CN108159404B - Recombinant human parathyroid hormone preparation and preparation method thereof - Google Patents
Recombinant human parathyroid hormone preparation and preparation method thereof Download PDFInfo
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- CN108159404B CN108159404B CN201810013546.9A CN201810013546A CN108159404B CN 108159404 B CN108159404 B CN 108159404B CN 201810013546 A CN201810013546 A CN 201810013546A CN 108159404 B CN108159404 B CN 108159404B
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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
- A61K38/29—Parathyroid hormone (parathormone); Parathyroid hormone-related peptides
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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
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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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
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
Abstract
The present invention relates to field of pharmaceutical preparations, a kind of recombinant human parathyroid hormone preparation and preparation method thereof is provided.The recombinant human parathyroid hormone preparation, it is the preparation liquid for mixing raw material and water for injection, raw material includes the buffer for being 8-30mmol/L in the concentration prepared in liquid in the recombinant human parathyroid hormone stoste that the concentration prepared in liquid is 10-500 μ g/ml, the osmotic pressure regulator that the concentration in liquid is 40-50mg/ml is being prepared, the bacteriostatic agent that the concentration in liquid is 2.4-3.3mg/ml is being prepared.The preparation method includes being uniformly mixed to form preparation liquid with water for injection by above-mentioned raw materials.Recombinant human parathyroid hormone preparation provided in this embodiment is with good stability at room temperature, has better safety.In addition, the preparation method is simple, operation is easy, easy to accomplish.
Description
Technical field
The present invention relates to field of pharmaceutical preparations, in particular to a kind of recombinant human parathyroid hormone preparation and its system
Preparation Method.
Background technique
With the extension of human longevity, aging of population more becomes obviously, and osteoporosis will become large-scale global strong
Kang Wenti.According to European Union 1998, osteoporotic fracture lifetime risk, women was up to 40%, and male accounts for 13%；It arrives
The year two thousand fifty, hip fracture number will increase to 6,300,000 people by 1,700,000 people of nineteen ninety.According to statistics, osteoporosis influences 2800
Ten thousand Americans, every year because the medical fee and economic loss of osteoporosis and relevant issues reach hundred million dollars of 60-100.There are about 50 for Britain
Ten thousand patients with osteoporosis；Annual 50 years old European or more resident has 1/8 to fracture because of osteoporosis, 414000 generations
Osteoporosis Hip Fracture；It is expected that European osteoporosis treatment related medical expense will increase by 1 times in 50 years.
It is reported that China has become the most country of global patients with osteoporosis, seriousness is only second to angiocarpy
Disease.National extensive epidemiological survey is also shown, and domestic osteoporosis total prevalence rate is 12.4%, the elderly's illness
Ratio is more than more than half, and wherein incidence of fracture is close to 33%.Expect the year two thousand twenty, China's osteoporosis and Low BMD patient
Number will be increased to 2.8 hundred million.
Human parathyroid hormone (parathyroid hormone, parathyroid hormone, PTH) is to maintain body blood calcium flat
One of important hormone of weighing apparatus, basic physiological effect be by increasing osteoblast and osteoclast number, promote bone resorption and
Growth.A large amount of clinical application research show that PTH is one of the most important drug of current treatment osteoporosis.Lilly in 2002
Company(Chemical name: Teriparatide injection) it is approved listing by U.S. FDA, 2011
In March in year, ratified by State Food and Drug Administration in Discussion on Chinese Listed, for treating the post menopausal female of severe osteoporosis
Property patient.The use of recombinant human parathyroid hormone will provide completely new therapeutic scheme to Chinese patients with osteoporosis, help
Improve the quality of life of patient, it helps improve the treatment level of China's osteoporosis, wide market.But it is existing
Its stability of recombinant human parathyroid hormone is bad, is easy to cause the variation of pharmaceutical quantities to affect the treatment.
Summary of the invention
The purpose of the present invention is to provide a kind of recombinant human parathyroid hormone preparation, have at room temperature good
Stability, have better safety.
