The specific embodiment
Embodiment 1, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.4 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 6.8 phosphate buffer;
D.1.0mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of weight and number is 1: 1: 0.5.
Solution is behind 0.22 μ m membrane filtration, and is aseptic subpackaged.With aqueous injection place that humidity is 92%, temperature is incubator preserve separately 0 day, 5 days, 15 days, 20 days, January, February of 40 ℃, adopt then chicken dorsal ganglion method detect NGF biological activity, use pH meter to detect the injection pH value, detect aseptic situation and observe the transparency of solution according to method described in " Chinese biological goods rules 2000 editions ".As a result, the specific activity of the NGF of each time point be 4545,4545,3000,3000,1500,1500BU/ml (labelled amount: 3000BU/ml).The pH value of injection does not change during this period, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 2, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.10 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 6.5 phosphate buffer;
D.5.0mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of component is 1: 1: 1.
Injection storage condition and time are with embodiment 1.NGF is respectively 4545,4545,4545,3000,3000 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 3, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.15 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.10mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of weight and number is 1: 1: 1.
Injection storage condition and time are with embodiment 1.NGF is respectively 4545,4545,4545,4545,3000 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 4, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.20 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.4 phosphate buffer;
D.10mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of weight and number is 1: 1: 1.
Injection storage condition and time are with embodiment 1.NGF is respectively 4545,4545,4545,4545,3000 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 5, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.10mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and non-ionic surface active agent (tween 80), and the ratio of weight and number of three seed amino acids is 1: 1: 1; The concentration of tween 80 is 0.03%.
Injection storage condition and time are with embodiment 1.NGF is respectively 13636,4545,4545,4545,4545 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000 BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 6, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.20mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and non-ionic surface active agent (tween 80), and the ratio of weight and number of three seed amino acids is 1: 1: 1; The concentration of tween 80 is 0.01%.
Injection storage condition and time are with embodiment 1.NGF is respectively 13636,13636,4545,4545,4545 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 7, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.20mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine, histidine, lysine, NSC 334200 and non-ionic surface active agent (tween 80), and each amino acid whose parts by weight is identical; The concentration of tween 80 is 0.01%.
Injection storage condition and time are with embodiment 1.NGF is respectively 13636,13636,4545,4545,4545 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 8, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.4 phosphate buffer;
D.20mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and tween 80, and the ratio of weight and number of three seed amino acids is 1: 1: 1; The concentration of tween 80 is 0.005%.
Injection storage condition and time are with embodiment 1.NGF is respectively 13636,4545,4545,4545,4545 in the specific activity of each time point, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 9, remove albumin nerve growth factor aqueous injection
Pharmaceutical formulation:
A.100 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.4 phosphate buffer;
D.20mg/ml stabilizing agent;
E. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and tween 80, and the ratio of weight and number of three seed amino acids is 1: 1: 1; The concentration of tween 80 is 0.01%.
Injection storage condition and time are with embodiment 1.NGF is respectively 13636,13636,13636,4545,4545 in the specific activity of each time point, 3000BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative, and it is transparent that liquid keeps.
Embodiment 10, remove albumin nerve growth factor lyophilized injectable powder
A.4 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keeping the preparation pH value is 6.0 phosphate buffers;
D.1.0mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine, and three seed amino acid ratios of weight and number are 1: 1: 2.
Solution adopts the lyophilizing of frozen drying method behind 0.22 μ m membrane filtration.Earlier freeze dryer, freeze drying box temperature are reduced to-45 ℃, sample were put into pre-freeze 2 hours again, start vacuum pump, treat that vacuum reaches certain numerical value after, slowly open valve, close fridge, slowly be warming up to-25 ℃, treat water evaporates intact back intensification, in the time of 25 ℃, change drying again over to, capping then.
Injection storage condition and time are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 4545,4545,4545,4545,4545 in the specific activity of each time point NGF, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 11, remove albumin nerve growth factor lyophilized injectable powder
A.10 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 6.5 phosphate buffer;
D.5.0mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of weight and number is 1: 1: 1.
