CN104606679A - Production method for composite lactose for injection - Google Patents

Abstract

The invention relates to a production technology for composite lactose for injection. According to the technology, ultrafiltration, nanofiltration and recrystallization are combined to remove residual glucose, fructose, galactose, amino acid and protein in lactose; and the prepared lactose conforms to the requirements for injection, and can be applied to an injection product. The production technology provided by the invention is simple, feasible, relatively high in yield, and suitable for industrial production.

Description

A kind of production method of injection compound lactose

Technical field

The invention belongs to field of medicaments, be specifically related to a kind of production method of injection compound lactose.

Background technology

Lactose is a kind of disaccharidase, is made up of, under field conditions (factors) a part glucose and a part galactose, be present in mammiferous milk, in commercial production, often extract from milk surum, containing a small amount of glucose, galactose, fructose, protein, aminoacid in lactose crude product.At field of medicaments, lactose has important application, as lyophilized preparation support, it is compared with other conventional lyophilized preparation (such as: glucose, mannitol etc.) maximum difference, and the latter is monosaccharide, and molecular mass is relatively low, glass transition temperature during lyophilizing is also lower, and freeze-drying process is also a crystallization process, some is compared to the lyophilized preparation of " fragility ", this crystallization process has extremely strong destruction; The lactose that glass transition temperature is higher is in addition also nodeless mesh generation in freeze-drying process; but with unformed existence; destruction do not had to main constituent or some special preparation, the role that therefore in this kind of preparation, lactose all plays usually " freeze drying protectant ".

Due to numerous good characteristic of lactose, make it in lyophilized formulations field, especially there is in the application of Novel Drug Delivery Systems (nano-emulsion, liposome etc.) irreplaceable effect.But because source is more special, its lactose produced through traditional purifying process, owing to there is many impurity, very easily cause a series of untoward reaction such as such as urticaria, cough, heating, shock when drug administration by injection, this makes the application of lactose in injection product of function admirable originally be greatly limited.

The process for purification of lactose disclosed in Chinese patent application 201210437448.0, the method is for the production of the lactose of injection, but when not removing fructose, glucose, galactose, amino acid and protein in lactose crude product, adopt the high-temperature digestion of 50 ~ 70 DEG C, easy generation Maillard reaction, the Fructoamino-acid particularly generated forms infringement to human body.

Current purifying process does not still reach the requirement of production injection stage lactose, therefore in the urgent need to the technique of new purification lactose, to prepare a kind of safe and reliable lactose for injection.

Summary of the invention

Not high in order to solve current pharmaceutical lactose purity, there is residual protein, do not reach this problem of injection requirement, and frozen-dried protective usefulness is still not enough, the invention provides a kind of purification process of lactose, the lactose after purification can be used for injection product.

The method purification lactose that the present invention adopts nanofiltration and recrystallization to combine,

Compared with the process for purification of lactose described in patent application 201210437448.0, the filter method that the present invention adopts ultrafiltration and nanofiltration to combine, remove protein, virus and antibacterial, molecular cut off 200 ~ 1000 material, obtain the lactose that purity is higher, then adopt high-temperature digestion, then remove protein further by the method for alcohol water crystallization, and the whole content of ethanol controls, between 70 ~ 75%, further to sterilize while crystallisation by cooling to system.

Technical scheme of the present invention is as follows:

A production method for medicinal compound lactose, the production craft step of described medicinal compound lactose is as follows:

1. get lactose crude product appropriate, add the water of 2.4 ~ 4 times of weight, be heated with stirring to 30 ~ 40 DEG C, make lactose solution;

2. lactose is filtered through the ultrafiltration apparatus that combined closure system is 3000MW;

3. filtrate is filtered through nanofiltration device;

4. by residue collection, add the purified water of 1.3 ~ 2.5 times of weight, be heated with stirring to 50 ~ 70 DEG C, make lactose solution;

5. by the lactose solution in step 4, add the ethanol of 3 ~ 3.8 times of volumes, stir and be cooled to 10 DEG C, crystallize out, this process keeps 0.1 ~ 5 hour;

6. the solution centrifugal will crystal being had to separate out, collects crystal;

7. by the crystal of step (6) gained and viscous solution Homogeneous phase mixing;

8. spraying dry, to obtain final product.

Lactose crude product described in step 1 is commercially available common lactose or pharmaceutical lactose;

Described viscous solution is hydroxypropyl emthylcellulose aqueous solution.

The wherein lactose of compound lactose by 95%-99% and the hydroxypropyl emthylcellulose of 1%-5%.