Another object of the present invention is to provide a kind of preparation method of recombinant human parathyroid hormone preparation, preparation sides
Method is simple, and operation is easy, easy to accomplish.
The embodiment of the present invention is achieved in that
A kind of recombinant human parathyroid hormone preparation is the preparation liquid for mixing raw material and water for injection, raw material
Including recombinant human parathyroid hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent,
Wherein, recombinant human parathyroid hormone stoste is 10-500 μ g/ml preparing the concentration in liquid, and buffer is being prepared
Concentration in liquid is 8-30mmol/L, and osmotic pressure regulator is 40-50mg/ml preparing the concentration in liquid, and bacteriostatic agent is being prepared
Concentration in liquid is 2.4-3.3mg/ml.
A kind of preparation method of recombinant human parathyroid hormone preparation comprising by recombinant human parathyroid hormone stoste,
Buffer, osmotic pressure regulator and bacteriostatic agent are uniformly mixed to form preparation liquid with water for injection as raw material, wherein recombined human
Pro PTH liquid is 10-500 μ g/ml preparing the concentration in liquid, and buffer is 8- preparing the concentration in liquid
30mmol/L, osmotic pressure regulator are 40-50mg/ml preparing the concentration in liquid, and bacteriostatic agent is in the concentration prepared in liquid
2.4-3.3mg/ml。
The beneficial effect of the recombinant human parathyroid hormone preparation of the embodiment of the present invention and preparation method thereof is:
By compounding recombined human first shape in water for injection in recombinant human parathyroid hormone preparation provided in this embodiment
Other glandular hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent effectively increase recombinant human parathyroid hormone preparation
Stability, it is with good stability at room temperature, there is better safety.And in the present embodiment, pass through limit
Fixed above-mentioned recombinant human parathyroid hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent are preparing the concentration in liquid, energy
Enough reach optimal stablizing effect.
In addition, the preparation method of recombinant human parathyroid hormone preparation provided in this embodiment, preparation method is simple, behaviour
It is easy, it is easy to accomplish.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is acceleration for stabilization in recombinant human parathyroid hormone preparation stability comparative experiments provided in an embodiment of the present invention
Property investigate protein concentration variation tendency chart；
Fig. 2 is acceleration for stabilization in recombinant human parathyroid hormone preparation stability comparative experiments provided in an embodiment of the present invention
Property investigate main peak trend chart；
Fig. 3 is to stablize for a long time in recombinant human parathyroid hormone preparation stability comparative experiments provided in an embodiment of the present invention
Property investigate protein concentration tendency chart；
Fig. 4 is to contain quantitative change for 25 DEG C in the choice experiment of recombinant human parathyroid hormone preparation pH provided in an embodiment of the present invention
Change tendency chart；
Fig. 5 is that 25 DEG C of main ingredients are pure in the choice experiment of recombinant human parathyroid hormone preparation pH provided in an embodiment of the present invention
Spend trend chart；
Fig. 6 is to contain quantitative change for 37 DEG C in the choice experiment of recombinant human parathyroid hormone preparation pH provided in an embodiment of the present invention
Change tendency chart；
Fig. 7 is that 37 DEG C of main ingredients are pure in the choice experiment of recombinant human parathyroid hormone preparation pH provided in an embodiment of the present invention
Spend trend chart.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Recombinant human parathyroid hormone preparation of the embodiment of the present invention and preparation method thereof is specifically described below.
A kind of recombinant human parathyroid hormone preparation is the preparation liquid for mixing raw material and water for injection, raw material
Including recombinant human parathyroid hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent.
Recombinant human parathyroid hormone stoste can stimulate bon e formation and bone resorption, can reduce the hair of postmenopausal women's fracture
Raw rate, continuous infusion can lead to parathormone PTH concentration and persistently increase, therefore than only causing serum parathyroid hormone PTH dense
It is stronger to spend the bone resorption that the of short duration daily injection method increased generates.In addition, this medicine does not inhibit adenosine diphosphate (ADP) ways of regeneration
Or the platelet aggregation reaction of collagen-induced approach.In the present embodiment, using recombinant human parathyroid hormone stoste as main ingredient,
Osteoporosis can effectively be treated.
