HSCGF liposome as well as preparation and application thereof

Abstract

The invention discloses an HSCGF (Human Stem Cell Growth Factor) liposome as well as the preparation and the application thereof. The HSCGF liposome comprises 2.0 to 4.0 percent of HSCGF, 8.0 to 10.0 percent of phosphatide, 2.5 to 3.5 percent of cholesterol and 10 to 25 percent of protecting agents. The HSCGF is prepared into liposome, the activity of the HSCGF is kept effectively, the stability of SCGF (Stem Cell Growth Factor) can be improved obviously, skin can be promoted to absorb and utilize the SCGF, and the HSCGF liposome has the advantages of good affinity, no toxicity or side effects, and wide application range. The anti-wrinkle performance of cosmetics containing the HSCGF coated with the liposome can still be kept well after the stability is examined for three months. The HSCGF liposome is used as raw material and other active materials and auxiliary materials are used as supplements so as to prepare frequently-used medicine forms to illustrate the drug effect of the SCGF, so that the bioavailability can be improved.

Description

technical field

[0001] The invention belongs to the field of pharmaceutical preparations and cosmetics, and particularly relates to a human stem cell growth factor liposome and a preparation method and application thereof. Background technique

[0002] Human stem cell growth factor (hSCGF, also known as LSLCL) is an early hematopoietic stimulating factor that acts in the bone marrow hematopoietic microenvironment. It has a variety of biological activities and is mainly distributed in the brain, liver, bone marrow, Placenta, testis, kidney, etc., are produced from stromal cells, fibroblasts, endothelial cells, egg cells, podocytes, hepatocytes in these tissues (especially bone marrow). Its gene is located on human chromosome 19q13.3 and is assembled by 4 exons and 3 introns. It is known that human stem cell growth factor has two forms, the full-length molecule hSCGF-α (mature mRNA complete open reading frame 972bp, encoding 323 amino acids, the expressed protein size is abou...

Examples

Embodiment 1

[0036] Weigh 1 g of dry soybean lecithin, 0.1 g of vitamin E and 0.35 g of cholesterol into an eggplant-shaped flask, dissolve with 5-20 ml of absolute ethanol, place the eggplant-shaped flask in a 50°C constant temperature water bath for decompression and rotary evaporation, and pump to absolute ethanol It was evaporated to form a uniform film, and 10 ml of phosphate buffer solution (pH 6.8, 10 mmol / L) containing 400 mg SCGF (hSCGF-α, purchased from Peprotech) and 24% sucrose in mass volume ratio (pH 6.8, 10 mmol / L) was added to rotate and wash the membrane, and finally shake The solution is hydrated for 30 to 60 minutes to obtain SCGF phospholipid suspension (for human stem cell growth factor liposome).

Embodiment 2

[0038] Weigh 0.9g of dry soybean lecithin, 0.1g of vitamin E and 0.3g of cholesterol into an eggplant-shaped flask, dissolve with 5-20ml of absolute ethanol, place the eggplant-shaped flask in a 50°C constant temperature water bath, reduce pressure and rotate to evaporate, and pump to anhydrous The ethanol was evaporated to form a uniform film, and 10ml of phosphate buffer solution (pH 6.8, 10mmol / L) containing 300mg SCGF (hSCGF-α, purchased from Peprotech) and 10% sucrose in mass volume ratio (pH 6.8, 10mmol / L) was added to spin the membrane, and the membrane was hydrated. The solution was passed through a high pressure homogenizer (100 MPa, 5 times) to obtain a translucent liposome colloid solution (for human stem cell growth factor liposomes) with a particle size ranging from 100 to 200.

