CN1089249C - Biocompatible somatotropin solutions - Google Patents
Biocompatible somatotropin solutions Download PDFInfo
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- CN1089249C CN1089249C CN93104030A CN93104030A CN1089249C CN 1089249 C CN1089249 C CN 1089249C CN 93104030 A CN93104030 A CN 93104030A CN 93104030 A CN93104030 A CN 93104030A CN 1089249 C CN1089249 C CN 1089249C
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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
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- 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
- A61K38/27—Growth hormone [GH] (Somatotropin)
-
- 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
-
- 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
Abstract
Somatotropin solutions which remain clear for extended periods and which remain clear when subjected to mechanical agitation are prepared by mixing a lyophilized somatotropin composition containing about 1 part somatotropin per 2 to 8 parts arginine HC1 on a weight basis, wherein the pH of the composition prior to lyophilization was about 7.2 to about 8.5, and a biocompatible diluent which comprises EDTA and a nonionic surfactant. If the pH of the somatotropin composition prior to lyophilization was between 7.2 and 7.8, the diluent further comprises a buffer; otherwise the diluent optionally further can comprise a buffer or nonbuffering agent.
Description
The present invention relates to a kind of new soln of growth hormone and prepare required equipment.It is thorough clearly that this solution can keep in a long time, also can keep thorough clearly under mechanical oscillation.
Growth hormone (somatotropins), that is growth hormones are by the excretory polypeptide hormone of multiple animal pituitary gland.This parahormone has multiple therapeutic use, and its synthetic can be used for treating human hypophysis cerebri defective, gastrointestinal hemorrhage and the agglutination that quickens fracture, contusion and other wound.By different application methods and injection, can also promote the production of animal flesh, breast.(referring to:
E.J.?Turman，?″Some?Effects?ofPituitary?Anterior?Growth?Factor″?Thesis：PurdueUniversity，April，1953；L.J.Machlin，J.Anim.Sci.35：794-800(1972)；T.R.Kasser?et?al.，J.?Anim.?Sci.53：420-426；L.J.Machlin，J.Dairy?Sci.?56：575-580(1973)).
The processing of this proteinoid of growth hormone and use usually need be used its solution, yet, thereby dissolved growth hormone such as protein can be adsorbed the generation secondary response by hydrophobic interfaces.As " degeneration ".Promptly Xi Fu growth hormone molecular shape changes.In addition, the aggregation of the growth hormone molecule of absorption also can produce solvable or insoluble polymerized form.The example of this aggregation performance is, when stirring and during the aqueous solution of oscillating growth hormone, and the solution muddiness, or growth hormone loses biological activity.(referring to people such as A.F.Henson, J.ColloidInterface Sci.32:162 (1970))
After growth hormone was precipitated out from solution, the protein that is settled out just can not reuse, therefore, need a kind of can be muddy after stirring vibration, store longer a period of time and still can keep thorough clearly solution.
It is thorough clearly that growth hormone solution disclosed by the invention can keep for a long time, and also can not produce muddiness and precipitation under mechanical oscillation.This solution comprises cryodesiccated somatotropin composition, and the every 2-8 part of said composition arginine hydrochloride contains about 1 part of growth hormone (weight ratio), and the lyophilization up till now pH value of compositions is dissolved in the diluent between 7.2-8.5.If the pH value of somatotropin composition is lower than 7.8 before the lyophilization, diluent contains a kind of non-ionic surface activator of ethylenediaminetetraacetic acid (EDTA) and place's buffer agent; If the pH value of somatotropin composition is 7.8 or higher before the lyophilization, then this diluent comprises EDTA and a kind of non-ionic surface activator, and sending out needs also can comprise a kind of suitable reducing or non-buffer medium.Though two media all can use,, just can not add buffer agent again if add non-buffer medium.
The present invention also provides the equipment of this solution of preparation, comprise a bottle that cryodesiccated somatotropin composition (every 1 part of auxin of 2-8 part arginine hydrochloride (weight ratio)) is housed, comprise that also another is equipped with the bottle of the diluent of biocompatible, interior dress EDTA and a kind of non-ionic surface activator.In addition, as previously mentioned, according to the pH value of somatotropin composition before the lyophilization, this diluent also can contain a kind of buffer agent or non-buffer medium.If the pH value of somatotropin composition also should contain a kind of buffer agent less than 7.8 in the bottle before the lyophilization.Be equal to or greater than 7.8 as PH, contain a kind of buffer agent or non-buffer medium in the bottle alternatively.Can shake the gained mixture with in the somatotropin composition after this diluent adding lyophilization, make the growth hormone dissolving.