Injection storage condition and time and preparation technology are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 4545,4545,4545,4545,4545 in the specific activity of each time point NGF, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 12, remove albumin nerve growth factor lyophilized injectable powder
Pharmaceutical formulation:
A.15 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.10mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of weight and number is 1: 1: 1.
Injection storage condition and time and preparation technology are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 4545,4545,4545,4545,4545 in the specific activity of each time point NGF, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 13, remove albumin nerve growth factor lyophilized injectable powder
Pharmaceutical formulation:
A.20 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.4 phosphate buffer;
D.10mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine and glycine, and three's ratio of component is 1: 1: 1.
Injection storage condition and time are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 4545,4545,4545,4545,4545 in the specific activity of each time point NGF, 1500BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 14, remove albumin nerve growth factor lyophilized injectable powder
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.20mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and tween 80, and the ratio of component of three seed amino acids is 1: 1: 1; The concentration of tween 80 is 0.03%.
Injection storage condition and time are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 13636,13636,13636,4545,4545 in the specific activity of each time point NGF, 3000BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 15, remove albumin nerve growth factor lyophilized injectable powder
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.0 phosphate buffer;
D.20mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and tween 80 for stabilizing agent wherein, and the ratio of component of three seed amino acids is 1: 1: 2; The concentration of tween 80 is 0.01%.
Injection storage condition and time are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 13636,13636,4545,4545,4545 in the specific activity of each time point NGF, 3000BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 16, remove albumin nerve growth factor lyophilized injectable powder
Pharmaceutical formulation:
A.30 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.4 phosphate buffer;
D.20mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and tween 80, and the ratio of component of three seed amino acids is 1: 1: 2; The concentration of tween 80 is 0.005%.
Injection storage condition and time are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 13636,13636,4545,4545,4545 in the specific activity of each time point NGF, 3000BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 17, remove albumin nerve growth factor lyophilized injectable powder
Pharmaceutical formulation:
A.100 the nerve growth factor of μ g/ml;
B.8.5mg/ml sodium chloride;
C. keep the preparation pH value and be 7.4 phosphate buffer;
D.20mg/ml stabilizing agent;
E.60mg/ml mannitol;
F. the solution total amount adds to the water for injection of 0.66ml.
Wherein stabilizing agent is alanine, arginine, glycine and tween 80, and the ratio of component of three seed amino acids is 1: 1: 1; The concentration of non-ionic surface active agent is 0.01%.
Injection storage condition and time are with embodiment 1.Dissolve the powder pin in each time point with 0.66ml water for injection, and adopting embodiment 1 described method to detect the activity of NGF, the result is respectively 13636,13636,13636,4545,4545 in the specific activity of each time point NGF, 3000BU/ml (labelled amount: 3000BU/ml).PH value of injection does not change during this, and Bacteria Detection is negative.
Embodiment 18, embodiment 7 and embodiment 14 pharmaceutical formulations and existing albumin NGF prescription biological activity are relatively
By the foregoing description as can be known, the dosage performance of the stabilizing agent that embodiment 7 and embodiment 14 are selected is than best, so select for use itself and the existing albumin preparation that contains to make stability and compare (injection storage condition and time are with embodiment 1), result such as following table:
Table 1. embodiment 7,14 preparations contain albumin preparation stability relatively with existing
The prescription title | Different time points is surveyed slip-knot fruit (BU/ml) |
0 day | 5 days | 15 days | 20 days | January | February |
Embodiment 7 | ?13636 | ?4545 | ?4545 | ?4545 | ?4545 | ?3000 |
Embodiment 14 | ?13636 | ?13636 | ?4545 | ?4545 | ?4545 | ?3000 |
Existing preparation | 13636 | ?13636 | ?4545 | ?4545 | ?4545 | ?3000 |
The present invention selects for use aminoacid or aminoacid and non-ionic surface active agent as stabilizing agent, make the NGF preparation can not only make lyophilized preparation and can also be prepared into water preparation easy to use, the blood product pollution of effectively having avoided the human serum albumin to cause, solve the quantitative difficult problem of NGF, and reduced production cost; Experiment showed, that by comparing the stabilizing agent that the present invention selects for use can reach stablizing effect same as the prior art, can replace the human serum albumin fully with existing product.