JPS587606B2 - Method for producing stable neocarzinostatin powder - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing stable neocarzinostatin powder.

More specifically, the present invention provides a method for drying a neocartenostatin solution, particularly a neocartenostatin-containing solution purified from a neocartenostatin fermentation liquid, by making carbohydrates present, thereby reducing the drying time and subsequent powder state. The present invention relates to a method for producing stable neocarzinostatin powder, which is characterized by stably maintaining neocarzinostatin activity during storage.

Neocarzinostatin is a drug derived from Streptomyces carzi
Molecular weight: 10 isolated as an anticancer active substance against Ehrlichi's ascites cancer from the culture p solution of Ehrlitsu ascites
, 700 acidic proteins, is an anticancer antibiotic that also has an inhibitory effect on Durham-positive bacteria [Journal of Antibiotics]
iotics)tScience(Science)t17
8t875-876 (1972)).

However, because this substance is highly temperature sensitive, it is difficult to maintain a constant quality for a long period of time, and for practical purposes, it is necessary to prevent inactivation during storage.

Therefore, the present inventors investigated Streptomyces carzinostaticus (Streptomyces carzino).
ammonium sulfate salting out, DEA
E-cellulose chromatography, Sephadex G
-50 After adding various compounds to the neocarzinostatin solution purified by chromatography and freeze-drying,
We searched for a stabilizer that would enable long-term storage of neocarzinostatin lyophilized powder by storing it under heating and measuring its residual activity.

As a result, they found that carbohydrates have a stabilizing effect.

As the carbohydrate, monosaccharides or derivatives thereof, disaccharides, oligosaccharides, or polysaccharides are used.

Monosaccharides include glucose, galactose, mannose, and fructose; monosaccharide derivatives include sorbitol and mannitol; disaccharides include maltose, lactose, sucrose, melibiose, and cellobiose; oligosaccharides include raffinose; and polysaccharides include found that dextrin and dextran were effective.

Furthermore, it has been found that among these, the stabilizing effect of disaccharides is particularly remarkable.

Table 1 shows typical thermal stabilizing effects of maltose and sucrose.

Here, the activity of neocarzinostatin was measured by a microbial assay using a cylinder agar plate method using Sarcina lutea as a test bacterium.

One or more of these carbohydrates are used as additives.

2 ml of an aqueous solution containing 2 m9 of neocarzinostatin was placed in a 10 m7 vial, 50 μm or 100 μm of maltose or sucrose was added to dissolve the solution, lyophilized, and stored at 50° C. for 7 days.

The residual activity rate was determined by setting the neocarzinostatin activity before lyophilization as 100%.

The effective amount of carbohydrates added to the neocarzinostatin solution is equal to or more than the amount of neocarzinostatin in the solution, but it is appropriate to add 10 to 100 times the amount.

The pH of the solution is suitably 4 to 9, and the temperature during drying is suitably 50°C or less.

Example Neocarzinostatin 2rnI purified from the culture medium of Streptomyces carcinostaticus. Two 10 ml vials containing an aqueous solution containing 2-
Two 10 ml vials containing 2 ml of dissolved maltose-added neocarzinostatin solution were prepared and freeze-dried at the same time.

One bottle each of unsweetened and maltose-added neocarzinostatin powder was stored at -20°C, and the remaining one bottle was stored at 50°C for 7 days.

The neocarcinostatin activity of these four samples was measured by the cylinder agar plate method using Sartina lutea as the test bacterium, and the residual activity of each sample was determined with the activity before freeze-drying as 100%.

The results are shown in Table 2.

Claims (1)

[Claims] 1. When drying the neocarzinostatin solution, at least one type of carbohydrate is added to the neocarzinostatin solution,
A method for producing a stable neocarzinostatin powder characterized by drying. 2. The manufacturing method according to claim 1, wherein the carbohydrate is a monosaccharide or a derivative thereof, a disaccharide, an oligosaccharide, or a polysaccharide. 3. The production method according to claim 2, wherein the monosaccharide or its derivative is glucose, galactose, mannose, fructose, sorbitol or mannitol. 4 Disaccharides are maltose, lactose, sucrose,
The manufacturing method according to claim 2, wherein the melibiose or cellobiose is melibiose or cellobiose. 5. The manufacturing method according to claim 2, wherein the oligosaccharide is raffinose. 6. The manufacturing method according to claim 2, wherein the polysaccharide is dextrin or dextran. 7. The production method according to claim 1, wherein the amount of carbohydrate added is equal to or greater than the amount of neocarzinostatin in the solution. 8. Claim 1, wherein the amount of carbohydrate added is 10 to 100 times the amount of neocarzinostatin in the solution.
Manufacturing method described in section. 9. The manufacturing method according to claim 1, wherein the pH of the neocarzinostatin solution is maintained at 4 to 6. 10 Reduce the temperature of the neocarzinostatin solution during drying to 50
The manufacturing method according to claim 1, wherein the temperature is maintained below ℃. 11. The manufacturing method according to claim 1, wherein the drying is freeze drying.