Recombinant human parathyroid hormone is voluntarily prepared by Beijing Bokangjian Gene Technology Co., Ltd, passes through fully synthetic recombination
Target gene is integrated into expression plasmid pET32a (+) and converts Escherichia coli by the gene order of human parathyroid hormone
BL21 (DE3) pLysS, obtaining high efficient expression by screening can be used for the recombinant human parathyroid hormone engineered strain of industrialization,
Engineered strain is cultivated and the thallus containing destination protein is collected by centrifugation after inducing expression, and thallus is crushed, after purification, is obtained high-purity
The recombinant human parathyroid hormone of degree.
Wherein, recombinant human parathyroid hormone stoste is 10-500 μ g/ml, preferably 50-300 preparing the concentration in liquid
μ g/ml, more preferably 250 μ g/ml.Through inventor the study found that recombinant human parathyroid hormone stoste is dense in preparation liquid
When degree is in above range value, the optimal stability of recombinant human parathyroid hormone preparation can deposit when concentration is lower than 10 μ g/ml
The bad stability the problem of, when the concentration of recombinant human parathyroid hormone stoste is more than 5.0mg/ml, can exist the rate of recovery compared with
Low problem.
Further, in the present embodiment, while recombinant human parathyroid hormone stoste is as main ingredient, buffering has been compounded
Liquid, osmotic pressure regulator and bacteriostatic agent are as auxiliary material.
Wherein, buffer can effectively hinder the variation of pH value of solution, and recombinant human parathyroid hormone preparation is made to be in optimal
In pH value.Buffer is 8-30mmol/L, preferably 8-16mmol/L, more preferably 8mmol/L preparing the concentration in liquid.Through
Inventor the study found that buffer prepare liquid in concentration in above range value when, recombinant human parathyroid hormone preparation
Optimal stability, concentration be lower than 25 μ g/ml, can there are problems that bad stability, when recombinant human parathyroid hormone stoste
Concentration be more than 5.0mg/ml, can there is a problem of that the rate of recovery is lower.Specific in the present embodiment, buffer may be vinegar
One of acid-sodium-acetate buffer, citric acid-sodium citrate buffer solution or citrate-phosphate disodium hydrogen buffer.
In addition it is also necessary to illustrate, the Acetic acid-sodium acetate buffer in the present embodiment is by glacial acetic acid and three hydration vinegar
Sour sodium composition, in Acetic acid-sodium acetate buffer, the concentration ratio of glacial acetic acid and sodium acetate trihydrate is 5-6:1.In the concentration ratio
The buffer for the Acetic acid-sodium acetate buffer being formulated under example is more stable.Likewise, citric acid-sodium citrate buffer solution
In, the concentration of citric acid and sodium citrate is than being also 5-6:1.
Osmotic pressure regulator is conducive to adjust the osmotic pressure for preparing liquid, keeps osmotic pressure to be in zone of reasonableness, avoided
Hemolytic reaction caused by low and excessively high caused cell dehydration reaction etc..In the present embodiment, osmotic pressure regulator includes sweet
Reveal at least one of alcohol injection, glycine, methionine, glucose and sucrose.It that is to say, osmotic pressure regulator can be with
It, can also be selected from formula mannitol injection liquid, sweet selected from single formula mannitol injection liquid, glycine, methionine, dextrose and saccharose
Propylhomoserin, methionine, in dextrose and saccharose any 2-5 person combination.
Wherein, mannitol is monosaccharide in formula mannitol injection liquid, is not metabolized in vivo, in renal tubule after glomerular filtration
It is inside very few by reabsorption, play the role of osmotic diuresis.Be conducive to improve plasma osmolarity, moisture in tissue is caused to enter blood vessel
It is interior, to mitigate tissue edema, reduce intraocular pressure, intracranial pressure and cerebrospinal fluid capacity and its pressure.Likewise, glycine, first sulphur
Propylhomoserin, dextrose and saccharose can play the role of certain osmotic pressure and adjust.By applicants have found that, select mannitol
For injection as osmotic pressure regulator, osmotic effect is good.
