| Name | Streptodornase/streptokinase |
| Synonyms | SK-SD Dornokinase Varidase (Lederle) Mixture with streptokinase strptokinase-strrptodornase |
| CAS | 8048-16-6 |
| Physical and Chemical Properties | The double-stranded enzyme system is a mixed enzyme preparation prepared by extracting and separating from the β hemolytic Streptococcus culture medium, and is a white, off-white crystalline or amorphous powder. Soluble in water. However, the aqueous solution is rapidly inactivated at room temperature, 2 ~ 100C can be stable for 7 days, and has the maximum activity in neutral medium. The powder can maintain the activity for 18 months below 40C. Streptokinase dissolves the fibrous portion of thrombus and exudate, and deoxyribonucleic acid hydraulic the viscous nucleoprotein of dead cells. The combination of the two enzymes can promote the removal of blood clots and fibrous or purulent deposits after trauma or inflammation. |
| chemical properties | the mixed enzyme preparation prepared by extracting and separating the double-stranded enzyme system from the β-hemolytic streptococcus culture solution is white, white-like crystalline or amorphous powder. Soluble in water. However, the aqueous solution is rapidly inactivated at room temperature, stable for 7 days at 2 ~ 100C, and has the greatest activity in neutral medium. The powder can maintain activity for 18 months below 40C. Streptokinase can dissolve the fibrous part of thrombus and exudate, and deoxyribonucleic acid can hydrolyse the viscous nuclear protein of dead cells. The combination of the two enzymes can promote the removal of blood clots and fibers or purulent deposits after trauma or inflammation. |
| use | double-stranded enzyme can be used as an auxiliary drug for the treatment of empyema, hemothorax, hematoma, drainage sinus chronic suppuration, osteomyelitis, ulcer and infectious trauma. It is also used for anterior chamber hemorrhage and vitreous hemorrhage. However, intracavitary injection can cause fever and bleeding. Fever can be treated with glucocorticoid stimulation at least. Hemorrhage is effective with 6-amino hexanoic acid. This product is an adjuvant therapeutic drug. It can only be applied on the basis of antibacterial treatment, surgical debridement and drainage. This product is contraindicated in patients with acute suppurative peak fossa tissue inflammation, active tuberculosis and bronchopleural fistula to avoid the spread of the lesion. Low fibrinogen, low fibrinogen, liver function, and blood abnormalities are also prohibited. This product can be injected into the affected part of the cavity or used locally. Intracavitary injection, for hemothorax and empyema, can be 100,000 unit SK and 50,000 unit SD dissolved in more than 10ml of normal saline injection. For maxillary sinus empyema, 10,000~15,000 units of SK and 5000~7500 units of SD (dissolved in 2~3ml of normal saline) can be injected. Mouth, 4 times a day, 1 tablet each time, for 4-6 days, dripping, local dripping with 1000 units/ml solution, '1-2h1 times, local injection, intraocular posterior or subconjunctival injection, injection around the lesion, the concentration is 1000-2000 units/ml. |
| production method | strain culture β hemolytic streptococcus was inoculated into 5ml of beef soup strain medium containing 2% sheep serum, and the pH was adjusted to 7.1~7.2, and 370C was cultured for about 8 hours. When the culture medium is turbid (otherwise the culture time should be prolonged), the strain is transplanted on an agar plate and cultured for 16~18h, which is the production strain. It can be stored in a 40C refrigerator and can be used for 1 month. Strain beef soup medium; 370;17h → strain culture plate colony 1 to 3 seed culture plate colony 5ml seed medium, under 370C, culture for 5~8h. When receiving tertiary seeds, the seed quantity is 2% ~ 3% in a 10L seed tank, and the tertiary seed culture solution can be obtained after 10~13h of 370C culture. Plate colony medium; 370;6h → {primary seed culture solution primary seed culture solution medium; 370;6h → secondary seed culture secondary seed culture solution medium; 370;11h → tertiary seed culture tertiary seed culture fermentation culture 150L of fermentation medium, inoculation amount of 5% ~ 10%, under tank pressure of 19.62kPa (0.2kg), the culture temperature drops to about 330, sodium carbonate (3mol/L) was used to adjust the pH every half an hour to maintain the acidity between pH7.3 and 7.4. The fermentation culture was carried out for about 12 hours. Sampling was carried out to measure the activity unit of chain kinase until the activity of chain kinase reached its peak. The fermentation was stopped for about 14-16 hours. The yield was 519U/ml of chain kinase and 1677U/ml of deoxyribonuclease. Three-stage seed culture medium → fermentation broth