG solution.
CA 02215936 1997-09-l9 TABLE 1 - COMPOSlTION OF r-hCG SOLUTIONS
p~:/buffer effect Acetate buffer solution Amountlml r-hCG bulk 5000 IU
Acetic acid ~lacial 0.6 m g NaOE lM q.s. to p H 6.0,7.0,8.0 Succinate buffer solution r-hCG bulk 5000 IU
Succinic acid 1.18 m g NaOEI lM q.s. to p H 6.0,7.0,8.0 Phosphate buffer solution r-hCG bu~ 5000 IU
Phosphoric acid 85% 0.98 m g NaOE lM q.s. to p H 6.0,7.0,8.0 F~ling volu m e: 1 n ~
WO 96/29095 PCTIEP9S/OlOSS
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' 3"'' ' ~3 --O O O O O ~,o ~-~ 11 11 11 4 4 4 ~ V o -5 PCT~P95/01055 TABLE 3 - r-hCG PURlTY (%) ElPSEC DATA
'VB'_, t~ ' e -~c . . . . . 1 _ ,:
,.................. :
ACE/6 100 95.85 92.70 84.99 97.51 94.10 ACE/7 100 96.62 93.26 88.02 97.27 94.05 ACE/8 100 96.51 92.70 87.10 97.45 95.12 SUC/6 100 94.56 91.28 82.11 96.92 93. I l SUC/7 100 95.78 94.20 88.05 96.91 93.99 SUC/8 100 95.36 90.12 83.00 97.61 94.02 FOS/6 100 94.10 90.76 81.00 97.50 93.00 FOS/7 100 96.09 93.12 86.93 96.72 93.74 FOS/8 100 94.21 82.52 74.96 96.77 93.55 W = week ACE = acetate buffer SUC = succ~nate buffer FOS = phosphate buffer 6/7/8 = pH 6 0, 7.0, 8.0 =
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CA 02215936 1997-09-l9 ~, .~", r~ ~
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TABLE 5 - r-hCG PURITY (%) ~IPSEC DATA
cn-cer~rh~ e ec~
.. . . ~ -~, , .. , ,, ., , :
Fos /2500 100 87.3 84.0 Fos/5000 1 00 90. 8 89.1 Fos/7500 100 92.9 89.8 Fos/10000 100 92.5 90.9 Fos/2500: 2,500 IU/ ml of r-hCG
Fos/5000 :5,000 IU/ ml of r-hCG
Fos/7500: 7,500 IU/ ml of r-hCG
Fos/10000:10,000 IU/ml of r-hCG ~
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TABLE~ 7 - LIQUID FORMllLATIONS
Vial compositioll r-hCG IU/m 10,000 20,000 SUCROS~ mg/ml 102.6 102.6 O. PHOSPHORIC ACID mglml 0.98 0.98 SODIUM q.s. to pH 7.0q.s. to pH 7.0 HYDROXIDE
,~::::
r-hCG IU/ml 10,000 20,000 MANNITOL mg/ml 54.6 54.6 O. PHOSPEIORIC ACID mg/ml 0.98 0.98 SOD~UM q.s. to pH 7.0q.s. to pH 7.0 HYDROXIDE
Filling volume: 0. 5 ml -~ ~ E- ~ E~ o o o Z; o Z o ~ o Z o Z ~o ~'' O O E~ O E-- ~
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'' ~' o o ~ o~ z o _ % Z Z ~' ,,.',', ~, O~ G' Z ~ Z ~ ;~
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~ ~ ~
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.... , ......... ... : = . ~ . ,.~ . .
:: . : .. , .. . . , . ,, , .. .. .: :
H C G/5000/S01 100 90.0 86.3 97.2 H C G/5000/ M 01 100 89.5 88.3 97.6 W =week SO l=sucrose M0 1--~ol TABLE 11- LIQUID FORMULATIO~: CONC. 10,000 IU/vial EP-SI~C Stability data: purity (%) Formulation development . .. .... ... .. ........ . . . . . ...... .. .
