Thymosin

Thymosin. A family of heat-stable, polypeptide hormones secreted by the thymus gland. Their biological activities include lymphocytopoiesis, restoration of immunological competence and enhancement of expression of T-cell characteristics and function. They have therapeutic potential in patients having primary or secondary immunodeficiency diseases, cancer or diseases related to aging.

Method 1. Extract and remove protein from pig thymus. Crush the pig thymus (fresh or frozen) and extract twice. Add 2 times the amount of acidic physiological saline each time, adjust the pH to about 3, extract for 4 hours below 10°C, and filter with gauze. Combine the two filtrates, adjust the pH to about 7, keep the water bath at 80°C for 10 minutes, and cool to below 10°C to filter or centrifuge to obtain the extract. Add about 5 times the amount of cold acetone, let it stand for 4 hours, collect the precipitate, and dehydrate it with acetone and ether to obtain light yellow acetone powder. Porcine thymus [acidic saline]→[pH3, 4h×2, 10℃] filtrate→[pH7; 80℃] extract [cold acetone]→[4h] acetone powder acetone precipitation, protein removal, salting out and acetone powder Add pH7, 0.01mol/L CPB to remove protein. Add 6 times the volume of CPB for the first time, stir for 0.5h, centrifuge, and add 4 times the volume of CPB to the residue once more. Combine the two clear liquids, add ammonium sulfate to 25% saturation, adjust the pH to 7, let stand for several hours, and centrifuge to remove precipitation. Adjust the pH of the supernatant to 4.6, add ammonium sulfate to 50% saturation, stir for 0.5h, stand in the refrigerator for several hours, and collect the precipitate by centrifugation. Acetone powder [CPB]→[pH7] clear solution[(NH4)2SO425% saturation]→[pH7]supernatant[(NH4)SO4 50% saturation]→[pH4.6] salting out product dialysis desalination, Chromatographic separation adds 5 times the amount of distilled water to the salted-out product to adjust the pH to 7. First, the cold distilled water was dialyzed to no sulfate ions, and then dialyzed against pH 7, 0.01 mol/L CPB for 10 hours, and the dialysate was centrifuged to obtain the supernatant. Exchange with EDTA-ion exchange cellulose column at pH 6-7, elute with pH 4, 0.02mol/L CPB, add cold acetone for precipitation, filter with No. 3 vertical melting funnel to obtain acetone powder. Depyrogenation, aseptic potting, dissolve acetone powder in pyrogen-free distilled water, adjust pH 6 with 1mol/L hydrochloric acid, add the same amount of ether and stir vigorously for 1h, centrifuge at 1500r/min for 15min, aspirate the middle clear liquid, and make the same amount of clear liquid The ether was washed twice in the same way, and 5 times the amount of acetone was added to the combined clear liquid to precipitate a precipitate to obtain a dry powder of acetone. Determination of pyrogen, preparation containing thymosin 2.5 mg/ml, pH6-7.5, aseptically filled to obtain a light yellow thymosin injection. Acetone powder [distilled water, ether]→[pH6] pyrogen-free powder [sterile filling]→[pH6-7.5] thymosin injection Method two, extract from calf thymus, filter and mince calf thymus, Add 0.5% n-octanol-0.9% sodium chloride solution about 3 times the volume (V/V) for homogenization, then centrifuge at 1500r/min for 30min, and filter the supernatant with gauze to obtain the filtrate. Calf thymus [0.5% n-octanol-0.9% NaCl] → [centrifugal] supernatant [gauze filtration] → filtrate to remove protein. Heat the filtrate to 80℃ for 15min, and centrifuge at 1500r/min for 30min to remove protein. Get the supernatant. Then add (NH4)2SO4 to 25% saturation, the pulp is suction filtered to obtain the filtrate. Supernatant [80℃, 15min]→[Centrifugation]Supernatant [(NH4)2SO4]→[Pulp Suction Filtration] Filtrate salting out, chromatographic separation The filtrate is adjusted to pH 4.0 with 20% acetic acid, and (NH4) is added 2SO4 reached 50% saturation, centrifuged at 1500×g for 30 min, and precipitated. Wash with -15°C acetone until white powder, and vacuum dry the acetone. The powder was dissolved in a 0.1mol/L, pH8.4 Tris-hydrochloric acid solution, centrifuged at 4000×g for 30 minutes, and the supernatant was obtained. The filtrate [acetic acid, (NH4)2SO4]→[pH4.0 centrifugal] precipitation [0.1 mol/L Tris-hydrochloric acid]→[pH8.4] supernatant desalting. The supernatant was desalted by Sephadex G-150 column to pH 8. 4 Phosphate buffer (0.02 mol/L) is eluted, and then desalted by Sephadex G-25 column, eluted with distilled water, and the active peak is collected and freeze-dried to obtain thymosin. Supernatant [Sephadex G-150 column]→Eluent [Sephadex G-25 column]→Thymosin.

Immunity enhancer. Treatment of immunodeficiency diseases such as thymic hypoplasia syndrome, ataxia telangiectasia. It is used domestically to treat patients with recurrent aphthous ulcers, severe leprosy infections, and other immunocompromised patients. Adverse reactions: A few people have local allergic reactions such as urticaria and rash after taking it, and occasionally systemic allergic reactions such as dizziness and fever. Skin test should be done before injection.

Molecular Weight:3051.3
XLogP3:-22
Hydrogen Bond Donor Count:47
Hydrogen Bond Acceptor Count:57
Rotatable Bond Count:109
Exact Mass:3050.5046466
Monoisotopic Mass:3049.5012918
Topological Polar Surface Area:1370
Heavy Atom Count:212
Complexity:6930
Defined Atom Stereocenter Count:30
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes