Bioactive protein for preventing and treating human papilloma virus infection as well as preparation method and application thereof

A technology of biologically active protein and human papillomavirus, which is applied in the field of medicine, can solve the problems of affecting the effect of anti-HPV virus, poor modification effect of lactoferrin, and large steric hindrance, so as to prevent the expansion of infection, low cost, The effect of small steric hindrance

Active Publication Date: 2021-06-22
江苏正大天创生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The LK protein used in this patent is to modify lactoferrin by 3-hydroxy-phthalic anhydride. Due to the large steric hindrance of 3-hydroxy-phthalic anhydride and low activity, the modification of lactoferrin The effect is not good, affecting the effect of anti-HPV virus

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of acid anhydrided lactoferrin

[0037] In this example, the preparation of acid-anhydrided lactoferrin includes the following steps: at a temperature of 25°C, dissolving 4-hydroxyphthalic anhydride in dimethyl sulfoxide to obtain a saturated 4-hydroxyphthalic anhydride solution; Dissolve ferritin in 0.1M trisodium phosphate solution to obtain a protein solution with a final concentration of 30 mg / mL; add the above-mentioned 4-hydroxyphthalic anhydride solution into the protein solution, stir and mix, and the final concentration of 4-hydroxyphthalic anhydride in the reaction system It was 60mM, and left to react for 2 hours; to obtain acid anhydride lactoferrin; use 0.45uM filter membrane to filter and sterilize, and then use pH 7.2 phosphate buffer for ultrafiltration replacement purification, and the molecular weight cut-off of the ultrafiltration membrane used is 10KD; 0.45uM microporous membrane filter to sterilize, add the protective agent man...

Embodiment 2

[0039] Embodiment 2 prepares acid anhydride lactoferrin

[0040] Dissolve 4-hydroxyphthalic anhydride in dimethyl sulfoxide at a temperature of 35°C to obtain a saturated 4-hydroxyphthalic anhydride solution; dissolve lactoferrin in 0.1M trisodium phosphate solution to obtain a final concentration of 50mg / mL protein solution; add the above-mentioned 4-hydroxyphthalic anhydride solution into the protein solution, stir and mix, the final concentration of 4-hydroxyphthalic anhydride in the reaction system is 80mM, and let it stand for 2 hours to obtain acid anhydrided lactoferrin; Use a 0.45uM filter membrane to sterilize by suction and then use pH 7.2 phosphate buffer for ultrafiltration replacement purification. The molecular weight cut-off of the ultrafiltration membrane used is 10KD; Use a freeze dryer to freeze-dry, collect the freeze-dried powder, and obtain the acid-anhydrided lactoferrin freeze-dried powder.

[0041] The protein freeze-drying step includes the following ...

Embodiment 3

[0042] Example 3 Preparation of acid anhydrided lactoferrin

[0043] At a temperature of 50°C, 4-hydroxyphthalic anhydride was dissolved in dimethyl sulfoxide to obtain a saturated 4-hydroxyphthalic anhydride solution; lactoferrin was dissolved in 0.1M trisodium phosphate solution to obtain a final concentration of 80mg / mL protein solution; add the above-mentioned 4-hydroxyphthalic anhydride solution into the protein solution, stir and mix, the final concentration of 4-hydroxyphthalic anhydride in the reaction system is 80mM, let it stand for 2 hours to react; obtain acid anhydrided lactoferrin; Use a 0.45uM filter membrane to sterilize by suction and then use a pH 7.2 phosphate buffer for ultrafiltration replacement purification. The molecular weight cut-off of the ultrafiltration membrane used is 10KD; Freeze-drying is carried out by a freeze dryer, and the freeze-dried powder is collected to obtain the freeze-dried powder of acid anhydrided lactoferrin.

[0044] The protei...

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Abstract

The invention provides a bioactive protein for preventing and treating human papilloma virus infection as well as a preparation method and application thereof. The bioactive protein is anhydridized lactoferrin, anhydridized whey protein or anhydridized histone, and the anhydridization is obtained by modifying protein by adopting 4-hydroxyphthalic anhydride, a derivative of the 4-hydroxyphthalic anhydride, succinic anhydride or maleic anhydride. The bioactive protein disclosed by the invention has the advantages of preventing HPV virus from invading cells, preventing virus from expanding infection, resisting bacteria and oxidation, being safe, stable, convenient to store and transport, low in cost and the like.

Description

technical field [0001] The invention relates to a biologically active protein for preventing and treating human papillomavirus infection, a preparation method and application thereof, belonging to the technical field of medicine. Background technique [0002] Human papillomavirus (Human papilloma viruses, HPV) is one of the few viruses that are clearly associated with human cancer, and its persistent infection is an important risk factor for cervical cancer. According to statistics, among all cervical cancer patients, high-risk HPV DNA can be detected in more than 99% of the cases. More than 50% of sexually active women have been infected with HPV in their lifetime, and those with persistent high-risk HPV infection have a 100-300-fold increased risk of cervical cancer. Therefore, early intervention and elimination of HPV infection is very important for the prevention of cervical lesions CIN. At present, there are many treatments for HPV infection, among which topical medic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/54A61K38/40A61K38/17A61K47/32A61P31/20A61P35/00
CPCA61K47/54A61K38/40A61K38/1709A61K47/32A61P31/20A61P35/00A61K9/06A61K9/08A61K47/10A61K9/0034A61K9/02A61P31/00A61K38/018A61K38/17C07K14/79
Inventor 王曙蒙常天安陈海鹏曹丹枫魏赵延徐倩
Owner 江苏正大天创生物工程有限公司
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