,.,,'.. '.'..... ;'.-.', .. .. .
H C G/10000/S01 100 91.8 88.9 97.9 H C G/10000/ M 01 100 93.4 92.1 97.2 0 W = week SO l=sucrose M0 1-- ~a~ .lol CA 022l5936 l997-09-l9 a subunit purity by RP-~LC
EICG/5000/S01 100 90.2 a(%) l~CG/5000/M01 100 94.7 a(%) W= week S01 = Sucrose M01--Mannitol TABLI~ 13 - LIQUII) FORMUI~TION
a subunit purity by RP-HPLC
,, .,, ".: ,.,,,: . . ,~ , . :
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ECG/10000/S01 100 92.4 a(~/O) ~CG/10000/M01 l00 95. l a(%) W= week S0l = Sucrose M0l= MaIInitol . ..
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Preparation of rec-hCG solution co~ mannitol The phosphoric acid (0.98 g) is added to the water for injections (600 ml). If necessary, the pH is adjusted to 7.0 with NaOH 1 M. Mannitol (54.6 g) is added to the phosphoric acid solution and the pH is again checked and, if necessary, adjusted to 7.00 + 0.2 with NaOH 1 M or with phosphoric acid diluted 1:5. The rec-hCG bulk (l0 MIU or 20 MIU, if the final desired strenght is 5,000 or 10,000 IU respectively) is then added to the excipient solution and the pH is again checked and, if necessary, adjusted to 7.00 + 0.2 with NaOH 1 M or with phosphoric acid diluted 1:5.
The solution is brought to 1 liter with water for injections. Such solution is then filtered through a 0.22 ~m Millipak 20 filter under a pressure not higher than 1.5 atm, under l~min~r flow, collecting the solution intoa flask and stirring gently for about 1 minute.
The vials are then filled up with 0.5 ml of the rec-hCG solution.
CA 02215936 1997-09-l9 TABLE 1 - COMPOSlTION OF r-hCG SOLUTIONS
p~:/buffer effect Acetate buffer solution Amountlml r-hCG bulk 5000 IU
Acetic acid ~lacial 0.6 m g NaOE lM q.s. to p H 6.0,7.0,8.0 Succinate buffer solution r-hCG bulk 5000 IU
Succinic acid 1.18 m g NaOEI lM q.s. to p H 6.0,7.0,8.0 Phosphate buffer solution r-hCG bu~ 5000 IU
Phosphoric acid 85% 0.98 m g NaOE lM q.s. to p H 6.0,7.0,8.0 F~ling volu m e: 1 n ~
WO 96/29095 PCTIEP9S/OlOSS
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'VB'_, t~ ' e -~c . . . . . 1 _ ,:
,.................. :
ACE/6 100 95.85 92.70 84.99 97.51 94.10 ACE/7 100 96.62 93.26 88.02 97.27 94.05 ACE/8 100 96.51 92.70 87.10 97.45 95.12 SUC/6 100 94.56 91.28 82.11 96.92 93. I l SUC/7 100 95.78 94.20 88.05 96.91 93.99 SUC/8 100 95.36 90.12 83.00 97.61 94.02 FOS/6 100 94.10 90.76 81.00 97.50 93.00 FOS/7 100 96.09 93.12 86.93 96.72 93.74 FOS/8 100 94.21 82.52 74.96 96.77 93.55 W = week ACE = acetate buffer SUC = succ~nate buffer FOS = phosphate buffer 6/7/8 = pH 6 0, 7.0, 8.0 =
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TABLE 5 - r-hCG PURITY (%) ~IPSEC DATA
cn-cer~rh~ e ec~
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Fos /2500 100 87.3 84.0 Fos/5000 1 00 90. 8 89.1 Fos/7500 100 92.9 89.8 Fos/10000 100 92.5 90.9 Fos/2500: 2,500 IU/ ml of r-hCG
Fos/5000 :5,000 IU/ ml of r-hCG
Fos/7500: 7,500 IU/ ml of r-hCG
Fos/10000:10,000 IU/ml of r-hCG ~
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TABLE~ 7 - LIQUID FORMllLATIONS
Vial compositioll r-hCG IU/m 10,000 20,000 SUCROS~ mg/ml 102.6 102.6 O. PHOSPHORIC ACID mglml 0.98 0.98 SODIUM q.s. to pH 7.0q.s. to pH 7.0 HYDROXIDE
,~::::
r-hCG IU/ml 10,000 20,000 MANNITOL mg/ml 54.6 54.6 O. PHOSPEIORIC ACID mg/ml 0.98 0.98 SOD~UM q.s. to pH 7.0q.s. to pH 7.0 HYDROXIDE
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.... , ......... ... : = . ~ . ,.~ . .
