CN101822695A

CN101822695A - Complex bone peptide injection and preparation method of complex bone peptide for injection

Info

Publication number: CN101822695A
Application number: CN201010137762A
Authority: CN
Inventors: 石海
Original assignee: Individual
Current assignee: Individual
Priority date: 2010-04-02
Filing date: 2010-04-02
Publication date: 2010-09-08

Abstract

The invention discloses a complex bone peptide injection and complex bone peptide for injection, which are sterile water solution prepared by extracting limb bones of healthy pigs or fetal bovines and scorpions, and a freeze-drying agent prepared by adding a proper amount of auxiliary materials into the sterile water solution and performing freeze-drying on the mixture. The complex bone peptide injection comprises the main components of organic calcium, phosphorus, inorganic calcium, inorganic salts, trace elements, amino acid and the like. The complex bone peptide injection is used for treating rheumatism, rheumatoid arthritis, hyperosteogeny and bone fracture. The complex bone peptide injection and the complex bone peptide for injection belong to the technical field of medicaments. The invention introduces a preparation method, which comprises the process stops of: performing bone extraction, degreasing, concentration, hydrolysis, acid deposition, alkali deposition and ultrafiltration to obtain stock solution of a bone extract; performing scorpion cleaning, mincing, ethanol extraction, ethanol recycling and ultrafiltration to obtain stock solution of a scorpion extract; and mixing the bone extract with the scorpion extract in proportion, and performing ultrafiltration concentration, encapsulation and sterilization to obtain the complex bone peptide injection. The concentrated solution is added with a proper amount of excipient, and then the mixture is sterilized and freeze-dried to obtain the complex bone peptide for injection. The whole process is simple, convenient and complete to ensure that a product accord with a national standard, viruses are inactivated, and simultaneously the sterilization F0 of the injection is more than 8, and the sterility assurance level is high.

Description

The preparation method of a kind of complex bone peptide for injection and complex bone peptide injection

Technical field:

The invention belongs to medical technical field

Background technology

Complex bone peptide for injection and complex bone peptide injection are the sterile water solutions of being made by the pig of health or fetal bovine limb bone and Scorpio and add an amount of lyophilized preparation that the material lyophilizing is made of composing.Main component is organic calcium, phosphorus, inorganic calcium, inorganic salt, trace element, aminoacid etc.Be used for the treatment of rheumatism, rheumatoid arthritis, hyperosteogeny, fracture.

This kind is China's the first, and its technical process document has many places reports, but by our practical operation, all can't make qualified products, the present invention describes a kind of manufacture method in detail, and simple, suitable, safety can reach the specific requirement of the national drug standards fully.

Summary of the invention is

The extraction of a, bone, degrease, concentrated: get healthy fresh pig or fetal bovine limb bone, clean, smash and weigh.The per kilogram bone adds water 1-3kg, in 1.1-1.3kg/cm ²Hot pressing 1-3 hour, filter with double gauze, the bone slag adds the water commensurability with green bone again, and the same operation hot pressing 1-2 hour is filtered, and merges filtrate twice.In the jacketed pan internal cooling to room temperature, be positioned in-10 to-30 ℃ of freezers, to be cooled changing over to 0～5 ℃ left standstill 12～24 hours in the cold house, skim upper strata fat, heat to make and freeze the shape thing and melt into liquid, can heat and be concentrated into the per kilogram extract and contain and be equivalent to 0.5 kilogram of bone.

B, hydrolysis: the said extracted thing is transferred about pH2.5-3.5, press bone heavy 1～3% add pepsin, in 38 ℃ to 42 ℃ hydrolysis 3-5 hour.

C, acid precipitation: above-mentioned liquid in 100 ℃ of heating about 30 minutes, is continued acid hydrolysis, cooling sucking-off supernatant.

D, alkali deposited: above-mentioned supernatant is transferred pH8-9, is heated to boiling, continues about 30 minutes, continues the basic hydrolysis cooling, sucts clearly.

E, ultrafiltration: above-mentioned supernatant is transferred pH to 7.0-7.5, is filtered to clearly, crosses 10,000 molecular weight ultrafiltration 1-3 time, promptly gets bone extract solution.

The cleaning of f, Scorpio, rub: an amount of Scorpio of weighing, wash secondary with pure water, shred with shears.

The ethanol extraction of g, Scorpio: two times (W/V) heavy by scorpion adds 95% ethanol, and stir and extracted about 5 hours, the static after-filtration that spends the night, secondary merges twice ethanol extract repeatedly.

H, reclaim ethanol: the ethanol extraction vacuum concentration adds distilled water to full volumetric about half, steams half volume again, for several times to there being the ethanol flavor, adds water to the twice of Scorpio weight repeatedly, concentrated solution.

I, layered filtration: the concentrated solution static layering is spent the night, tell lower aqueous solution then, the upper strata fat deposit discards, and the aqueous solution of extraction merges.

J, ultrafiltration: 10,000 molecular weight ultrafiltration are 1-3 time after the aqueous solution coarse filtration, the weighing volume.

K, bone extracting solution mix with the Scorpio extracting solution

Extract by containing the 0.015g Scorpio among the every 1m1 of final volume mixes Scorpio extract with bone extract.

1, ultrafiltration concentrates

Above-mentioned mixed liquor is crossed 10,000 molecular weight ultrafiltration posts 1-3 time, and ultrafiltrate is concentrated, and extremely polypeptide and nitrogen content are qualified among every ml, contain Scorpio extract simultaneously and are equivalent to the 0.015g Scorpio.