The invention provides can keep thorough clearly for a long time and at mechanical oscillation, shake under stirring with eddy current still haze-free and can not produce sedimentary growth hormone solution.The so-called growth hormone solution that can keep " thorough clearly " is that growth hormone is dissolved in non-setting solution behind the diluent.This solution was being stored several days, generally still kept thorough clearly after at least five days, and this solution generally kept thorough clearly on 5 under 25 ℃.High slightly as storage temperature, it is thorough clearly that this solution still can keep, and just the holding time may shorten.A kind ofly keep thorough clearly solution, its at the apparent absorbance (0.0360) at 360nm place less than 0.10.
The brilliant people of this growth hormone solution contain cryodesiccated somatotropin composition, and the pH value before the lyophilization is at least 7.2, generally between 7.2-8.5.If the pH value of growth hormone synthetic is less than 7.8 before the lyophilization, diluent then comprises EDTA, and a kind of non-ionic surface activator and a kind of suitable reducing are to stablize final growth hormone solution.If the pH value of somatotropin composition is more than or equal to 7.8 before the lyophilization, diluent then comprises EDTA, a kind of non-ionic surface activator and comprise a kind of buffer agent alternatively or non-buffer medium.Though two media all can use,, then had better not add buffer agent again if add non-buffer medium.If the pH value of somatotropin composition is more than or equal to 7.8 before the lyophilization, buffer agent and non-buffer medium are all optional, but the isotonicity of wherein any equal scalable and maintenance gained growth hormone solution is in order to avoid it becomes hyperosmotic solution, and this is essential in animal injection growth hormone solution.Suitably the selection of additive comprises for example price (FDA shows among the GRASList) based on many factors, the autoclaving ability, and chemical stability does not have interaction and biocompatible with growth hormone.
Being prepared as of cryodesiccated somatotropin composition is dissolved in arginine hydrochloride in the growth hormone, more than the pH value to 7.2 of regulator solution, be advisable with 7.2-8.5, remove insoluble matter with filtration and centrifuging then, with the standardization program lyophilization of growth hormone and arginine hydrochloride solution usefulness known technology.
Cryodesiccated solution generally includes, and every 1 part of growth hormone of 2-8 part arginine hydrochloride (weight ratio) is preferably 1 part of growth hormone of per 3 parts of arginine hydrochloride.Therefore, this solution preferably includes 10-150mg/ml growth hormone and 30-450mg/ml arginine hydrochloride, and final pH value is not less than 7.2.Optimumly be, cryodesiccated solution comprises 30mg/ml growth hormone and 90mg/ml arginine hydrochloride, and its final pH value is between 7.2-8.5.
Growth hormone among the present invention in the cryodesiccated somatotropin composition can be any class growth hormone, comprises the growth hormone of natural or synthetic cattle, pig, people, bird, sheep or horse, the growth hormone of the most handy pig.Noun " growth hormone " is meant the derivant that comprises complete natural hormone or synthetic growth hormone and have the growth promotion ability altogether herein.Derivant comprises the part of biologically active or the analog of this type of polypeptide hormone.δ-7 pig growth hormone (delta 7 porcine somatotropin) for example, it has the amino identical with pig growth hormone arranges, and lacks first seven amino acid (seeing EuropeanPatent Application Publication No.0 104 920 to BiogenN.V.) than ripe, complete pig growth hormone. and term " biological activity " is meant that this polypeptide is used for behind the live body its bioprocess being had remarkable influence herein.
Used growth hormone can be the growth hormone of melts combine among the present invention.The growth hormone of melts combine produces by add transition metal salt in containing the aqueous solution of growth hormone.These transition metal salts and growth hormone produce insoluble complex, are precipitated out from solution.The growth hormone of melts combine contains growth hormone molecule and metal ion, as Zn
+ 2, Cu
+ 2, Co
+ 2, Mn
+ 2Fe
+ 2And Fe
+ 3, wherein the nitrogen-atoms on some amino acid residue links to each other with coordinate bond in metal ion and the growth hormone molecule.The growth hormone of melts combine can be used as the raw material of the cryodesiccated somatotropin composition of preparation.Although contain metal in the cryodesiccated growth hormone synthetic, in case through lyophilization, metal just no longer links to each other with growth hormone.When being used for organism, the transition metal in the product there is no any significant adverse effect to the biological activity of growth hormone.
