5α-androstane-3β,5,6β-triol injection and preparation method therefor

Preparation of 200 Ampoules of YC-6 Liquid Injection (Specification 5 ml: 50 mg)

Formula:

YC-6 10 g

2-hydroxypropyl-β-cyclodextrin 200 g

Sodium chloride 1.25 g

Water for injection qs1000

ml

Preparation: 2-hydroxypropyl-β-cyclodextrin was dissolved in 80% of the fresh water for injection and YC-6 was added. The resulting mixture was stirred at room temperature for 10-20 min to make YC-6 completely dissolved. Sodium chloride was added and dissolved by stirring. Water was added to make the volume to 1000 ml. Activated charcoal (0.1%) was added into the above solution, which was stirred for 15 min at 60° C. and then was allowed to cool down to room temperature. A 0.22 μm microporous membrane filter was used to filter the solution. The filtrate was collected and filled to prepare 5 ml injection, which was then subjected to sterilization at 121° C. for 15 min.

Preparation of 200 Ampoules of YC-6 Liquid Injection (Specification 10 ml: 80 mg)

Formula:

YC-6 16 g

3-hydroxypropyl-β-cyclodextrin 400 g

Glucose 13.9 g

Water for injection qs 2000

ml

Preparation: 3-Hydroxypropyl-β-cyclodextrin was dissolved in 80% of the fresh water 15 for injection and YC-6 was added. The resulting mixture was stirred at room temperature for 10-20 min to make YC-6 completely dissolved. Glucose was added and dissolved by stirring. Water was added to make the volume to 2000 ml. Activated charcoal (0.1%) was added into the above solution, which was stirred for 15 min at 60° C. and then allowed to cool naturally to room temperature. A 0.22 μm microporous membrane filter was used to filter the solution. The filtrate was collected and filled to prepare 10 ml injection, which was then subjected to sterilization at 121° C. for 15 min.

Preparation of 200 Ampoules of YC-6 Liquid Injection (Specification 5 ml: 100 mg)

Formula:

YC-6 20 g

2-hydroxypropyl-β-cyclodextrin 400 g

Water for injection qs 1000

ml

Preparation: 2-Hydroxypropyl-f-cyclodextrin was dissolved in 80% of the fresh water for injection and YC-6 was added. The resulting mixture was stirred at room temperature for 10-20 min to make YC-6 completely dissolved. Water was added to make the volume to 1000 ml. Activated charcoal (0.1%) was added into the above solution, which was stirred for 15 min at 60° C. and allowed to cool naturally to room temperature. A 0.22 μm microporous membrane filter was used to filter the solution. The filtrate was collected and filled to prepare 5 ml injection, which was then subjected to sterilization at 115° C. for 30 min.

Preparation of 200 Ampoules of YC-6 Sterile Powder (Specification 80 mg/Bottle)

Formula:

YC-6 16 g

2-hydroxypropyl-β-cyclodextrin 400 g

Sodium chloride 2.5 g

Packaged into 200 ampoules

Preparation: 2-Hydroxypropyl-β-cyclodextrin was dissolved in 80% of the fresh water for injection and YC-6 was added. The resulting mixture was stirred at room temperature for 10-20 min to make YC-6 completely dissolved. Sodium chloride was added and dissolved by stirring. Water was added to make the volume to 2000 ml. Activated charcoal (0.1%) was added into the above solution, which was stirred for 15 min at 60° C. and then allowed to cool naturally to room temperature. A 0.22 μm microporous membrane filter was used to filter. The filtrate was subjected to spray drying and then packaged into 200 ampoules.

Preparation of 200 Ampoules of YC-6 Freeze Drying Powder (Specification 5 ml: 60 mg)

Formula:

YC-6 12 g

3-hydroxypropyl-β-cyclodextrin 200 g

Glucose 7 g

Water for injection qs1000 ml

Preparation: 3-Hydroxypropyl-β-cyclodextrin was dissolved in 80% of the fresh water for injection and YC-6 was added. The resulting mixture was stirred at room temperature for 10-20 min to make YC-6 completely dissolved. Glucose was added and dissolved by stirring. Water was added to make the volume to 1000 ml. Activated charcoal (0.1%) was added into the above solution, which was stirred for 15 min at 60° C. and allowed to cool naturally to room temperature. A 0.22 μm microporous membrane filter was used to filter the solution. The filtrate was packaged into 5 ml ampoules and then was subjected to freeze drying.

Compatible Stability of YC-6 Injection and Conventional Infusions

Two ampoules of YC-6 injection (5 ml×2) of Example 1 were added to conventional infusions to evaluate the compatible stability of YC-6 therein in 8 h. Compatible stability was evaluated in terms of color, clarity, pH and YC-6 content. Results are shown in the following tables.

Preliminary Evaluation on the Safety of YC-6

Kunming mice were caged by weight and randomly divided into 5 groups. Each group had 10 mice, with half males and half females. The YC-6 injection 10 prepared in Example 3 (20 mg/ml) was i.v. administrated via tail vein at different doses. All mice were sacrificed after one-week observation. The toxic reaction and number of death of the animals were recorded every day. LD 50 and 95% confidence were calculated. LD 50 of YC-6 was more than 400±121 mg/kg.

Blood cells were prepared from fresh blood obtained from New Zealand rabbits according to conventional methods. The blood cells were diluted with saline to a 2% suspension. The YC-6 injection prepared in Example 1 was then added to the 2% suspension and incubated at 37° C. for 3 hours. Hemolysis rate was determined by using colorimetric method. Hemolysis rate of YC-6 injection was less than 1%.

Albino guinea pigs were subjected to anaphylactic test according to conventional procedures. No anaphylactic reaction was observed after intravenous administration of the YC-6 injection prepared in Example 1.

New Zealand rabbits were used to perform vascular stimulation test of intravenous administration of the YC-6 injection prepared in Example 1. The results showed that tissue changes of ear edge vein were similar between treatment group and control group. Each rabbit had integral vascular wall of ear edge vein and normal vein structure. No pathological change such as endothelial cells damage or surrounding tissue edema was observed.