Method of Treatment or Prevention of Hair Loss or for the Enhancement of Hair Growth
a hair loss and hair growth technology, applied in the field of hair loss treatment or prevention or hair growth enhancement, can solve the problems of uncontrollable hair cycle, inability to easily pull out, anchorage of anagen hair, etc., and achieve the effect of enhancing hair growth or initiation of hair growth, and easy application to the skin
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example 1
Method for Producing Recombinant Midkine
[0132]A cDNA fragment comprising human MK open reading frame (nucleotide positions 1-432; SEQ ID NO: 4) was inserted into the yeast expression vector pPIC9 (Invitrogen). This recombinant plasmid was transfected into yeast (Pichia pastoris GS115; Research Corporation Technologies), and the desired clones were selected with histidine and G418.
[0133]Human MK protein secreted by yeast into the culture medium was purified by employing the following column chromatography in the order below:[0134]1. SP Steamlines (Pharmacia; adsorption and wash with 20 mM pH 5.5 acetate buffer, elution with 20 mM pH 3.5 acetate suffer containing NaCl);[0135]2. Sulfated Cellulofine (Seikagaku Kogyo, Japan; adsorption with 10 mM pH 7.2 phosphate buffer, wash with buffer containing 0.7 M NaCl, elution with buffer containing 2.0 M NaCl);[0136]3. Superdex 75 pg (Pharmacia; gel filtration with saline);[0137]4. Poly. Sulfoethyl A (Poly LC Co.; adsorption with 20 mM buffer c...
example 2
[0140]Hair-growing effects of external applications of midkine (MK) and pleiotrophin (PTN). 1 μg / ml of MK or PTN in 40% glycerol / phosphate buffered saline (PBS) (v / v) was prepared. 10 μl of the solution of MK or PTN was directly applied three times per week to alopecic regions of aged Black 6 mice (1.5 years old, MK heterozygous knockout (+ / −)). It was observed for 25 days after starting the application whether each of MK and PTN had a hair-growing effect. FIG. 1 shows the results of the observation. As clearly understood from FIG. 1 and the brief description thereof, application of MK or PTN promoted hair growth on the alopecic regions of the aged mice. In this experiment, the applications of MK and PTN stopped on Day 15, because the hair-growing effect of MK and PTN was already confirmed on Day 15. From Day 15 to Day 25, the amount of hair increased even though the applications of MK and PTN were stopped (FIG. 1). This shows that the hair-growing effects of MK and PTN last for som...
example 3
Hair-Growing Effects of Subcutaneous Injection of MK and PTN
[0141]1 μg / ml of MK or PTN in PBS was prepared. 50 μl of the solution of MK or PTN was injected subcutaneously into aged Black 6 mice (1.5 years old, MK homozygous knockout (− / −)). The subcutaneous injections of MK and PTN showed no hair-growing effect on Day 15 (FIG. 2) under these conditions in this animal model. After changing on Day 15 from the subcutaneous injection to external applications of MK or PTN in accordance with Example 2, the hair-growing effects of MK or PTN were observed on Day 25 (FIG. 2).
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