Growth hormone solution of the present invention can prepare by cryodesiccated somatotropin composition is dissolved in the diluent, and the final solution pH value is at least 7.2, is generally about 7.2-8.5, is preferably between the 7.2-8.2.If the pH value of solution is greater than 8.5, protein will be degraded.Usually, the cryodesiccated somatotropin composition of the about 0.5-40mg of dissolving in the 1ml diluent, the ultimate density of growth hormone solution is advisable with every milliliter of cryodesiccated production hormonal composition of diluent 10-30mg, is preferably every milli and opens the cryodesiccated somatotropin composition of the about 20mg of diluent.The weight of cryodesiccated somatotropin composition is wherein contained arginine hydrochloride and growth hormone weight sum.
If the pH value before the somatotropin composition lyophilization of preparation growth hormone solution is less than 7.8, then diluent contains EDTA, a kind of non-ionic surface activator and a kind of buffer agent.On the other hand, if the pH value of somatotropin composition more than or equal to 7.8, then diluent contains EDTA, a kind of non-ionic surface activator and contain a kind of buffer agent alternatively or non-buffer medium.If diluent contains buffer agent, then the pH value of final growth hormone solution will be consistent with the pH value of buffer, if but the diluent right and wrong are buffered, and then the pH value of somatotropin composition will determine the pH value of final growth hormone solution.
If the solution pH value of final growth hormone is between about 7.2-7.6, the concentration of used EDTA should be at least 1.5mM in the diluent, is preferably 1.5mM-10mM.If the pH value of final growth hormone solution is greater than 7.6, then the concentration of used EDTA is at least 1.0mM in the diluent, and is the most suitable between 1.0mM to 10mM.
Be applicable to that the surfactant in the diluent comprises: polyoxyethylene-23-Laurel ether (background of cloth agent Brij35), Tween 80, polyoxyethylene-20-cetyl ether (background of cloth agent Brij58) and other have the polyoxyethylene non-ionic surface activator of similar hydrophilic/hydrophobic balance (HLB).Prior art shows that this type of non-ionic surface activator can stablize and keep the activity of purifying enzyme.(see T.Kitani et al.Eur.J.Biochem.119:177-181 (1981); M.Pritchard et al., Biochem.Biophys.Res.Commun.100:1597-1603 (1981); Seely et al.Biochemistry, Vol.21, No.14,3394-3399 (1982)).Usually, the concentration range of used surfactant is for being about 0.08%-2.0%.If surfactant Brij35, then the concentration of Brij is at least 0.08%, is preferably between the 0.1%-0.2%.If surfactant is with Tween 80 or Brij58, then its concentration is advisable with 0.1%-1.0%.
If the pH value of growth hormone is more than or equal to 7.2 less than 7.8 before the lyophilization, then the buffer agent in the diluent can increase the stability of growth hormone solution.Buffer agent can be hydrochloric acid Tris (Tris HCl), phosphate or other neutral compatible with receptor PH buffer agent.Usually, the concentration range of adding buffer agent is about 0.2-0.5M.If make buffer agent with Tris HCl, the concentration of Tris HCl is at least 0.2M, is preferably 0.2M-0.3M.
If the pH value of growth hormone synthetic can add foregoing buffer agent or non-buffer medium more than or equal to 7.8 in the diluent as required before the lyophilization, to regulate or to keep the isotonicity of growth solution.Non-buffer medium also should be compatible with receptor.The non-buffer medium of this class comprises sucrose, trehalose or sodium chloride.If use non-buffer medium.Its concentration should be 0.05M-0.5M.If non-buffer medium is 0.1-0.3M with sucrose or trehalose, suitable concentration range.As non-buffer medium sodium chloride, concentration is advisable with 0.05M-0.15M.
Prepare equipment of the present invention and comprise that the bottle that aforementioned cryodesiccated somatotropin composition is housed is equipped with the bottle of aforementioned diluent with another.Diluent is injected cryodesiccated growth hormone compound vibration make the growth hormone dissolving.
Growth hormone solution of the present invention can be used for further processing or directly is applied to animal body.This solution at room temperature (is generally 25 ℃) can preserve at least 5 days haze-free.This solution also can be stored under the higher slightly temperature, although its period of storage may shorten.This solution can not be settled out long-living hormone yet under mechanical oscillations such as vibration, stirring.Growth hormone in the solution can keep biological activity, and technology is applied to animal body to promote growth routinely.
