Hemoglobin type oxygen-carrying nano gel and preparation method and application thereof
The technology of hemoglobin and nanogel is applied in the field of hemoglobin oxygen-carrying nanogel and its preparation, which can solve the problems of high price of phospholipids, complicated process, instability, etc., achieve good application prospects and economic benefits, overcome side effects, and The effect of improving safety
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Embodiment 1
[0074] Embodiment one: the preparation of raw material of the present invention
[0075] 1. Synthesis method of mPEG-PEI 1
[0076] Weigh polyethylene glycol monomethyl ether mPEG (Mw=2000-10000) in a three-neck flask, dissolve it in anhydrous toluene, weigh 10 times the molar amount of isophorone diisocyanate (IPDI) and dissolve it in the toluene solution , add a catalytic amount of stannous octoate, mix the above two solutions, and reflux at 80°C for 8 to 10 hours. The obtained product was precipitated with ice petroleum ether (petroleum ether frozen at -20°C for more than 4 hours), repeated several times, and dried in vacuum at 40°C for 48 hours to obtain mPEG-IPDI for use;
[0077] Then accurately weigh 1.1 times the molar amount of branched PEI (polyethyleneimine) (Mw=600-10000) and dissolve it in anhydrous toluene, and magnetically stir to help dissolve;
[0078] Then the MPEG-IPDI obtained in the previous step reaction was dissolved in toluene solution and transferred...
Embodiment 2
[0092] Example 2 Preparation of hemoglobin / haptoglobin nanogel (hemoglobin oxygen-carrying nanogel) of the present invention
[0093] 1. Preparation of copolymer material mPEG-PEI-g-SH
[0094] Polyethylene glycol monomethyl ether (mPEG-COOH): Mn=2000g / mol
[0095] Branched PEI (AlfaAsea): Mn=600g / mol
[0096] Isophorone diisocyanate (IPDI): Mn=222.28g / mol
[0097] Mercapto polyethylene glycol NHS ester (SH-PEG-NHS): Mn=200g / mol
[0098] The mPEG-PEI-g-SH copolymer material of the present invention was prepared using mPEG-COOH with Mn=2000 and PEI with Mn=1200. See Example 1 for specific reaction conditions. The specific steps are: 1. Synthesize mPEG-PEI according to mPEG-PEI synthesis method 1 in Example 1; 2. Synthesize according to mPEG-PEI-g-SH synthesis method 2 in Example 1 to obtain the nanocarrier material mPEG-PEI-g- sh. The NMR image of the product is as image 3 As shown in a, it shows that it has been successfully synthesized. According to elemental analysis...
Embodiment 3
[0115] Example 3 Preparation of hemoglobin / haptoglobin nanogel (hemoglobin oxygen-carrying nanogel) of the present invention
[0116] 1. Preparation of nanogel mPEG-PEI-g-SH
[0117] Monofunctional polyethylene glycol (mPEG-COOH): Mn=5000g / mol
[0118] Branched PEI (AlfaAsea): Mn=1800g / mol
[0119] 2-iminothiolane (Traut's reagent)
[0120] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl):
[0121] N-Hydroxysuccinimide (NHS):
[0122] The mPEG-PEI-G-SH copolymer material of the present invention was prepared using mPEG-COOH with Mn=5000 and PEI with Mn=1800. See Example 1 for specific reaction conditions. The specific steps are: 1. Synthesize mPEG-PEI according to the mPEG-PEI synthesis method 2 in Example 1; 2. Synthesize according to the mPEG-PEI-g-SH synthesis method 1 in Example 1 to obtain the nanocarrier material mPEG-PEI-g- sh. The NMR image of the product is as image 3 As shown in b, it shows that it has been successfully synthesized. Acco...
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