Embodiment 3

[0040]Accurately weigh 800 mg of phospholipids and 250 mg of cholesterol, put them in a 25 ml eggplant-shaped bottle, add chloroform to dissolve and dilute to 10 ml, inject nitrogen, and sonicate at 100 W on an ice bath for 10 min, every 30 s, for a total of 10 ultrasonic waves. Sonicated liposomes were prepared. Then add chloroform equivalent to 2 to 6 times the volume of ultrasonic liposomes to dissolve it, rotate in a water bath at 35 to 45 °C, remove all organic solvents, and then add 200 mg of SCGF (hSCGF-α, purchased from Peprotech) , 2.0g dextran and 1.5g sucrose in phosphate buffered saline (pH 6.8, 10mmol / L) appropriate amount, fill with nitrogen, hydrate for 12h in an ice bath, and intermittently sonicate for 10min (ultrasonic power 100W, sonicated once every 30s) . After ultrafiltration with a 0.22 μm microporous membrane, 10% mannitol was added in a mass-to-volume ratio, vacuum freeze-dried, and the lipid was rehydrated with an appropriate amount of phosphate buff...

technical field

[0001] The invention belongs to the field of pharmaceutical preparations and cosmetics, and particularly relates to a human stem cell growth factor liposome and a preparation method and application thereof. Background technique

[0002] Human stem cell growth factor (hSCGF, also known as LSLCL) is an early hematopoietic stimulating factor that acts in the bone marrow hematopoietic microenvironment. It has a variety of biological activities and is mainly distributed in the brain, liver, bone marrow, Placenta, testis, kidney, etc., are produced from stromal cells, fibroblasts, endothelial cells, egg cells, podocytes, hepatocytes in these tissues (especially bone marrow). Its gene is located on human chromosome 19q13.3 and is assembled by 4 exons and 3 introns. It is known that human stem cell growth factor has two forms, the full-length molecule hSCGF-α (mature mRNA complete open reading frame 972bp, encoding 323 amino acids, the expressed protein size is abou...

Examples

Embodiment 1

[0036] Weigh 1 g of dry soybean lecithin, 0.1 g of vitamin E and 0.35 g of cholesterol into an eggplant-shaped flask, dissolve with 5-20 ml of absolute ethanol, place the eggplant-shaped flask in a 50°C constant temperature water bath for decompression and rotary evaporation, and pump to absolute ethanol It was evaporated to form a uniform film, and 10 ml of phosphate buffer solution (pH 6.8, 10 mmol / L) containing 400 mg SCGF (hSCGF-α, purchased from Peprotech) and 24% sucrose in mass volume ratio (pH 6.8, 10 mmol / L) was added to rotate and wash the membrane, and finally shake The solution is hydrated for 30 to 60 minutes to obtain SCGF phospholipid suspension (for human stem cell growth factor liposome).

Embodiment 2

[0038] Weigh 0.9g of dry soybean lecithin, 0.1g of vitamin E and 0.3g of cholesterol into an eggplant-shaped flask, dissolve with 5-20ml of absolute ethanol, place the eggplant-shaped flask in a 50°C constant temperature water bath, reduce pressure and rotate to evaporate, and pump to anhydrous The ethanol was evaporated to form a uniform film, and 10ml of phosphate buffer solution (pH 6.8, 10mmol / L) containing 300mg SCGF (hSCGF-α, purchased from Peprotech) and 10% sucrose in mass volume ratio (pH 6.8, 10mmol / L) was added to spin the membrane, and the membrane was hydrated. The solution was passed through a high pressure homogenizer (100 MPa, 5 times) to obtain a translucent liposome colloid solution (for human stem cell growth factor liposomes) with a particle size ranging from 100 to 200.

Embodiment 3

[0040]Accurately weigh 800 mg of phospholipids and 250 mg of cholesterol, put them in a 25 ml eggplant-shaped bottle, add chloroform to dissolve and dilute to 10 ml, inject nitrogen, and sonicate at 100 W on an ice bath for 10 min, every 30 s, for a total of 10 ultrasonic waves. Sonicated liposomes were prepared. Then add chloroform equivalent to 2 to 6 times the volume of ultrasonic liposomes to dissolve it, rotate in a water bath at 35 to 45 °C, remove all organic solvents, and then add 200 mg of SCGF (hSCGF-α, purchased from Peprotech) , 2.0g dextran and 1.5g sucrose in phosphate buffered saline (pH 6.8, 10mmol / L) appropriate amount, fill with nitrogen, hydrate for 12h in an ice bath, and intermittently sonicate for 10min (ultrasonic power 100W, sonicated once every 30s) . After ultrafiltration with a 0.22 μm microporous membrane, 10% mannitol was added in a mass-to-volume ratio, vacuum freeze-dried, and the lipid was rehydrated with an appropriate amount of phosphate buff...

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