CN101549146B - Romurtide injection and preparation method thereof - Google Patents

Abstract

The invention provides a romurtide injection which comprises the effective dose of romurtide, solvent, pH regulator and buffer solution which function as active components. The preparation method of the romurtide injection comprises the following steps: the prescribed romurtide is put in a proper vessel, the prescribed solvent is added in the vessel for dissolution, the buffer solution is added until the total amount required is achieved, the mixture is processed by water bath and stirred for solution, the pyrogen is removed through an ultrafilter, the mixture is filtered by a microporous filter membrane and sterilized, and then the subpackaging and the top sealing are performed.

Description

Romurtide injection and preparation method thereof

Invention field

The present invention relates to a kind of leukopenic Romurtide injection that causes for chemotherapy or radiotherapy and preparation method thereof.

Technical background

Malignant tumor has become one of serious disease that threatens human health and life.According to statistics, annual new cancer diagnosis patient more than 1,000 ten thousand people in the whole world, patient's sum 4,000 ten thousand people, dead 7,100,000 people are with infection and cardiovascular and cerebrovascular disease row fatality rate front three.China's malignant tumor fall ill every year on average 2000000 people, dead 1,600,000 people, account for the whole nation every year on average total death toll 18.63%, occupy first of all kinds of causes of the death in the urban area.Antineoplastic agent and immunomodulator are positioned at the 5th of domestic hospitals medication sales volume (accounting for 13.44%).Chemotherapy and radiation is the important means of anticancer therapy and has obtained significant curative effect, but because of its toxic and side effects large, as suppress medulla hematopoietic system and cause that leukocyte and thrombocytopenia are with infecting and hemorrhage danger, hindered its use, more patient makes immunologic function degression because of tumor and treatment thereof, causes the vicious cycle threat to life.

Romurtide belongs to muramyldipeptide para-immunity reinforcing agent, it is the immune active ingredient of mycobacteria cell wall skeleton (must be good such as born of the same parents), it can produce the hematopoietic regulation factor by the activated mononuclear macrophage system, have simultaneously leukocyte increasing and hematoblastic characteristics, be mainly used in clinically the Secondary cases bone marrow depression that anticancer therapy (chemotherapy or radiotherapy) produces, promote peripheral blood leucocyte and platelet count purpose to increase, can prevent patient infection, reduce chemotherapy dosage, shorten the interval course for the treatment of of chemotherapy, thereby effectively improve chemotherapy effect, compare with recombinant cytokine class medicine, have and promote simultaneously leukocyte and thrombocytopoiesis, recover immunologic function, the molecule minor structure is single, can synthesize in a large number, purity is high, steady quality is controlled easily, curative effect is low than high toxicity, the characteristics such as biological pollution is few.Romurtide promotes leukocyte and thrombocytopoiesis simultaneously, also is effectively for high-dose chemotherapy, if any document be exactly to have obtained good therapeutic effect with leukocyte and thrombocytopenia behind the romurtide treatment Study of High Dose Carboplatin Chemotherapy.When chemicotherapy begins, just generally use romurtide at Japanese tumour patient, become routine administration, make patient more stable during whole treatment, better tolerance.

Romurtide is at first synthetic by Japanese Shiba etc., and its chemical constitution is N2-(N-acetyl muramyl-L-alanyl-D-isoglutamine)-N 6-stearoyl-1B, is equivalent to pentapeptide, and molecular formula is C ₄₃H ₇₈N ₆O ₁₃, molecular weight is 887.12.Its stable chemical nature, can promote normally to reach the hemoposieis of bone marrow injury animal pattern (mice or monkey that Cavia porcellus, Canis familiaris L. or the mouse model that causes such as irradiation and cyclophosphamide or carboplatin cause), the leukocyte increasing number is neutrophilic granulocyte and mononuclear cell number especially, strengthen simultaneously that it is engulfed, the function such as chemotactic and secretion activity oxygen, the platelet count of Cavia porcellus, Canis familiaris L. and monkey is significantly increased; Neuromuscular, breathing and cardiovascular function to animal do not make significant difference, and do not have mutagenesis and teratogenesis yet.Japan the first drugmaker succeeds in developing, the listing that went through in 1991, and its preparation freeze-dried powder injection commodity are called Nopia, are mainly used in chemotherapy or Cancer Patients Undergoing Radiotherapy, promote leukocyte count and platelet count to return to rapidly normal level.