In the present embodiment, osmotic pressure regulator prepare liquid in concentration be 40-50mg/ml, preferably 42-47mg/ml,
More preferably 45mg/ml.Through inventor the study found that osmotic pressure regulator is preparing the concentration in liquid in above range value
When, the optimal stability of recombinant human parathyroid hormone preparation, concentration is lower than 35mg/ml, can there are problems that bad stability,
When the concentration of osmotic pressure regulator is greater than 50mg/ml, there can be albumen aggregation, the problem of insoluble granule increases.
Bacteriostatic agent can effectively prevent bacteria breed, be conducive to the stabilization formulations for keeping recombinant human parathyroid hormone preparation
Stability, and then extend its resting period.In the present embodiment, bacteriostatic agent is 2.4-3.3mg/ml preparing the concentration in liquid；
Preferably 2.6-3mg/ml, more preferably 3mg/ml；Bacteriostatic agent includes at least one of metacresol and benzyl alcohol.Through inventor
The study found that when concentration of the bacteriostatic agent in preparation liquid is in above range value, the stabilization of recombinant human parathyroid hormone preparation
Property it is best, concentration be lower than 2.4mg/ml, can there is a problem of that inhibitory effect is not up to standard, when the concentration of bacteriostatic agent is more than 3.5mg/ml
When, can there are problems that generating toxic side effect.
Wherein, 1% m-cresol solution can prepare by the following method (amount of preparation 1.0L): between being measured using glass pipette
Cresols is placed in clean beaker, adds water to be settled to 1000ml, be stirred using magnetic stirring apparatus, and the time answers >=
20min is to being completely dissolved.It is filtered operation using 0.2 μm of filter, it is stand-by that filtered fluid deposits in the blue lid bottle of sterilizing.
In addition, the pH value and record of liquid are prepared in detection when preparing above-mentioned preparation liquid, if meeting pH is 3.5-5.0, such as
Fruit is unsatisfactory for, then is adjusted using 1.0mol/L sodium hydroxide or 1.0mol/L hydrochloric acid and prepare liquid pH to 3.5-5.0, hydroxide is added
It should the slowly simultaneously light and slow stirring when being added when sodium or hydrochloric acid.Preferably, pH 3.5-4.5, more preferably 4.0.
The present embodiment additionally provides a kind of preparation method of recombinant human parathyroid hormone preparation comprising by recombined human first
Glandular hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent are uniformly mixed to be formed and match as raw material with water for injection by shape
Liquid processed.Specifically, comprising the following steps:
S1: pretreatment of raw material
Before recombinant human parathyroid hormone stoste is added, it is placed on by the recombined human first shape stored at -25~-15 DEG C
Glandular hormone stoste is placed in the water-bath that temperature is 20-30 DEG C, is shaken to recombinant human parathyroid hormone stoste and is dissolved.
Recombinant human parathyroid hormone stoste is placed at -25~-15 DEG C and is stored, is conducive to keep its stability, then
Before feeding intake, warming-in-water is carried out to it, is made it dissolve, is conducive to subsequent and other raw materials mixing.
S2: it feeds intake in order.
Buffer, osmotic pressure regulator, bacteriostatic agent and recombinant human parathyroid hormone are sequentially added into water for injection
Stoste is uniformly mixed.
After buffer is added, liquid agitation in bottle after bacteriostatic agent is added, is utilized to being completely dissolved using glass bar
IKA overhead type stirrer is stirred, and adjusting revolving speed is 100-150 turns/min to the liquid agitation 6-10min in bottle；Adding
After entering recombinant human parathyroid hormone stoste, continue to be stirred the liquid in bottle, mixing time 6-10min, stirring turns
Speed is that 60-90 turns/min.Liquid is to prepare liquid in bottle after the completion of this step.