The present invention, by adopting the method for ultrafiltration, nanofiltration and alcohol water recrystallization, reaches more than 99.99% to the clearance rate of protein, reaches more than 99%, facilitate the high-temperature digestion of subsequent step to fructose, glucose, amino acid whose clearance rate.The lactose residual protein content that the present invention obtains significantly reduces; can reduce in Clinical practice because of untoward reaction that heterologous protein causes; and glycosyl aminoacid can not be produced in high-temperature digestion process; especially Fructoamino-acid; decrease the harm to human body; solve the problem of a series of untoward reaction of normal oral rank lactose for existing during injection, and greatly improve frozen-dried protective usefulness.Can be used in injection, for the exploitation of pharmaceutical industry new administration injection dosage form provides a kind of excellent adjuvant.Purifying process simple possible provided by the invention, yield is high, and cost is lower, environmental friendliness, is suitable for the large production of industry.

Detailed description of the invention

Come by the following examples to be described further content of the present invention.

Embodiment 1:

1. take lactose crude product 100g, add 400ml purified water, be heated with stirring to 30 DEG C, make it dissolve formation lactose solution completely;

2. the ultrafiltration post being 3000MW by lactose crude product solution molecular cut off filters;

3. the lactose solution after ultrafiltration is carried out nanofiltration;

4. the lactose leached is added in 150ml purified water, be heated with stirring to 70 DEG C, make it be dissolved into lactose solution completely;

5. add 450ml ethanol by the lactose solution in step 4, stir and be cooled to 10 DEG C, crystallize out, this process keeps 0.5 hour;

6. the solution having crystal to separate out is carried out centrifugal through bag-filter type centrifugal machine, collect crystal;

7, get 99 grams of crystal and 100ml contain the Gonak stirring and evenly mixing of 1% after spraying dry and get final product.

Embodiment 2:

1. take lactose crude product 20Kg, add 62L purified water, be heated with stirring to 40 DEG C, make it dissolve formation lactose solution completely;

2. the ultrafiltration post being 3000MW by lactose crude product solution molecular cut off filters;

3. the lactose solution after ultrafiltration is carried out nanofiltration;

4. the lactose leached is added in 30L purified water, be heated with stirring to 70 DEG C, make it be dissolved into lactose solution completely;

5. add 100L ethanol by the lactose solution in step 4, stir and be cooled to 10 DEG C, crystallize out, this process keeps 1 hour;

6. the solution having crystal to separate out is carried out centrifugal through bag-filter type centrifugal machine, collect crystal;

7. get 97 grams of crystal and 100ml contain the Gonak stirring and evenly mixing of 3% after spraying dry and get final product.

Embodiment 3:

1. take lactose crude product 20Kg, add 62L purified water, be heated with stirring to 35 DEG C, make it dissolve formation lactose solution completely;

2. the ultrafiltration post being 3000MW by lactose crude product solution molecular cut off filters;

3. the lactose solution after ultrafiltration is carried out nanofiltration;

4. the lactose leached is added in 32L purified water, be heated with stirring to 70 DEG C, make it be dissolved into lactose solution completely;

5. add 100L ethanol by the lactose solution in step 4, stir and be cooled to 10 DEG C, crystallize out, this process keeps 1 hour;

6. the solution having crystal to separate out is carried out centrifugal through bag-filter type centrifugal machine, collect crystal;

7, get 98 grams of crystal and 100ml contain the Gonak stirring and evenly mixing of 2% after spraying dry and get final product.

Embodiment 4:

1. take lactose crude product 25Kg, add 80L purified water, be heated with stirring to 40 DEG C, make it dissolve formation lactose solution completely;

2. the ultrafiltration post being 3000MW by lactose crude product solution molecular cut off filters;

3. the lactose solution after ultrafiltration is carried out nanofiltration;

4. the lactose leached is added in 60L purified water, be heated with stirring to 50 DEG C, make it be dissolved into lactose solution completely;

5. add 180L ethanol by the lactose solution in step 4, stir and be cooled to 10 DEG C, crystallize out, this process keeps 1 hour;

6. the solution having crystal to separate out is carried out centrifugal through bag-filter type centrifugal machine, collect crystal;

7, get 95 grams of crystal and 100ml contain the Gonak stirring and evenly mixing of 5% after spraying dry and get final product.

Experimental example properties of product detect

According to lactose (injection) standard in Chinese Pharmacopoeia 2015 version two (exposure draft), detect the part important indicator of embodiment 1-4 product, result is as follows:

As can be seen from the above results, the lactose that the present invention produces can reach the injection standard of Chinese Pharmacopoeia, and part index number is more better than set standard value.