But because the romurtide physicochemical property is comparatively special, hardly dissolving in aqueous solution, and be very easy to change into gel rubber system after containing extremely micro-romurtide in the aqueous solution, therefore the dosage form of at present Japan's listing is the injection freeze-dried powder, but must use the in advance dissolving of special lyase during the said preparation injection, course of dissolution is very long, and the Romurtide injection outward appearance that obtains can not be clear and bright state fully, with certain opalescence, up to the present also there are not individual and mechanism that the problems referred to above are solved.Its clinical use of romurtide injection freeze-dried powder of at present Japan's listing need to be redissolved with special solvent, redissolution time length is made troubles to use, therefore opalescence is arranged after the dissolving and easily cause secondary pollution, it is significant to use clinically receptible suitable injection lyase to make stable, clear and bright Romurtide injection.The present invention now addresses these problems.

Summary of the invention

The invention provides a kind of Romurtide injection of suitable clinical practice.

Be to realize goal of the invention, the present invention by the following technical solutions:

Romurtide injection, this infusion pump contain the romurtide as active component of effective dose, and be consistent with the romurtide freeze-dried powder.

Described injection is aseptic parenteral solution.

Described injection also comprises solvent, pH adjusting agent and buffer solvent.

Described solvent is organic solvent.As: ethanol, glycerol, 1,2-PD, low molecular poly or their mixture.Preferred alcohol.We find that surprisingly romurtide is first with a small amount of ethanol or after containing the mixture dissolving romurtide of ethanol in experimentation, again this solution is dissolved in the suitable water solution system, can solve all problems of explaining in the background of invention fully, except methanol, we not yet find can instead of ethanol solvent, but methanol is toxic, can not be as the solvent of injection.

Described buffer also is solvent.As: acetic acid-sodium-acetate buffer, citric acid-sodium citrate buffer, lactic acid-sodium lactate buffer, tartaric acid-sodium tartrate buffer, sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution, dipotassium hydrogen phosphate-potassium phosphate buffer, sodium carbonate-sodium bicarbonate buffer liquid.Preferably phosphoric acid disodium hydrogen-phosphate sodium dihydrogen buffer solution.

Described pH adjusting agent is organic acid or mineral acid.PH adjusting agent is selected from mineral acid or the organic acid of variable concentrations, example hydrochloric acid, lactic acid, citric acid, propanoic acid, formic acid, carbonic acid, hydrochloric acid, tartaric acid, Metaphosphoric acid, sulphuric acid, nitric acid, boric acid, butanoic acid, edetic acid, sorbic acid, salicylic acid, lauric acid, benzoic acid.

Described pH adjusting agent is organic base or inorganic base.PH adjusting agent is selected from organic base or the inorganic base of variable concentrations, as: sodium hydroxide, potassium hydroxide, ammonia, ethylenediamine, diethylamine, triethanolamine, triethylamine.

Described pH adjusting agent is saturated salt solution.As: sodium dihydrogen phosphate, potassium dihydrogen phosphate, sodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium carbonate, sodium bicarbonate, sodium acetate, potassium acetate, sodium tartrate, Soluble tartar., sodium citrate, potassium citrate.

The consumption of described solvent is 5%～50% of cumulative volume, preferred 10%.

The consumption of described solvent buffer is 50%～95% of cumulative volume.

The pH value of described solution is 6.5～8.0.

Another object of the present invention provides a kind of preparation method of Romurtide injection, and the method comprises the steps:

(1) preparation of buffer;

(2) dissolving romurtide, with buffer, adjust pH, depyrogenation;

(3) packing.