In the present embodiment, liquid is stirred by the mixing speed of different rotating speeds twice, can guarantee the mixed of liquid
Uniformity is closed, meanwhile, it when stirring for the first time, is stirred using faster speed 100-150 turns/min, is conducive to accelerate solution
Uniform mixing, and stir for the second time, the slower speed 60-90 of use turns/min is stirred, and tracing it to its cause is because being added
Recombinant human parathyroid hormone stoste, stirring at low speed can effectively avoid recombinant human parathyroid hormone stoste from generating foam,
And then keep preparing the uniform stabilization of liquid.
In stock, need to meet recombinant human parathyroid hormone stoste preparing the concentration in liquid is 10-500 μ g/ml, is delayed
Fliud flushing is 8-30mmol/L preparing the concentration in liquid, and osmotic pressure regulator is 40-50mg/ml, suppression preparing the concentration in liquid
Microbial inoculum is 2.4-3.3mg/ml preparing the concentration in liquid.
S3: pH is adjusted.
The pH value and record of liquid are prepared in detection, if meeting pH is 3.5-5.0, if conditions are not met, then using 1.0mol/L
Sodium hydroxide or 1.0mol/L hydrochloric acid adjusting preparation liquid pH to 3.5-5.0, should slowly when sodium hydroxide or hydrochloric acid is added and side adds
Enter the light and slow stirring in side.Preferably, pH 3.5-4.5, more preferably 4.0.
S4: aseptic filtration.
It further include that aseptic filtration, filter pressure 0.01-0.1Mpa, filtering are carried out to preparation liquid after preparation liquid is made
Time is 5-20min.Through inventor the study found that using being filtered under certain pressure, effectively preparation liquid can be removed
Bacterium filtering, filter effect more, more clarify, and stability is further ensured by filtered preparation liquid.
S5: filling and jump a queue.
Filling and stopper-adding, packing, separation container be sterile 3ml clamped bottle, filling loading amount be 2.55ml/ branch, packing precision ±
1%.Filling speed is 50-100 branch/min.
S6: lid is rolled.
Rolling the every 10min of lid operation should use continuous 12 product examination of 3 fingering extraction roll lid situation, and rolling lid should fasten.
Sample acquired in pouring process should individually be carried out rolling lid.
S7: lamp inspection.
Operator's eyesight should should carry out full mesh to the product that lid is completed is rolled 4.9 or more (correct defects of vision 5.0 or more)
Inspection, lamp inspection station illumination should meet the requirement of 1000-1500lx, and lamp inspection recall rate answers >=98%.
S8: packaging
It is transported in 2-8 DEG C of freezer and keeps in after note label, detection.
With reference to embodiments to recombinant human parathyroid hormone preparation of the invention and preparation method thereof further progress
It illustrates.
Embodiment 1
Present embodiments provide a kind of recombinant human parathyroid hormone preparation and preparation method thereof.
Raw material includes: recombinant human parathyroid hormone stoste, Acetic acid-sodium acetate, formula mannitol injection liquid, metacresol and note
It penetrates and uses water.Before being on the waiting list recombinant human parathyroid hormone stoste, it is placed on by the recombined human first shape stored at -25~-15 DEG C
Glandular hormone stoste is placed in the water-bath that temperature is 20-30 DEG C, is shaken to recombinant human parathyroid hormone stoste and is dissolved.
Preparation method: Acetic acid-sodium acetate being added into water for injection, using glass bar by liquid agitation in bottle to completely it is molten
Solution, is subsequently added into formula mannitol injection liquid, continues to stir, and metacresol is then added, and the revolving speed for adjusting IKA overhead type stirrer is
100 turns/min is to the liquid agitation 6min in bottle；Then recombinant human parathyroid hormone stoste is added, continues to the liquid in bottle
It is stirred, mixing time 6min, speed of agitator is 80 turns/min, is uniformly mixed.Then 1.0mol/L sodium hydroxide is used
Or 1.0mol/L salt acid for adjusting pH prepares liquid to 4, carries out aseptic filtration 10min in the case where filter pressure is 0.05Mpa.