:: . : .. , .. . . , . ,, , .. .. .: :
H C G/5000/S01 100 90.0 86.3 97.2 H C G/5000/ M 01 100 89.5 88.3 97.6 W =week SO l=sucrose M0 1--~ol TABLE 11- LIQUID FORMULATIO~: CONC. 10,000 IU/vial EP-SI~C Stability data: purity (%) Formulation development . .. .... ... .. ........ . . . . . ...... .. .
,.,,'.. '.'..... ;'.-.', .. .. .
H C G/10000/S01 100 91.8 88.9 97.9 H C G/10000/ M 01 100 93.4 92.1 97.2 0 W = week SO l=sucrose M0 1-- ~a~ .lol CA 022l5936 l997-09-l9 a subunit purity by RP-~LC
EICG/5000/S01 100 90.2 a(%) l~CG/5000/M01 100 94.7 a(%) W= week S01 = Sucrose M01--Mannitol TABLI~ 13 - LIQUII) FORMUI~TION
a subunit purity by RP-HPLC
,, .,, ".: ,.,,,: . . ,~ , . :
,,.','''',''.. ,','.',, '' ' ' i ~
ECG/10000/S01 100 92.4 a(~/O) ~CG/10000/M01 l00 95. l a(%) W= week S0l = Sucrose M0l= MaIInitol . ..
. _ * --~
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Claims (9)
1. A stable, liquid pharmaceutical composition comprising recombinant human Chorionic Gonadotropin and a stabilizing amount of mannitol.
2. A liquid pharmaceutical composition according to Claim 1, wherein the solution is a buffered aqueous solution.
3. A liquid pharmaceutical composition according to Claim 2, wherein the buffer solution is selected from the group consisting of acetate, succinate and phosphate buffer.
4. A liquid pharmaceutical composition according to Claim 3, wherein the buffer is phosphate buffer.
5. A liquid pharmaceutical composition according to any of Claims 2 to 4, wherein the buffer solution is at pH 7.00.
6. A liquid pharmaceutical composition according to any of Claims 2 to 5, wherein the buffer solution is 0.01 M
7. A liquid pharmaceutical composition according to Claim 1, comprising from 1,000 to 40,000 IU/ml of hCG and from 10 to 180 mg/l of mannitol in a 0.01 M phosphate buffer at pH 7.00.
8. A process for the preparation of a liquid pharmaceutical composition according to Claim 1, comprising diluting a hCG bulk solution in a buffer solution containing the excipients.
9. A form of presentation of a liquid pharmaceutical composition of Claim 1 hermetically closed in a sterile condition in a container suitable for the storage before the use.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002215936A CA2215936A1 (en) | 1995-03-21 | 1995-03-21 | Hcg liquid formulations |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002215936A CA2215936A1 (en) | 1995-03-21 | 1995-03-21 | Hcg liquid formulations |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2215936A1 true CA2215936A1 (en) | 1996-09-26 |
Family
ID=4161498
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002215936A Abandoned CA2215936A1 (en) | 1995-03-21 | 1995-03-21 | Hcg liquid formulations |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2215936A1 (en) |
-
1995
- 1995-03-21 CA CA002215936A patent/CA2215936A1/en not_active Abandoned
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Discontinued |