COMPLEX CONTAINING UNDENATURED TYPE II COLLAGEN AND CHITOSAN OLIGOSACCHARIDE, AND PREPARATION METHOD THEREFOR

Abstract

A complex containing undenatured type II collagen and chitosan oligosaccharide, and a preparation method therefor. The preparation method comprises the following steps: (1) activation: mixing a certain amount of powder or particulate matter containing undenatured type II collagen with an activating agent and storing the mixture at 0° C. to 80° C. for 1 min to 300 min; (2) sieving: sieving the mixture obtained in step (1) to remove unreacted activating agent; (3) coupling: adding chitosan oligosaccharide to the mixture obtained in step (2) in which the unreacted activating agent is removed, and dynamically mixing them at 0° C. to 80° C. for 1 min to 300 min; and (4) drying: drying the mixture obtained in step (3) and controlling the water content to be 8% or less, the resulting product being a complex containing undenatured type II collagen and chitosan oligosaccharide. This preparation method is simple to implement, with fewer operation steps and relatively high efficiency.

Claims

1. A method for preparing a complex containing undenatured type II collagen and chitosan oligosaccharide, comprising the following steps: (1) activation: mixing a certain amount of powder or particulate matter containing undenatured type II collagen with an activating agent and storing the mixture at 0° C. to 80° C. for 1 min to 300 min; (2) sieving: sieving the mixture obtained in step (1) to remove unreacted activating agent; (3) coupling: adding chitosan oligosaccharide to the mixture obtained in step (2) in which the unreacted activating agent is removed, and dynamically mixing them at 0° C. to 80° C. for 1 min to 300 min; (4) drying: drying the mixture obtained in step (3) and controlling the water content to be 8% or less, the resulting product being a complex containing undenatured type II collagen and chitosan oligosaccharide.

2. The preparation method according to claim 1, wherein the activating agent added in the step (1) is flavone, anthocyanidin, polyphenols or plant extracts containing the above substances.

3. The preparation method according to claim 2, wherein the mass ratio of the activating agent to the undenatured type II collagen is 100:1 to 1:10000.

4. The preparation method according to claim 1, wherein the chitosan oligosaccharide added in the step (3) has a degree of polymerization of 2 to 10.

5. The preparation method according to claim 1, wherein the chitosan oligosaccharide added in the step (3) is a mixture of chitosan oligosaccharides with varying degrees of polymerization, with an average degree of polymerization ranging from 2 to 10.

6. The preparation method according to claim 1, wherein the mass ratio of the chitosan oligosaccharide added in the step (3) to the undenatured type II collagen is 100:1 to 1:200.

7. The preparation method according to claim 1, wherein, in the step (2), the mixture obtained in the step (1) is sieved using a screen.

8. A complex containing undenatured type II collagen and chitosan oligosaccharide prepared by the preparation method according to claim 1.

9. The complex containing undenatured type II collagen and chitosan oligosaccharide according to claim 8, wherein the undenatured type II collagen and chitosan oligosaccharide in the complex are present in a chemically bonded form.

10. The complex containing undenatured type II collagen and chitosan oligosaccharide according to claim 9, wherein the ratio of the undenatured type II collagen to chitosan oligosaccharide is in a range of 1:200 to 100:1.

Description

DETAILED DESCRIPTION

[0022] The present application will be described in detail below by specific embodiments. It should be noted that the embodiments are only examples of the complex containing undenatured type II collagen and chitosan oligosaccharide and the preparation method therefor in the present application, which cannot represent the entirety of the patent of this invention and should not be construed as the limitation to the scope of protection for this invention. Some adjustments or improvements can also be made under the premise of not departing from the essential concept of this invention, all of which fall within the scope of protection for this invention.

[0023] According to general concept of the present application, provided is a method for preparing a complex containing undenatured type II collagen and chitosan oligosaccharide, which includes the following steps: [0024] (1) activation: mixing (e.g., evenly) a certain amount of powder or particulate matter containing undenatured type II collagen (e.g., powder or particulate matter obtained from pretreatment of animal cartilages) with an activating agent and storing the mixture at 0° C. to 80° C. (e.g., 35° C. to 40° C., or 50° C. to 55° C.) for 1 min to 300 min (e.g., 90 min or 200 min); [0025] (2) sieving: sieving the mixture obtained in step (1) to remove unreacted activating agent; [0026] (3) coupling: adding chitosan oligosaccharide to the mixture obtained in step (2) in which the unreacted activating agent is removed, and dynamically mixing them at 0° C. to 80° C. (e.g., 35° C. to 40° C., or 50° C. to 55° C.) for 1 min to 300 min (e.g., 90 min or 120 min); [0027] (4) drying: drying the mixture obtained in step (3) and controlling the water content to be 8% or less (e.g., 7% or less, or 6% or less), the resulting product being a complex containing undenatured type II collagen and chitosan oligosaccharide.