The also available following examples explanation of the present invention, but these embodiment do not constitute limiting the scope of the invention.
Embodiment
The clear thorough degree of solution is measured with measuring apparent optical density (O.D.360) under visual method and/or the 360nm in following examples, and auxin does not have interior absorption under this wavelength.Growth hormone solution is contained in the sample cuvette, O.D.360 Shimadzu UVU 160 spectrophotometric determinations.The diluent that does not contain growth hormone in contrast.Quartz cuvette is all used in all experiments." thorough clearly " is meant that O.D.360 is less than 0.10 solution among the embodiment.
Embodiment 1
Close pig growth hormone (PST) according to No. 83300803.9 disclosed method preparation 32 gram zinc of european patent application, itself and 90 gram arginine hydrochloride are dissolved in 1 liter of sterilized water, with hydrochloric acid or sodium hydrate aqueous solution pH value is transferred to 7.8, with poly-difluoroethylene membrane filtration, remove insoluble matter, then this transparency liquid is carried out lyophilization.Product (called after " cryodesiccated pig growth hormone (PST) compositions) contains the about 30 gram PST of per 90 gram arginine hydrochloride, and filter process loses among the former 33 gram PST 10% insoluble PST.
Embodiment 2
Containing 0.639g PST and 21.6ml pH value is 9.8, and the non-zinc of the sodium carbonate buffer of 0.46M closes and adds 1.92 gram solid arginine hydrochloride in the PST solution, stirs, and makes the arginine dissolving, drips the 1M sodium hydroxide solution and regulates pH value to 7.8.Generate solution and contain have an appointment 30mg/mlPST and 90mg/ml arginine.Move in the bottle of three 50ml with the aliquot (7.2ml) of pipet, be refrigerated to-80 ℃ and carry out lyophilization then solution.
Cryodesiccated PST compositions is soluble in 200mM Tris HCl, 2mM EDTA, and among the 0.15%Brij35 (PH is 7.8), concentration is 20mg/ml (dissolution time was less than 3 minutes).It is thorough clearly observing generation solution with visual method.
Embodiment 3
Prepare following solution:
1.?0.2M?Tris?HCl，2mM?EDTA，0.15％Brij?35，pH?7.8
2.?″ ″ ″ ″ ″ ″ ″ ，pH?7.4
3.2mM?EDTA，0.15％Brij?35，pH?7.8
The PST (Zn-PST) that zinc is closed, nonmetal bonded PST (non-Zn PST) and cryodesiccated PST compositions (with the method preparation of embodiment 1) are dissolved in above solution respectively, vibrate 2 minutes (concentration is the cryodesiccated PST compositions of 5mg/ml PST or 20mg/ml).The clear thorough degree of gained solution after 2 minutes of vibrating is measured with visual method with in the method that 360nm measures turbidity.As shown in table 1, have only cryodesiccated PST compositions can obtain thorough clearly solution with this understanding.The result is as shown in table 1:
" #3 0.019 for table 1PST solution O D360Zn-pST #1 0.522 " #2 0.504 " #3 0.381non-Zn pST #1 0.242 " #2 0.347 " #3 0.186 cryodesiccated PST composition " #1 0.019 " #2 0.019
Embodiment 4
The diluent that preparation is following: 1. 0.2M 2mM
Tris HCl EDTA PH7.8 contains 0.15% BriJ352 " " " " " 0.125% ".3. ″ ″ ″ ″ ″ 0.10％ ″ 。4. ″ ″ ″ ″ ″ 0.08％ ″ 。5. ″ ″ ″ ″ ″ 0.06％ ″ 。6. ″ ″ ″ ″ ″ 0.04％ ″ 。7. ″ ″ ″ ″ ″ 0.00％ ″ 。
Claim the cryodesiccated PST compositions of 40mg (with the method preparation of embodiment 1) of seven parts of five equilibriums to insert respectively in seven 13 * 100 the test tube, dissolve with the above diluent of 2ml respectively. gained solution stirred 30 seconds with vortex agitator (transferring to 4.5); Measure O.D.360 with visual method and spectra photometric method, its result is as shown in table 2: table 2 different diluent affects dilution numbering performance O.D.3601 clear thorough 0.0402 clear thorough 0.0473 clear thorough 0.0724 clear thorough 0.0925 slightly muddy 0.2576 slightly muddy 0.4847 very muddy 1.708 to restructuring PST mechanical stability
As shown in table 2, cryodesiccated somatotropin composition in the whirling motion agitation and dilution liquid with this understanding, have only contained Brij35 concentration can keep thorough clearly, and contained Brij35 concentration after stirring, whirling motion can not keep thorough clearly less than 0.08% growth hormone solution greater than 0.08% solution.
