JOINT CAVITY INJECTION PREPARATION AND USE THEREOF

Abstract

A joint cavity injection preparation is provided. The active ingredient of the joint cavity injection preparation is deacetylated xanthan gum (XG). The deacetylated XG has a molecular weight of 500,000 to 20,000,000. The joint cavity injection preparation is prepared from deacetylated xanthan gum (XG), which has higher biocompatibility and safe wide-dosage range than existing joint cavity injection preparations prepared from XG.

Claims

1. A joint cavity injection preparation, wherein an active ingredient of the joint cavity injection preparation is deacetylated xanthan gum (XG); the deacetylated XG has a molecular weight of 500,000 to 10,000,000; and a percentage of a mass of the deacetylated XG in a volume of the joint cavity injection preparation is 0.5% to 8% (w/v).

2. The joint cavity injection preparation according to claim 1, wherein the percentage of the mass of the deacetylated XG in the volume of the joint cavity injection preparation is 1% to 5% (w/v).

3. The joint cavity injection preparation according to claim 1, wherein the deacetylated XG has a molecular weight of 800,000 to 3,000,000.

4. The joint cavity injection preparation according to claim 1, further comprising one or more selected from the group consisting of sodium hyaluronate (SH), chondroitin sulfate (CS), chitosan, and trehalose.

5. The joint cavity injection preparation according to claim 1, wherein the joint cavity injection preparation has a pH of 5.5 to 9 and an osmotic pressure of 200 mOsmol/L to 400 mOsmol/L.

6. Use of deacetylated XG in a preparation of a drug for treating osteoarthritis (OA), wherein the deacetylated XG has a molecular weight of 500,000 to 10,000,000; the deacetylated XG is used at an amount of 0.5% to 8% (w/v), and the drug has a dosage form of a joint cavity injection preparation.

7. The use of the deacetylated XG in the preparation of the drug for treating the OA according to claim 6, wherein the drug has higher biosafety than a drug for treating the OA prepared from XG.

8. The use of the deacetylated XG in the preparation of the drug for treating the OA according to claim 6, wherein the deacetylated XG is used at an amount of 1% to 5% in the drug.

9. The use of the deacetylated XG in the preparation of the drug for treating the OA according to claim 6, wherein the deacetylated XG has a molecular weight of 800 000 to 3,000,000.

10. (canceled)

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0025] In order to clearly explain the technical features of the solution, the present disclosure will be described in detail below through specific examples.

EXAMPLE 1

[0026] A Preparation Method of Deacetylated XG was Provided:

[0027] 10 g of XG with a molecular weight of 3,000,000 was taken and fully dissolved in 1,000 mL of water, a pH was adjusted to 11, and a resulting solution was stirred at 50° C. for 0.5 h; a pH was adjusted to 2.0, and a resulting mixture reacted at 0.1 MPa for 20 min and then cooled; a pH was adjusted to 7.0, and precipitation was conducted with ethanol; and a precipitate product was dried at 40° C. under reduced pressure for 24 h to obtain the deacetylated XG.

[0028] According to multi-angle laser test, the deacetylated XG had Mw of 1,000,000, and according to hydroxylamine hydrochloride colorimetry, there were no acetyl groups in a structure of the deacetylated XG.

EXAMPLE 2

[0029] A Preparation Method of Deacetylated XG was Provided:

[0030] 20 g of XG with a molecular weight of 3,000,000 was taken and fully dissolved in 1,000 mL of water, a pH was adjusted to 11, and a resulting solution was stirred at 60° C. for 2 h; a pH was adjusted to 3.0, and a resulting mixture reacted at 0.1 MPa for 30 min and then cooled; a pH was adjusted to 7.0, and precipitation was conducted with ethanol; and a precipitate product was dried at 40° C. under reduced pressure for 24 h to obtain the deacetylated XG.

[0031] According to multi-angle laser test, the deacetylated XG had Mw of 2,000,000, and according to hydroxylamine hydrochloride colorimetry, there were no acetyl groups in a structure of the deacetylated XG.

EXAMPLE 3

[0032] A Preparation Method of Deacetylated XG was Provided:

[0033] 30 g of XG with a molecular weight of 5,000,000 was taken and fully dissolved in 1,000 mL of water, a p1-1 was adjusted to 12, and a resulting solution was stirred at 50° C. for 1 h; a pH was adjusted to 5.0, and a resulting mixture reacted at 0.1 MPa for 20 min and then cooled; a pH was adjusted to 7.0, and precipitation was conducted with ethanol; and a precipitate product was dried at 40° C. under reduced pressure for 24 h to obtain the deacetylated XG.