Advantage of the present invention is: compare with the injection powder pin, injection need not dissolving, has solved packing and cost of transportation; The injection solution that obtains is clear and bright state fully, and the preparation of existing Japan is rear with certain opalescence with special lyase dissolving, so more convenient, the safety of preparation of the present invention use, and has avoided the possibility of secondary pollution, can save time, improve medical personnel's work efficiency.

Below in conjunction with embodiment the present invention is described in further detail, but the present invention is not subjected to the restriction of these embodiment.

The specific embodiment

Embodiment 1: the preparation Romurtide injection

Pharmaceutical formulation:

Romurtide 0.20g

Ethanol 100ml

Sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution adds to 1000ml

Make 1000

(1) preparation of buffer: take by weighing respectively disodium hydrogen phosphate,anhydrous 8.31g, sodium dihydrogen phosphate dihydrate 2.24g adds water to 1000ml, adds 0.2% (g/v) active carbon, stirs, and leaves standstill 15 minutes, filters carbon removal, solution for standby.

(2) romurtide is dissolved in the ethanol in the prescription ratio, then add sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution, heat 45 ℃, stirred 30 minutes, regulate pH value to 6.5～8.0, then use the ultrafilter depyrogenation, filtrate is measured the content of romurtide in the solution with 0.22 μ m filtering with microporous membrane degerming, determines accurate loading amount, packing is sealed after being filled with nitrogen.

Embodiment 2: the preparation Romurtide injection

Pharmaceutical formulation:

Romurtide 0.20g

1,2-PD 50ml

Ethanol 50ml

Sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution adds to 1000ml

Make 1000

(1) preparation of buffer: take by weighing respectively disodium hydrogen phosphate,anhydrous 8.31g, potassium dihydrogen phosphate 1.95g adds water to 1000ml, adds 0.2% (g/v) active carbon, stirs, and leaves standstill 15 minutes, filters carbon removal, solution for standby.

(2) romurtide is dissolved in ethanol and 1 in the prescription ratio, in the mixed solvent of 2-propylene glycol, then add phosphate buffer, heat 45 ℃, stirred 30 minutes, regulate pH value to 6.5～8.0, then use the ultrafilter depyrogenation, filtrate is with 0.22 μ m filtering with microporous membrane degerming, measure the content of romurtide in the solution, determine accurate loading amount, packing is sealed after being filled with nitrogen.

Embodiment 3: the preparation Romurtide injection

Pharmaceutical formulation:

Romurtide 0.20g

Ethanol 200ml

Acetic acid-sodium-acetate buffer adds to 1000ml

Make 1000

(1) preparation of buffer: take by weighing respectively sodium acetate 6.8g, acetic acid is an amount of, adds water to 1000ml, and making its pH is 6.5-8.0, adds 0.2% (g/v) active carbon, stirs, and leaves standstill 15 minutes, filters carbon removal, solution for standby.

(2) romurtide is dissolved in the ethanol in the prescription ratio, then add acetic acid-sodium-acetate buffer, be heated to 45 ℃, stirred 30 minutes, regulate pH value to 6.5～8.0, then use the ultrafilter depyrogenation, filtrate is measured the content of romurtide in the solution with 0.22 μ m filtering with microporous membrane degerming, determines accurate loading amount, packing is sealed after being filled with nitrogen.

Embodiment 4: the preparation Romurtide injection

Pharmaceutical formulation:

Romurtide 0.20g

Macrogol 200 100ml

Ethanol 100ml

Acetic acid-sodium-acetate buffer adds to 1000ml

Make 1000

(1) preparation of buffer: take by weighing respectively sodium acetate 6.8g, acetic acid is an amount of, adds water to 1000ml, and making its pH is 6.5-8.0, adds 0.2% (g/v) active carbon, stirs, and leaves standstill 15 minutes, filters carbon removal, solution for standby.