Embodiment 5
The cryodesiccated PST compositions that claims the 40mg five equilibrium is in the vial of 2 drachms, then with containing 200mM Tris HCl, the diluent dissolving of 0.10%Brij35 and following EDTA concentration and final pH value: 1. 2.0 mM EDTA, pH 7.22. 0.25 mM "; pH 7.43. 0.50 mM ", " 4. 1.00 mM ", " 5. 1.50 mM ", " 6. 2.00 mM ", " 7. 0.25 mM ", pH 7.68. 0.50 mM "; " 9. 1.00 mM ", " 10. 1.50 mM ", " 11. 2.00 mM "; " 12. 0.25 mM "; pH 7.813. 0.50 mM ", " 14. 1.00 mM ", " 15. 1.50 mM ", " 16. 2.00 mM ", "
These samples are placed 25 ℃ of calorstats, and at different time its clear thorough degree that detects by an unaided eye, the result is as shown in table 3
Table 3
Different diluent on restructuring PST stability of solution affect solution # 1 days 5 days 10 days 1 clear thorough clear thorough clear thorough 2 slightly muddiness slightly the clear thorough slightly muddy 10 clear thorough slightly muddy clear thorough 11 clear thorough slightly muddy clear thorough 12 clear thorough muddinesses very muddy 13 clear thorough clear thorough muddy 14 of the clear thorough slightly muddiness of the clear thorough slightly muddy 5 clear thorough clear thorough clear thorough 6 clear thorough clear thorough clear thorough 7 clear thorough muddinesses very muddy 8 of the clear thorough muddiness of muddiness very muddy 3 very muddy 4 very muddy 9 are clear thorough clear thorough slightly muddy 15 clear thorough clear thorough clear thorough 16 clear thorough clear thorough clear thorough as shown in table 3; Contain concentration under this condition greater than 1.5mM EDTA; PH is that growth hormone solution maintenance at least 10 days after cryodesiccated PST synthetic is dissolved in dilution of 7.2-7.6 is clearly thorough. And containing concentration under this condition greater than 1.0mM EDTA, pH value still can keep thorough clearly at least in 5 days greater than 7.6 growth hormone solution after cryodesiccated PST synthetic is dissolved in diluent.
The cryodesiccated PST synthetic that embodiment 6 claims 40mg to be divided into 16 parts is inserted in the bottle of 16 two drachms, adds following diluent 2ml; 1. 300 mM Tris HCl, 0.15% Brij 35,1.5 mM EDTA, pH 7.42. 200 mM Tris HCl, " " " " " 3. 100 mM Tris HCl; " " " " " be 50 mM Tris HCl 4., " " " " " 5. 25 mM Tris HCl; " " " " " be 200 mM Tris HCl 6., " " 2.0 mM EDTA, pH 7.27. " " "; " " 1.5 mM EDTA; " " " "; " " 1.0 mM EDTA; " 9. 10% sucrose, 0.15% Brij 35,2 mM EDTA10., 5% sucrose, " ", " " 11. 1% sucrose, " ", " " 12. 0.2 M NaCl, 0.15% Brij, 35,2 mM EDTA13., 0.1 M NaCl, " ", " " 14. 0.0 M NaCl, " ", " "
With the final pH value of sample after the diluent 9-14 reorganization be 7.8-7.9 (or approximate lyophilization before the last pH value of regulating of PST/ arginine solution).These samples place 25 ℃ of calorstat insulations 6 days, measure its turbidity (O.D.360) result then.As shown in table 4.