adsorption and elution fermentation broth adjust pH7.5 ~ 8 with sodium hydroxide solution (100g/L or 10%), then slowly add 1ml of cholesterol ethanol solution (5%) per 100ml of fermentation broth, and fully stir for 5min. Then add 724 resin that has passed more than 80 mesh into every 100ml of fermentation broth. After the foam disappears, adjust pH4.8 ~ 5 with acetic acid solution (10%). Stir continuously to make it adsorb for 30min, let stand for 15min to discard the supernatant. The resin is cleaned twice with tap water with pH4.8 ~ 5, washed once with distilled water with pH4.8 ~ 5, filtered and drained, transferred into the container, and added a small amount of ice distilled water to be able to stir. Below 15 0C, sodium hydroxide (100g/L or 10%) is added dropwise while stirring until the PH reaches 7.2. The Brinell funnel is filtered, and the resin is rinsed once with a small amount of distilled water, and the filtrate and lotion are combined to obtain the eluent. Cholesterol ethanol liquid from fermentation broth; 724 resin → adsorption of adsorbate H2OnaOH;pH7 → salting out of elution eluent Add 243g of ammonium sulfate per kilogram of eluent to make the saturation of ammonium sulfate reach 40%, adjust ph7 ~ 7.2 with sodium hydroxide (100g/L) at any time, dissolve all ammonium sulfate at about 100C, centrifuge to obtain [I] precipitate for preparing streptokinase; [II] Mother liquor for preparing deoxyribonucleoenzyme, then process separately. eluent (NH4)2SO4;100C → salting out → [I] precipitation (for streptokinase) → mother liquor [II] (for deoxyribonuclease) precipitation-preparation of streptokinase calcium phosphate treatment, precipitation Mash the precipitate, add a certain amount of distilled water and adjust pH7.2 to dissolve it. After dissolution, adjust pH8 ~ 9 with sodium hydroxide (100g/L), under full stirring, add 10~17ml of sodium phosphate (152g/L, I .e. 15.2%) per 100ml of solution (volume is 1/10~1/6 of the solution), add 5~8ml of calcium acetate (226g/L, I .e. 22.6%), centrifuge, collect clarified liquid, and adjust pH7.2 with hydrochloric acid (10%) to obtain enzyme solution. Place the enzyme solution in a salt ice bath on the right and left of 00C, slowly add 30ml of ice methanol to every 100ml of enzyme solution, at this time the temperature should be lower than 80C, then adjust the pH to 5.2~5.4 with hydrochloric acid (10%), and let stand for 15~30min. Centrifuge, collect the precipitate, and obtain the refined product of the enzyme. [I] Precipitation of H2O;Na3PO4;Ca(Ac)2 → Calcium Phosphate Treatment Enzyme Solution Methanol; pH5.3 → Precipitate crude product precipitation isoelectric point treatment, precipitate crude product add appropriate amount of distillate and adjust pH to 7.2., Make it completely dissolved. In ice solution, hydrochloric acid (1mol/L) is used to adjust the ph to 5, precipitation is carried out, precipitation is collected, distilled water with the same volume as the stock solution and pH 7.2 is added, and the temperature is cooled to 00C in an ice bath. Under the condition of less than 50C, 30ml of ice methanol is gradually added to every 100ml of ice solution, the ph is adjusted to 5.4~5.5 with hydrochloric acid (1mol/L), and the temperature is set to stand for 15~30min, centrifuge, get the precipitate. Crude product precipitation distilled water; pH7.2 → enzyme crude solution methanol; pH5.4 → pre-adsorption of precipitation precipitate and isoelectric point treatment dissolve the precipitate in distilled water with pH6.6, give the concentration of enzyme solution 50~1 million units/ml, add the same amount of pH6.6 phosphoric acid buffer (0.2mol/L), and then add 200g of DEAE-cellulose balanced with ph6.6 phosphoric acid flushing solution per 0.3 billion unit/ml of enzyme solution, fully stir right and left for 15min, filter, DEAE-fiber is washed twice with pH 6.6 phosphate buffer (0.1mol/L), filtrate and lotion are combined, pH 5 precipitate is adjusted with hydrochloric acid (1mol/L), streptokinase precipitate is collected by centrifugation, and a distilled water solution with pH 7.2 with enzyme concentration of 50-100U/ml is prepared to obtain refined enzyme solution. DEAE-cellulose column of precipitate; pH6.6 → washing filtrate HCl; Distilled water pH5.5 → 7.2 → enzyme solution adsorption and elution are calculated and fed with 800g of right-left DEAE-cellulose (for refining) per 100 million units, an appropriate amount of pH6.6 phosphoric acid buffer (0.01mol/L) is added, stirred evenly, the enzyme liquid is adsorbed through a chromatographic column, and after completion, a small amount of ph6.6 phosphoric acid buffer (0.01mol/L) is used to wash, elution was carried out with ph6.6 phosphate buffer (0.08mol/L), and streptokinase eluate above 20000U/ml was collected. Enzyme solution DEAE- |