In preparation liquid obtained, the concentration of recombinant human parathyroid hormone stoste is 250 μ g/ml；Acetic acid-sodium acetate
Concentration is 8mmol/L；The concentration of formula mannitol injection liquid is 45mg/ml, the concentration of metacresol is 3mg/ml.
Embodiment 2-5
Embodiment 2-5 provides a kind of recombinant human parathyroid hormone preparation and preparation method thereof.Its raw material and preparation side
Method is same as Example 1, and difference is: in the present embodiment, raw material is preparing the concentration difference in liquid:
In example 2, in preparation liquid obtained, the concentration of recombinant human parathyroid hormone stoste is 10 μ g/ml；Lemon
Acid-sodium citrate buffer solution concentration is 30mmol/L；The concentration of formula mannitol injection liquid is 40mg/ml, the concentration of metacresol is
2.4mg/ml, the pH value for preparing liquid is 3.5.
In embodiment 3, in preparation liquid obtained, the concentration of recombinant human parathyroid hormone stoste is 50 μ g/ml；Lemon
Acid-sodium citrate buffer solution concentration is 8mmol/L；The concentration of formula mannitol injection liquid is 42mg/ml, the concentration of metacresol is
2.6mg/ml, preparing liquid pH value is 5.
In example 4, in preparation liquid obtained, the concentration of recombinant human parathyroid hormone stoste is 300 μ g/ml；Vinegar
Acid-sodium-acetate buffer concentration is 16mmol/L；The concentration of formula mannitol injection liquid is 47mg/ml, the concentration of metacresol is
2.8mg/ml, preparing liquid pH value is 4.5.
In embodiment 5, in preparation liquid obtained, the concentration of recombinant human parathyroid hormone stoste is 500 μ g/ml；Lemon
Lemon acid-disodium hydrogen phosphate buffer concentration is 10mmol/L；The concentration of formula mannitol injection liquid be 50mg/ml, metacresol it is dense
Degree is 3.3mg/ml, and preparing liquid pH value is 3.5.
Embodiment 6-10
Embodiment 6-10 provides a kind of recombinant human parathyroid hormone preparation and preparation method thereof.Its raw material and preparation side
Method is same as Example 1, and difference is: in the present embodiment, raw material is different in the selection of osmotic pressure regulator and bacteriostatic agent:
In embodiment 6, osmotic pressure regulator is glycine, and bacteriostatic agent is benzyl alcohol.
In embodiment 7, osmotic pressure regulator is methionine, and bacteriostatic agent is metacresol and benzyl alcohol, metacresol and benzene first
The mass ratio of alcohol is 1:2.
In embodiment 8, osmotic pressure regulator is the mixture of sodium chloride and glucose, and bacteriostatic agent is benzyl alcohol.
In embodiment 9, osmotic pressure regulator is the mixing of glycine, methionine, sodium chloride, glucose and sucrose
Object, bacteriostatic agent are metacresol.
In embodiment 10, osmotic pressure regulator be formula mannitol injection liquid, glycine, methionine, sodium chloride, glucose with
And the mixture of sucrose, bacteriostatic agent are metacresol and benzyl alcohol, the mass ratio of metacresol and benzyl alcohol is 1:2.
One, stability contrast is tested
A: embodiment 1；
B: comparative example 1, the bacteriostatic agent in embodiment 1 are metacresol, and metacresol is replaced with conventional bacteriostatic agent in comparative example 1
Methyl hydroxybenzoate sodium；
C: comparative example 2, buffer is Acetic acid-sodium acetate buffer in embodiment 1, delays Acetic acid-sodium acetate in comparative example 2
Fliud flushing replaces with citrate-phosphate disodium hydrogen buffer.
D: comparative example 3, multiple safe injection difficult to understand.
(1) accelerated stability is tested
(25 ± 2 DEG C) of the alternative prescription accelerated stability of recombinant human parathyroid hormone injection experiments, institute as shown in Figure 1, Figure 2
Show, embodiment 1 (A) is from the variation of T0-3M appearance, protein concentration and main peak % etc. compared to comparative example 1 (B) and comparative example 2
(C) smaller, compared with comparative example 3 (D) multiple safe Austria, variation tendency is similar.