Table 4
Different diluent is to influence (O.D.360) behind solution # 6 day of reorganization PST stability of solution
1 0.021
2 0.064
3 0.264
4 0.537
5 0.658
6 0.064
7 0.084
8 0.979
9 0.025
10 0.025
11 0.025
12 0.025
13 ＜0.03
14 ＜0.03
Shown in solution 1-5 in the table 4, contain 0.15%Brij 35 and 1.5mM EDTA, pH value is that it is thorough clearly that this growth hormone solution can keep when contained Tris HCl concentration was greater than 200mM in 7.4 the diluent.Shown in solution 6-8 in the table 4, remove and to contain 0.15% Brij 35 and 200mMTris HCl, PH is outside 7.2, also must have concentration greater than the EDTA of 1.5mM can make growth hormone solution keep 6 days thorough clearly.Solution 9-14 shows in the table 4, when diluent contains 0.15%Brij 35,2mM EDTA and or 1%, 5%, 10% sucrose solution, or 0.2M, 0.1M, the 0.0M sodium chloride solution, when the final pH value of sample is 7.8-7.9, the solution of growth hormone can keep 6 days thorough clearly.
Claims (32)
1. an energy still can keep thorough clearly growth hormone solution in one period long period He under the mechanical oscillation, be dissolved in diluent by a kind of cryodesiccated somatotropin composition and form, this cryodesiccated somatotropin composition comprises that the preceding pH value of every 1 part of growth hormone of 2-8 part arginine hydrochloride (weight portion) lyophilization is 7.2-8.5; This diluent then contains EDTA, and a kind of non-ionic surface activator is if between the pH value 7.2-7.8 of the preceding somatotropin composition of lyophilization, also contain a kind of buffer agent.
2. the pH value before the growth hormone solution of claim 1, somatotropin composition lyophilization wherein was more than or equal to 7.8 o'clock, and diluent also can contain a kind of non-buffer medium compatible with human or animal's receptor.
3. the pH value before the growth hormone solution of claim 1, somatotropin composition lyophilization wherein was more than or equal to 7.8 o'clock, and diluent also can contain a kind of buffer agent compatible with human or animal's receptor.
4. the growth hormone solution of claim 1,2 or 3, growth hormone wherein can be the growth hormone of natural or synthetic cattle, pig, people, bird, sheep or horse or the part analog of its biologically active.
5. the growth hormone solution of claim 4, growth hormone wherein is pig growth hormone (PST).
6. the growth hormone of claim 5, wherein pig growth hormone is δ-7 pig growth hormone.
7. the growth hormone of claim 4, wherein growth hormone is the growth hormone of melts combine.
8. the growth hormone solution of claim 1,2 or 3, wherein the non-ionic surface activator is polyoxyethylene-23-Laurel ether (background of cloth agent Brij35), Tween 80 or polyoxyethylene-20-cetyl ether (background of cloth agent Brij58), concentration is 0.08-2.0%.
9. the growth hormone solution of claim 8, wherein the non-ionic surface activator is brij35, its concentration is more than or equal to 0.08%.
10. the growth hormone solution of claim 9, when wherein the non-ionic surface activator was Brij35, concentration was 0.1-0.2%.
11. the growth hormone solution of claim 8, when wherein the non-ionic surface activator was Tween 80 or Brij35, its concentration was 0.08%-1.0%.
12. the growth hormone solution of claim 1 or 2, wherein buffer agent is the neutral pH buffer agent, and concentration is 0.2M-0.5M.
13. the growth hormone solution of claim 12, wherein buffer agent is Tris HCl or phosphate buffer.
14. the growth hormone solution of claim 13, wherein buffer agent is Tris HCl, and its concentration is 0.2M-0.3M.
15. the growth hormone solution of claim 2, wherein the concentration of non-buffer composition is 0.1M-0.5M.
16. the growth hormone solution of claim 15, wherein non-buffer composition is sucrose, trehalose or sodium chloride.
17. claim 1,2,3 growth hormone solution, if pH value less than 7.6, wherein the concentration of EDTA is more than or equal to 1.5mM; If the solution pH value is more than or equal to 7.6, wherein the concentration of EDTA is then more than or equal to 1.0mM.
Keep thorough clearly 18. be formulated under the mechanical oscillation, can not become the instrument of muddy growth hormone solution in one period persistent period, it comprises:
Adorn the bottle of the cryodesiccated somatotropin composition of being made by every 1 part of growth hormone of 2-8 part arginine hydrochloride (weight ratio) in first, wherein the pH value scope before this somatotropin composition lyophilization is 7.2-8.5; With
The bottle of dress diluent in second, diluent contains EDTA, a kind of non-ionic surface activator, if the pH value of somatotropin composition is between 7.2-7.8 before the lyophilization, diluent also contains a kind of buffer agent, and wherein keeping thorough clearly and can not become muddy solution when standing mechanical oscillation in one period persistent period is to be dissolved in second inclusions in the bottle by the inclusions with first bottle forming.