[0028] In the preparation method according to embodiments of the present application, chitosan oligosaccharide is immobilized on the surface of undenatured type II collagen by using an activating agent, making the undenatured type II collagen less susceptible to degradation by proteases in the human body after oral administration, thereby enhancing the targeted release ratio of the undenatured type II collagen in the intestines. Additionally, the addition of the activating agent increases the density of positive charges on the surface of the undenatured type II collagen, so the inclusion of undenatured type II collagen in the complex facilitates the phagocytosis of collagen by the cells on the surface of the intestines, thereby improving the phagocytosis rate of the undenatured type II collagen by intestinal cells. The complex obtained by the preparation method of the present application enhances the therapeutic effect of undenatured type II collagen.

[0029] In the embodiments, the activating agent added in the step (1) is flavone, anthocyanidin, polyphenols or plant extracts containing the above substances.

[0030] In the embodiments, the mass ratio of the activating agent to the undenatured type II collagen powder or particulate matter is 100:1 to 1:10000, e.g., 1:10.

[0031] In the embodiments, the chitosan oligosaccharide added in the step (3) has a degree of polymerization of 2 to 10, e.g., 3 or 4.

[0032] In an example, the chitosan oligosaccharide added in the step (3) is a mixture of chitosan oligosaccharides with varying degrees of polymerization, with an average degree of polymerization ranging from 2 to 10, e.g., 3 or 4.

[0033] In an example, the mass ratio of the chitosan oligosaccharide added in the step (3) to the undenatured type II collagen powder or particulate matter is 100:1 to 1:200, e.g., 2.5:1 or 1:5.

[0034] In the embodiments, in the step (2), the mixture obtained in the step (1) is sieved using a screen. For example, the mixture obtained in the step (1) is sieved using a screen of 200 to 500 meshes.

[0035] According to another concept of the present application, also provided is a complex containing undenatured type II collagen and chitosan oligosaccharide prepared by the preparation method in any one of the above embodiments.

[0036] In the embodiments, the undenatured type II collagen and chitosan oligosaccharide in the complex are present in a chemically bonded form.

[0037] In the embodiments, the ratio of the undenatured type II collagen to chitosan oligosaccharide is in a range of 1:200 to 100:1.

[0038] Only a part of the embodiments of the present application are shown below for exemplary illustration; other feasible embodiments of the present application will be understood by those skilled in the art and will not be repeated here.

Embodiment 1

[0039] Particulate matter containing 20% of undenatured type II collagen obtained from pretreatment of animal cartilage was used as the raw material, of which the water content was about 20%, and the activating agent used was soy isoflavone. [0040] (1) Activation: 200 g of chicken cartilage powder with an average particle size of 300 mesh was mechanically mixed with 4 g of soy isoflavone evenly, and the resulting mixture was stored at 35° C. to 40° C. for 200 min. [0041] (2) Sieving: the mixture obtained in step (1) was sieved using a 500-mesh screen and the retentate was collected. [0042] (3) Coupling: 100 g of chitosan oligosaccharide with an average degree of polymerization of 4 was added into the retentate obtained in step (2) and mixed by shaking at 35° C. to 40° C. for 200 min. [0043] (4) Drying: the mixture obtained in step (3) was dried in vacuum at low temperature until its water content reached 7% or less.

[0044] The mixture contained a complex of undenatured type II collagen and chitosan oligosaccharide, in which the chitosan oligosaccharide was covalently coupled to the surface of the undenatured type II collagen. The test results showed that the ratio of undenatured type II collagen to chitosan oligosaccharide in the complex was 22.3:1.

Embodiment 2

[0045] Lyophilized powder of undenatured type II collagen extracted from chicken cartilage was used as the raw material, of which the water content was about 10%, and the activating agent used was blueberry extract containing 20% of dihydromyricetin, of which the water content was about 6%. [0046] (1) Activation: 100 g of lyophilized powder of undenatured type II collagen was mechanically mixed with 10 g of blueberry extract evenly, and the resulting mixture was stored at 50° C. to 55° C. for 90 min. [0047] (2) Sieving: the mixture obtained in step (1) was sieved using a 200-mesh screen and the lyophilized powder of undenatured type II collagen was collected. [0048] (3) Coupling: 20 g of chitosan oligosaccharide with an average degree of polymerization of 3 was added into the lyophilized powder obtained in step (2) and mixed by shaking at 50° C. to 55° C. for 120 min. [0049] (4) Drying: the mixture obtained in step (3) was dried in vacuum at low temperature until its water content reached 6% or less.

[0050] The mixture contained a complex of undenatured type II collagen and chitosan oligosaccharide. The test results showed that the ratio of undenatured type II collagen to chitosan oligosaccharide in the complex was 20.6:1.