M, preparation

Complex bone peptide solution dilution or be concentrated into the desired concn scope, also can add an amount of sodium chloride and transfer to etc. and to ooze, regulate pH value in the sign scope, aseptic filtration, embedding, also can cross 10,000 molecular weight ultrafilters once after, aseptic filtration, embedding, in 100 ℃ of pressure sterilizings 30 minutes or 115 ℃ 30 minutes, 121 ℃ sterilizations in 12 minutes, 15 minutes, check, lamp inspection, packing promptly gets complex bone peptide for injection.After complex bone peptide solubilize thoroughly, add an amount of mannitol or dextran as excipient (being generally the 10%-15% of solution amount), cross 0.22 μ m microporous filter membrane, fill, lyophilization promptly gets complex bone peptide injection.

It is characterized in that:

A, manufacture process are easy, and the suitability is strong.

B, soda acid repeatedly is heat sink, has guaranteed that the preparation visible foreign matters is qualified.

C, 121 ℃ of hot pressing in 12-30 minute of this process using have guaranteed the inactivation of virus condition, and are also qualified favourable to the visible foreign matters of preparation.

D, injection adopt 121 ℃ of sterilizations in 12-30 minute, have guaranteed F ₀Value is greater than 8, aseptic safeguard level height.

Concrete embodiment

Get healthy fresh pig bones of limbs 75kg, clean, smash and weigh.Add water 75kg, in 1.2kg/cm ²Hot pressing 2 hours is filtered with double gauze, and the bone slag adds the water 75kg commensurability with green bone again, and the same operation hot pressing 1-2 hour is filtered, and merges filtrate twice., be positioned in-10 to-30 ℃ of freezers to room temperature in the jacketed pan internal cooling, to be cooled changing over to 0～5 ℃ left standstill 12～24 hours in the cold house, skim upper strata fat, heats to make to freeze the shape thing and melt into liquid.The said extracted thing is transferred about pH2.5-3.5, pressed 2% heavy adding pepsin of bone, in 38 ℃ to 42 ℃ hydrolysis 4 hours.Above-mentioned liquid in 100 ℃ of heating 30 minutes, is continued acid hydrolysis, cooling sucking-off supernatant.Above-mentioned supernatant is transferred about pH8.5, is heated to boiling, continues 30 minutes, continues the basic hydrolysis cooling, sucts clearly.Above-mentioned supernatant is transferred pH to 7.4, is filtered to clearly, crosses 10,000 molecular weight ultrafiltration secondaries, promptly gets the about 150kg of bone extract solution, and recording peptide concentration is 8.5mg/ml.

Weighing 300g Scorpio is washed secondary with pure water, shreds with shears. and two times (W/V) heavy by scorpion adds 95% ethanol, and stir and extracted 5 hours, the static after-filtration that spends the night, secondary merges twice ethanol extract repeatedly.The ethanol extraction vacuum concentration adds distilled water to full volumetric about half, steams half volume again, for several times to there being the ethanol flavor, adds water to the twice of Scorpio weight repeatedly, concentrated solution.The concentrated solution static layering is spent the night, tell lower aqueous solution then, the upper strata fat deposit discards, and the aqueous solution of extraction merges.After the aqueous solution coarse filtration 10,000 molecular weight ultrafiltration once, the weighing volume.Get the about 600ml of scorpion extract.

Get the extract that bone extract solution 8000ml adds the above-mentioned 60g of being equivalent to Scorpio, cross 10,000 molecular weight ultrafilter secondaries, above-mentioned mixed liquor is concentrated into 4000ml.

Above-mentioned solution is regulated pH value in the sign scope, and measures peptide concentration and nitrogen concentration, the aseptic filtration embedding of qualified back, and in 121 ℃ of pressure sterilizings 15 minutes, lamp inspection, packing promptly got complex bone peptide for injection.Above-mentioned complex bone peptide solution is added 12% mannitol except that thermal source, cross 0.22 μ m microporous filter membrane, fill, lyophilization promptly gets complex bone peptide injection.

Claims

1. extract on the spissated basis of degrease in bone hot pressing, with HCL adjust pH 2.5-3.5, press the heavy 1-3% of bone and add pepsin, in 38 to 42 ℃ hydrolysis 3-5 hour, in 100 ℃ of heating 30 minutes, supernatant is drawn in cooling, and supernatant is with NaOH adjust pH 8-9, in 100 ℃ of heating 30 minutes, supernatant is drawn in cooling, with HCL adjust pH 7.0-7.5, cross 10,000 molecular weight ultrafilters 1-3 time, get bone extract solution.

2. take by weighing an amount of Scorpio, use the pure water washed twice, shred, (W/V) 95% ethanol that adds twice stirs and extracted about 5 hours, static spending the night, repeatedly twice, merge extracted twice liquid, vacuum concentration is to there being the ethanol flavor, thin up, the static layering of spending the night gets lower layer of water liquid, clarification filtration, after 10,000 molecular weight ultrafilters 1-3 time, the scorpion extract.

3. contain the extract that is equivalent to the 0.015g Scorpio by the every ml of final volume, Scorpio extract is mixed with bone extract, cross 10,000 molecular weight ultrafilters 1-3 time, it is qualified that ultrafiltrate heating is concentrated among every ml polypeptide and nitrogen content, contain Scorpio extract simultaneously and be equivalent to the 0.015g Scorpio, with above-mentioned concentrated solution adjust pH in the sign scope, aseptic filtration, embedding also can be crossed aseptic filtration embedding again behind the 10000 molecular weight ultrafilters, in 121 ℃ of pressure sterilizings 15 minutes, check, lamp inspection promptly gets complex bone peptide for injection, it is excipient (being generally the 10-15% of solution amount) that above-mentioned concentrated solution adds an amount of mannitol or dextran, aseptic filtration, fill, lyophilization, sealing by fusing promptly gets complex bone peptide injection.