19. the instrument in the claim 18 is if wherein the pH value of the preceding somatotropin composition of lyophilization also contained a kind of non-buffer medium of biocompatible more than or equal to 7.8 o'clock in the diluent.
20. the instrument in the claim 18 is if wherein the pH value of the preceding somatotropin composition of lyophilization also contained a kind of buffer agent compatible with receptor more than or equal to 7.8 o'clock in the diluent.
21. the instrument of claim 18,19 or 20, wherein every milliliter of cryodesiccated somatotropin composition of diluent dissolving 0.5mg-40mg.
22. the instrument of claim 18,19 or 20, growth hormone wherein are growth hormone or its part with growth activity or the analog of natural or synthetic cattle, pig, people, bird, sheep or horse.
23. the instrument of claim 18,19 or 20, surfactant wherein are polyoxyethylene-23-Laurel ether (Brij35), Tween 80 or polyoxyethylene-20 its concentration of cetyl ether (Brij58) are 0.08%-2.0%.
24. the instrument of claim 18 or 20, buffer agent wherein are Tris HCl, phosphate, and concentration is 0.2M-0.5M.
25. the instrument of claim 19, non-buffer composition wherein is sucrose, trehalose or sodium chloride, and concentration is 0.1M-0.5M.
26. be formulated under the mechanical vibration method that keeps clear thorough and longer a period of time still to keep thorough clearly growth hormone solution, may further comprise the steps:
(1) prepare cryodesiccated somatotropin composition: the dissolving arginine hydrochloride is in growth hormone, ratio is that 1 part of growth hormone is than 2-8 part arginine hydrochloride, regulate pH value to the 7.2-8.5 scope, remove insoluble matter, carry out lyophilization to generating solution then with filtration and centrifuging;
(2) the cryodesiccated somatotropin composition that will generate is dissolved in the diluent that contains EDTA and a kind of non-ionic surface activator, if the pH value of somatotropin composition is between 7.2-7.8 before the lyophilization, this diluent also contains a kind of buffer agent.
27. the method for claim 26, wherein growth hormone synthetic pH value is more than or equal to 7.8 before the lyophilization, and diluent also contains a kind of non-buffer medium of biocompatible, and its concentration range is 0.1M-0.5M.
28. the method for claim 26, wherein this pH value of somatotropin composition was more than or equal to 7.8 o'clock before the lyophilization, and diluent also contains a kind of neutrality and receptor compatible buffers, and its concentration range is 0.2M-0.5M.
29. the method for claim 26 or 28, buffer agent wherein are Tris HCl or phosphate buffer.
30. the method for claim 27, non-buffer composition wherein is sucrose, trehalose or sodium chloride.
31. the method for claim 26,27 or 28, wherein growth hormone is meant the growth hormone of natural or synthetic cattle, pig, people, bird, sheep or horse or the part or the analog of its biologically active.
32. the method for claim 26,27 or 28, wherein the non-ionic surface activator is polyoxyethylene-23 a Laurel ether (Brij35), Tween 80 or polyoxyethylene-20 cetyl ether (Brij58), and concentration is 0.08%-2.0%.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US86476892A | 1992-04-07 | 1992-04-07 | |
| US864,768 | 1992-04-07 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1078910A CN1078910A (en) | 1993-12-01 |
| CN1089249C true CN1089249C (en) | 2002-08-21 |
Family
ID=25344034
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN93104030A Expired - Fee Related CN1089249C (en) | 1992-04-07 | 1993-04-06 | Biocompatible somatotropin solutions |
Country Status (6)
| Country | Link |
|---|---|
| EP (1) | EP0639984A1 (en) |
| JP (1) | JPH08502021A (en) |
| CN (1) | CN1089249C (en) |
| AU (1) | AU4278393A (en) |
| CA (1) | CA2133737A1 (en) |