(2) long-time stability are tested
(5 ± 3 DEG C) of the alternative prescription long-time stability of recombinant human parathyroid hormone injection experiments, as shown in figure 3, implementing
(A) protein concentration of example 1 reduces degree minimum from T0-3M, and comparative example 1 (B) fall is maximum, and comparative example 1 is substantially better than comparison
Example 1 and 2.
The master under the conditions of multigelation, accelerated stability (25 ± 2 DEG C), long-time stability (5 ± 3 DEG C) is comprehensively compared
Number under the conditions of peak %, main ingredient related impurities (RRT0.85, RRT0.90, RRT0.98, RRT1.08, RRT1.13), appearance, pH
According to the obviously preferred comparative example 1 of embodiment 1 provided in this embodiment, comparative example 2 and comparative example 3, stablizing effect is good.
Two, the choice experiment of pH
The buffer solution system of different pH is prepared, pH is respectively 4,5,6.Then test accelerates to contain under the conditions of 25 DEG C with 37 DEG C
Measure the trend of purity variation.Please refer to Fig. 4-7.
A1: embodiment 1, pH value 4.
Aa: comparative example 4, pH value 5.
Ab: to you 5, pH value 6.
It can be seen that the pH4.0 buffer prescription of the offer of embodiment 1 in 25 DEG C and 37 DEG C according to Fig. 4, Fig. 5, Fig. 6 and Fig. 7
Content purity is stablized compared with the pH5.0 of the offer of comparative example 4 and 5 with 6.0 prescriptions in accelerator.
In conclusion by compounding weight in water for injection in recombinant human parathyroid hormone preparation provided in this embodiment
Group human parathyroid hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent, effectively increase recombinant human parathyroid hormone
The stability of plain preparation, it is with good stability at room temperature, there is better safety.And the present embodiment
In, by limiting above-mentioned recombinant human parathyroid hormone stoste, buffer, osmotic pressure regulator and bacteriostatic agent in preparing liquid
Concentration, optimal stablizing effect can be reached.
In addition, the preparation method of recombinant human parathyroid hormone preparation provided in this embodiment, preparation method is simple, behaviour
It is easy, it is easy to accomplish.
These are only the preferred embodiment of the present invention, is not intended to restrict the invention, for those skilled in the art
For member, the invention may be variously modified and varied.All within the spirits and principles of the present invention, it is made it is any modification,
Equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (3)
1. a kind of preparation method of recombinant human parathyroid hormone preparation, which is characterized in that it include into water for injection successively
Concentration is added than first between the buffer, osmotic pressure regulator mannitol, bacteriostatic agent of glacial acetic acid and sodium acetate trihydrate for 5-6:1
Phenol, 100-150 turn/min stirring 6-10min；Recombinant human parathyroid hormone stoste is added, 60-90 turns/min stirring 6-
10min is uniformly mixed to be formed and prepares liquid, wherein concentration of the recombinant human parathyroid hormone stoste in the preparation liquid
For 250 μ g/ml, concentration of the buffer in the preparation liquid is 8mmol/L, and the osmotic pressure regulator is in the preparation
Concentration in liquid is 45mg/ml, and concentration of the bacteriostatic agent in the preparation liquid is 3mg/ml；The pH of the preparation liquid is
4.0。
2. the preparation method of recombinant human parathyroid hormone preparation according to claim 1, which is characterized in that in obtained institute
It states after preparing liquid, further includes that aseptic filtration, filter pressure 0.01-0.1Mpa, filtration time 5- are carried out to the preparation liquid
20min。
3. the preparation method of recombinant human parathyroid hormone preparation according to claim 1, which is characterized in that institute is being added
Before stating recombinant human parathyroid hormone stoste, it is placed on the recombinant human parathyroid hormone stored at -25 ~ -15 DEG C original
Liquid is placed in the water-bath that temperature is 20-30 DEG C, is shaken to the recombinant human parathyroid hormone stoste and is dissolved.
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