Method for preparing charge modified lactoferrin

Abstract

The invention discloses a method for preparing charge modified lactoferrin. The method comprises the following operation steps: dissolving 3-hydroxy-phtalic anhydride in dimethyl sulfoxide to prepare a saturated HP solution; dissolving lactoferrin in a 0.1M sodium phosphate solution to prepare a protein solution of 20mg / mL; uniformly mixing the saturated HP solution and the protein solution of 20mg / mL, regulating the pH to 10, standing at 25 DEG C for 1 hour, dialyzing by adopting PBS which has the pH of 7.4, and filtering and sterilizing with a 0.45mu m membrane to obtain the charge modified lactoferrin. The charge modified lactoferrin can be combined with carrageenan to prepare a composition medicine for preventing and treating infection of human papilloma virus of females and preventing and treating gynecological gram-negative bacterial infection of females.

Description

technical field

[0001] The invention relates to the field of preparation of anticancer drugs, in particular to a preparation method of charge-modified lactoferrin. Background technique

[0002] Cervical cancer ranks fourth in the incidence of total cancer in my country, and ranks second in the incidence and mortality of female malignant tumors. Studies have found that HPV can be detected in about 99.7% of cervical cancer patients, among which high-risk HPV is closely related to cervical cancer . Every year, there are more than 500,000 new cases of cervical cancer worldwide, and more than 200,000 women die of cervical cancer. There are nearly 100,000 new cases of cervical cancer in China every year, accounting for nearly 20% of the total number of new cases in the world. Cervical cancer is the first cancer completely caused by HPV infection confirmed by the World Health Organization (WHO). HPV belongs to a group of DNA viruses in subgroup A of Papovaviridae. It is an icosah...

Examples

Embodiment 1

[0032] Example 1: Preparation of charge-modified lactoferrin

[0033] Put 1.65 (0.8-16.5) parts by weight of 3-hydroxy-phthalic anhydride (HP) in a 10mL small beaker, add an appropriate amount of dimethyl sulfoxide (DMSO), and stir at 200-500rpm in a magnetic stirrer, At the same time, DMSO was added dropwise to about 10 (5-100) parts by weight to prepare a saturated HP solution; under stirring at 200-500 rpm, 2 g of lactoferrin was dissolved in 100 mL of 0.1 M sodium phosphate buffer solution to prepare a 20 mg / mL solution. Protein solution; then take saturated HP solution and 20mg / mL protein solution and mix at a ratio of 1:100, stir for 15-30min to mix evenly, and adjust the pH to 10 with phosphoric acid or sodium phosphate, let it stand at 25°C for 1h, and use pH7.4 dialyzed with PBS, and sterilized by 0.45 μm membrane filtration to obtain charge-modified lactoferrin, specifically as figure 1 shown.

Embodiment 2

[0034] Embodiment 2: combination medicine

[0035] The combined medicine consists of 0.01 parts by weight of LK protein (charge-modified lactoferrin), 1.5 parts by weight of carrageenan and auxiliary materials, a total of 100 parts by weight. The auxiliary materials include 5 parts by weight of carbomer, 50 parts by weight of glycerin, 0.1 parts by weight of ethylparaben, the combined medicine is a kind of gel, suppository and dressing.

Embodiment 3

[0036] Embodiment 3: the preparation method of combination medicine

[0037] S1: Weigh each raw material according to the proportion, stir 0.5g carbomer in water at 80°C until fully swollen, add 0.15g carrageenan, stir until completely dissolved, and filter and sterilize through a 0.22μm membrane while hot to obtain 1 No. solution;

[0038]S2: Dissolve 0.01g of ethylparaben in water, then add 5g of glycerol, filter and sterilize through a 0.22μm membrane to obtain No. 2 solution;

[0039] S3: Add No. 2 solution to No. 1 solution under agitation and mix evenly, add 0.8 g of triethanolamine, adjust the pH to 5.5, and obtain No. 3 solution;

[0040] S4: Dissolve 0.001g of LK protein in an appropriate amount of water with low-speed stirring at room temperature, and filter and sterilize through a 0.22 μm membrane to obtain solution No. 4;

[0041] S5: under stirring at a certain temperature, add No. 4 solution into No. 3 solution and mix evenly, and adjust the pH to 5.5 with ster...

technical field

[0001] The invention relates to the field of preparation of anticancer drugs, in particular to a preparation method of charge-modified lactoferrin. Background technique

[0002] Cervical cancer ranks fourth in the incidence of total cancer in my country, and ranks second in the incidence and mortality of female malignant tumors. Studies have found that HPV can be detected in about 99.7% of cervical cancer patients, among which high-risk HPV is closely related to cervical cancer . Every year, there are more than 500,000 new cases of cervical cancer worldwide, and more than 200,000 women die of cervical cancer. There are nearly 100,000 new cases of cervical cancer in China every year, accounting for nearly 20% of the total number of new cases in the world. Cervical cancer is the first cancer completely caused by HPV infection confirmed by the World Health Organization (WHO). HPV belongs to a group of DNA viruses in subgroup A of Papovaviridae. It is an icosah...

Examples

Embodiment 1

[0032] Example 1: Preparation of charge-modified lactoferrin

[0033] Put 1.65 (0.8-16.5) parts by weight of 3-hydroxy-phthalic anhydride (HP) in a 10mL small beaker, add an appropriate amount of dimethyl sulfoxide (DMSO), and stir at 200-500rpm in a magnetic stirrer, At the same time, DMSO was added dropwise to about 10 (5-100) parts by weight to prepare a saturated HP solution; under stirring at 200-500 rpm, 2 g of lactoferrin was dissolved in 100 mL of 0.1 M sodium phosphate buffer solution to prepare a 20 mg / mL solution. Protein solution; then take saturated HP solution and 20mg / mL protein solution and mix at a ratio of 1:100, stir for 15-30min to mix evenly, and adjust the pH to 10 with phosphoric acid or sodium phosphate, let it stand at 25°C for 1h, and use pH7.4 dialyzed with PBS, and sterilized by 0.45 μm membrane filtration to obtain charge-modified lactoferrin, specifically as figure 1 shown.

Embodiment 2

[0034] Embodiment 2: combination medicine

[0035] The combined medicine consists of 0.01 parts by weight of LK protein (charge-modified lactoferrin), 1.5 parts by weight of carrageenan and auxiliary materials, a total of 100 parts by weight. The auxiliary materials include 5 parts by weight of carbomer, 50 parts by weight of glycerin, 0.1 parts by weight of ethylparaben, the combined medicine is a kind of gel, suppository and dressing.

Embodiment 3

[0036] Embodiment 3: the preparation method of combination medicine

[0037] S1: Weigh each raw material according to the proportion, stir 0.5g carbomer in water at 80°C until fully swollen, add 0.15g carrageenan, stir until completely dissolved, and filter and sterilize through a 0.22μm membrane while hot to obtain 1 No. solution;

[0038]S2: Dissolve 0.01g of ethylparaben in water, then add 5g of glycerol, filter and sterilize through a 0.22μm membrane to obtain No. 2 solution;

[0039] S3: Add No. 2 solution to No. 1 solution under agitation and mix evenly, add 0.8 g of triethanolamine, adjust the pH to 5.5, and obtain No. 3 solution;

[0040] S4: Dissolve 0.001g of LK protein in an appropriate amount of water with low-speed stirring at room temperature, and filter and sterilize through a 0.22 μm membrane to obtain solution No. 4;

[0041] S5: under stirring at a certain temperature, add No. 4 solution into No. 3 solution and mix evenly, and adjust the pH to 5.5 with ster...

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