(2) be dissolved in romurtide in the mixed solvent of ethanol and Macrogol 200 in the prescription ratio, then add acetic acid-sodium-acetate buffer, be heated to 45 ℃, stirred 30 minutes, regulate pH value to 6.5～8.0, then use the ultrafilter depyrogenation of the molecular weight 5000 that dams, filtrate is measured the content of romurtide in the solution with 0.22 μ m filtering with microporous membrane degerming, determines accurate loading amount, packing is sealed after being filled with nitrogen.

Embodiment 5: the Romurtide injection stability test

One, placement condition

Romurtide solution put respectively under 60 ℃, 40 ℃, 30 ℃, 25 ℃, 4500LX illumination condition placed 10 days, in the 5th, 10 day sampling and measuring.

Two, detection method

1. outward appearance: range estimation

2. related substance: HPLC method

Instrument: Japanese Shimadzu high performance liquid chromatograph

Chromatographic column: C ₁₈Post (150mm * 4.6mm, 5 μ m)

Mobile phase: methanol-acetonitrile-0.02mol/L ammonium acetate buffer (transferring pH4.8 with glacial acetic acid)=18: 60: 22

Detect wavelength: 220nm

Algoscopy: get this product sample solution as need testing solution; Precision is measured need testing solution 1ml, is diluted to 100ml with mobile phase, shakes up, in contrast solution.Precision is measured contrast solution 20 μ l injection liquid chromatographies, regulate detection sensitivity, the peak height that makes the main constituent peak is 15% of full scale, measures need testing solution 20 μ l injection liquid chromatographies again, the record chromatogram calculates related substance to 3 times of the main peak retention time.

3. content:

The same related substance of chromatographic condition

Algoscopy: get this product sample solution as need testing solution; In addition to take by weighing the romurtide reference substance an amount of for precision, makes in contrast product solution of every milliliter of solution that contains 200 μ g with mobile phase dissolving and dilution, measures respectively 20 μ l injection liquid chromatographies, and the record chromatogram is by external standard method, with calculated by peak area content.

Three, testing result

Table 1: hot test result

Table 2: exposure experiments to light result

Placed 10 days under 60 ℃ of conditions as a result, sample appearance is without significant change, and related substance obviously increases, and content obviously descends; Placed 10 days under the condition under 40 ℃ of conditions, sample appearance is without significant change, and related substance slightly increases content significant change does not occur; Placed 10 days under 30 ℃ of conditions, sample appearance is without significant change, and related substance slightly increases, and content slightly descends; Placed 10 days under 25 ℃ of conditions, the every detection index of sample has no significant change, and is consistent with 0 day.Illustrate that this product is unstable to high temperature, should seal in the cool preservation, and in preparation process, also should avoid long-time high temperature more than 45 ℃.

Claims (7)

1. Romurtide injection that comprises the romurtide as active component, solvent, pH adjusting agent and the buffer solvent of effective dose is characterized in that:

Described solvent is selected from ethanol, glycerol, 1,2-PD, low molecular poly or their mixture;

Described buffer solvent is selected from acetic acid-sodium-acetate buffer, citric acid-sodium citrate buffer, lactic acid-sodium lactate buffer, tartaric acid-sodium tartrate buffer, sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution, dipotassium hydrogen phosphate-potassium phosphate buffer, sodium carbonate-sodium bicarbonate buffer liquid.

2. Romurtide injection according to claim 1 is characterized in that described solvent is ethanol.

3. Romurtide injection according to claim 1, the consumption that it is characterized in that described solvent is 5%～50% of cumulative volume.

4. Romurtide injection according to claim 3, the consumption that it is characterized in that described solvent is 10%.

5. Romurtide injection according to claim 1 is characterized in that described buffer solvent is sodium hydrogen phosphate-phosphate sodium dihydrogen buffer solution.

6. Romurtide injection according to claim 1, the pH value that it is characterized in that described solution is 6.5～8.0.

7. the preparation method of Romurtide injection according to claim 1, the method comprises the steps:

(1) preparation of buffer;

(2) dissolving romurtide, with buffer, adjust pH, depyrogenation;

(3) packing.