| WO (1) | WO1993019773A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103751769A (en) * | 2013-12-25 | 2014-04-30 | 楼秀余 | Lyophilized recombinant human growth hormone capsule and preparation method thereof |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5981485A (en) | 1997-07-14 | 1999-11-09 | Genentech, Inc. | Human growth hormone aqueous formulation |
| US5898030A (en) * | 1994-06-17 | 1999-04-27 | Applied Research Systems Ars Holding N.V | hGH containing pharmaceutical compositions |
| KR100236393B1 (en) | 1996-02-02 | 1999-12-15 | 나까니시 히로유끼 | A pharmaceutical preparation containing a human growth hormone |
| KR100890679B1 (en) | 2000-02-24 | 2009-03-26 | 일라이 릴리 앤드 캄파니 | Non-aqueous injectable formulations for extended release of somatotropin |
| AU2001271491A1 (en) | 2000-06-26 | 2002-01-08 | Monsanto Technology Llc | Non-aqueous surfactant-containing formulations for extended release of somatotropin |
| US6664234B1 (en) | 2000-06-30 | 2003-12-16 | Monsanto Technology Llc | Non-aqueous injectable formulation preparation with pH adjusted for extended release of somatotropin |
| MX349301B (en) | 2009-12-21 | 2017-07-21 | Ambrx Inc | Modified bovine somatotropin polypeptides and their uses. |
| BR112012015597A2 (en) | 2009-12-21 | 2017-01-31 | Ambrx Inc | modified porcine somatotropin peptides and their uses |
| ES2715028T3 (en) | 2012-06-05 | 2019-05-31 | Neuroderm Ltd | Compositions comprising apomorphine and organic acids and their uses |
| ITBO20120368A1 (en) | 2012-07-06 | 2014-01-07 | Alfa Wassermann Spa | COMPOSITIONS INCLUDING RIFAXIMINA AND AMINO ACIDS, RIFAXIMINE CRYSTALS DERIVING FROM SUCH COMPOSITIONS AND THEIR USE. |
| GB201602880D0 (en) | 2016-02-18 | 2016-04-06 | Ge Healthcare Uk Ltd | Method and composition for biomolecule stabilization |
| CN112618482A (en) * | 2019-09-24 | 2021-04-09 | 江苏恒瑞医药股份有限公司 | Novel protein formulations |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0303746A1 (en) * | 1987-08-21 | 1989-02-22 | Mallinckrodt Group Inc. | Stabilization of growth promoting hormones |
| WO1990002560A1 (en) * | 1988-09-12 | 1990-03-22 | Pitman-Moore, Inc. | Swine growth promotion |
| WO1990014100A2 (en) * | 1989-05-15 | 1990-11-29 | Pitman-Moore, Inc. | Somatotropin for increasing fertility in animals |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3325223A1 (en) * | 1983-07-13 | 1985-01-24 | Hoechst Ag, 6230 Frankfurt | AGAINST AGAINST DENATURATION, AQUEOUS PROTEIN SOLUTIONS, METHODS FOR THEIR PRODUCTION AND THEIR USE |
| US4774091A (en) * | 1983-10-14 | 1988-09-27 | Sumitomo Pharmaceuticals Company, Ltd. | Long-term sustained-release preparation |
-
1993
- 1993-04-05 EP EP93912120A patent/EP0639984A1/en not_active Withdrawn
- 1993-04-05 JP JP5517756A patent/JPH08502021A/en active Pending
- 1993-04-05 WO PCT/US1993/003182 patent/WO1993019773A1/en not_active Application Discontinuation
- 1993-04-05 CA CA002133737A patent/CA2133737A1/en not_active Abandoned
- 1993-04-05 AU AU42783/93A patent/AU4278393A/en not_active Abandoned
- 1993-04-06 CN CN93104030A patent/CN1089249C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0303746A1 (en) * | 1987-08-21 | 1989-02-22 | Mallinckrodt Group Inc. | Stabilization of growth promoting hormones |
| WO1990002560A1 (en) * | 1988-09-12 | 1990-03-22 | Pitman-Moore, Inc. | Swine growth promotion |
| WO1990014100A2 (en) * | 1989-05-15 | 1990-11-29 | Pitman-Moore, Inc. | Somatotropin for increasing fertility in animals |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103751769A (en) * | 2013-12-25 | 2014-04-30 | 楼秀余 | Lyophilized recombinant human growth hormone capsule and preparation method thereof |
| CN103751769B (en) * | 2013-12-25 | 2015-08-19 | 楼秀余 | A kind of lyophilizing recombinant human somatropin capsule and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH08502021A (en) | 1996-03-05 |
| CN1078910A (en) | 1993-12-01 |
| EP0639984A1 (en) | 1995-03-01 |
| AU4278393A (en) | 1993-11-08 |
| CA2133737A1 (en) | 1993-10-14 |
| WO1993019773A1 (en) | 1993-10-14 |
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