Comparative example: according to the preparation of embodiment 1 method, difference is that water-soluble Direct spraying is dry after preparing crystal.

Survival experiment:

For lactobacillus rhamnosus, cultivate being inoculated in MRS liquid culture medium by 3% inoculum concentration after actication of culture at 37 DEG C, every 3 h sampling and measuring viable counts, take incubation time as abscissa, viable count draws growth curve, to understand the growth cycle of this bacterium for vertical coordinate.RS culture medium: peptone 1%, Carnis Bovis seu Bubali cream 1%, glucose 3%, sodium acetate 0.5%, yeast extract 0.5%, dibasic ammonium citrate 0.2%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.02%, Tween800.1% (pH6.5,121 DEG C, 30 min sterilizings).The activation of strain, takes out an inclined-plane and access sterilized liquid MRS culture medium with Inoculating needle picking 2 ring lactobacillus casei under sterile working's state, cultivate 24 ~ 48 h for 37 DEG C and activate, activate 2 times with for subsequent use according to above-mentioned same steps from refrigerator.Amplification culture, by the seed liquor that activated with in the inoculum concentration access MRS liquid triangular flask culture medium of 3%, cultivates 18 h for 37 DEG C.The collection of thalline, puts into centrifuge with 13 min centrifugal under 4500 r/min, abandoning supernatant after the fermentation liquid after cultivation is weighed up quality.Put into sterilized 20 mL small beakers after bacterium mud and protective agent aseptically being mixed, in-80 DEG C of refrigerators, carry out pre-freeze, put into freeze dryer after the preservative film on small beaker being pricked hole after 2 h and carry out lyophilizing.

Count plate: after adopting methylene blue counting method diluted sample certain multiple to make bacteria suspension, move pipe with 5 μ L micro quantitative determinations and inhale

Getting bacteria suspension puts on clean slide, add the 0.1% methylene blue dye liquor of 5 μ L, covered, attention can not have bubble, also can not bacterium hydrorrhea be gone out beyond coverslip, methylene blue is soaked sheet put microscope ocular 16 ×, objective lens 40 × lower microscopy, select 10 ~ 20 visuals field of leaching sheet 4 angles and central authorities by distribution and calculate viable bacteria body number, obtain the lactic acid viable count meansigma methods A in each visual field, visual field area is B, and coverslip area is C, extension rate is D, then the viable count in every milliliter or every gram of sample is: C/B × A × (1000/5) × D.

Protective agent is respectively the compound lactose of embodiment 1, the lactose of comparative example, maltose and trehalose, and concentration is 5%, and result of the test is as follows:

Freeze drying protectant	Embodiment 1	Comparative example	Maltose	Trehalose
					Survival rate	74.3％	25.1％	44.1％	55.2％

Claims (5)

1. a production method for injection compound lactose, comprises the following steps:

(1) get lactose crude product, add the purified water of 2.4 ~ 4 times of weight, be heated with stirring to 30 ~ 40 DEG C, make lactose solution;

(2) lactose solution is carried out ultrafiltration through the ultrafiltration apparatus that molecular cut off is 3000MW;

(3) lactose solution after ultrafiltration is carried out nanofiltration through nanofiltration device;

(4) get the lactose filtering residue after retaining, add the purified water of 1.3 ~ 2.5 times of weight, be heated with stirring to 50 ~ 70 DEG C, make lactose solution;

(5) by the lactose solution in step (4), add the ethanol of 2.3 ~ 3 times of volumes, stir and be cooled to 10 DEG C, crystallize out, this process lasts 0.1 ~ 5 hour;

(6) solution centrifugal will crystal being had to separate out, collects crystal;

(7) by the crystal of step (6) gained and viscous solution Homogeneous phase mixing;

(8) spraying dry, to obtain final product.

2. the production method of a kind of injection compound lactose as claimed in claim 1, is characterized in that the pressure reduction that the ultrafiltration in described step (2) adopts is 0.03 ~ 0.6MPa.

3. the production method of a kind of injection compound lactose as claimed in claim 1, is characterized in that the pressure reduction of the nanofiltration in described step (3) is 0.5 ~ 1.0MPa.

4. the production method of a kind of injection lactose as claimed in claim 1, it is characterized in that, described viscous solution is hydroxypropyl emthylcellulose aqueous solution, wherein the lactose of compound lactose by 95%-99% and the hydroxypropyl emthylcellulose of 1%-5%.

5. produced the injection compound lactose obtained by the method described in any one of claim 1-4 for one kind.