[0034] According to multi-angle laser test, the deacetylated XG had Mw of 4,000,000, and according to hydroxylamine hydrochloride colorimetry, there were no acetyl groups in a structure of the deacetylated XG.

EXAMPLE 4

[0035] A Preparation Method of Deacetylated XG was Provided:

[0036] 50 g of XG with a molecular weight of 5,000,000 was taken and fully dissolved in 1,000 mL of water, a pH was adjusted to 11, and a resulting solution was stirred at 55° C. for 0.5 h; a pH was adjusted to 7.0, and a resulting mixture reacted at 0.1 MPa for 20 min and then cooled; a pH was adjusted to 7.0, and precipitation was conducted with ethanol; and a precipitate product was dried at 40° C. under reduced pressure for 24 h to obtain the deacetylated XG.

[0037] According to multi-angle laser test, the deacetylated XG had Mw of 5,000,000, and according to hydroxylamine hydrochloride colorimetry, there were no acetyl groups in a structure of the deacetylated

EXAMPLE 5

[0038] A Preparation Method of Deacetylated XG was Provided:

[0039] 50 g of XG with a molecular weight of 8,000,000 was taken and fully dissolved in 1,000 mL of water, a pH was adjusted to 11, and a resulting solution was stirred at 55° C. for 0.5 h; a pH was adjusted to 2.0, and a resulting mixture reacted at 0.1 MPa for 30 min and then cooled; a pH was adjusted to 7.0, and precipitation was conducted with ethanol; and a precipitate product was dried at 40° C. under reduced pressure for 24 h to obtain the deacetylated XG.

[0040] According to multi-angle laser test, the deacetylated XG had Mw of 2,600,000, and according to hydroxylamine hydrochloride colorimetry, there were no acetyl groups in a structure of the deacetylated XG.

[0041] The deacetylated XG products prepared by the above methods were used to prepare joint cavity injection preparations:

EXAMPLE 6

[0042] Joint Cavity Injection Preparation with 1% Deacetylated XG

TABLE-US-00001 Deacetylated XG (with an average molecular 5 g weight of 1,000,000) DSP 3.78 g MSP 0.8 g Sodium chloride 2.055 g WFI add to 500 mL pH 7.4 Osmotic pressure 300 mOsmol/L

EXAMPLE 7

[0043] Joint Cavity Injection Preparation with 3% Deacetylated XG

TABLE-US-00002 Deacetylated XG (with an average molecular 15 g weight of 1,000,000) DSP 3.00 g MSP 0.6 g Sodium chloride 1.65 g WFI add to 500 mL pH 7.4 Osmotic pressure 300 mOsmol/L

EXAMPLE 8

[0044] Joint Cavity Injection Preparation with 5% Deacetylated XG

TABLE-US-00003 Deacetylated XG (with an average molecular 25 g weight of 1,000,000) DSP 2.5 g MSP 0.5 g Sodium chloride 1.36 g WFI add to 500 mL pH 7.4 Osmotic pressure 300 mOsmol/L

EXAMPLE 9

[0045] Joint Cavity Injection Preparation with 1% Deacetylated XG

TABLE-US-00004 Deacetylated XG (with an average molecular 5 g weight of 1,000,000) DSP 3.0 g MSP 0.7 g Sodium chloride 1.60 g WFI add to 500 mL pH 7.4 Osmotic pressure 300 mOsmol/L

EXAMPLE 10

[0046] Joint Cavity Injection Preparation with 5% Deacetylated XG

TABLE-US-00005 Deacetylated XG (with an average molecular 25 g weight of 1,000,000) DSP 2.4 g MSP 0.6 g Sodium chloride 1.20 g WFI add to 500 mL pH 7.4 Osmotic pressure 300 mOsmol/L

EXPERIMENTAL STUDY

[0047] 1. Animal Experiments to Study the Treatment Effects of the Above Deacetylated XG Joint Cavity Injection Preparations on OA:

[0048] 64 New Zealand big-eared white rabbits were randomly divided into 8 groups, each with 8 rabbits (half male and half female): normal control group, negative control group (normal saline (NS)), deacetylated XG group 1 (Example 6, 1%, 1,000,000), deacetylated XG group 2 (Example 7, 3%, 1,000,000), deacetylated XG group 3 (Example 8, 5%, 1,000,000), deacetylated XG group 4 (Example 9, 1%, 5,000,000), ordinary XG group (1%, 1,000,000), and SH control group.

[0049] The left knee ligament was cut by anterior cruciate ligament transection (ACLT) to establish knee OA models, and after the models were successfully established, the joint cavities were injected with the corresponding drugs or INS at a dosage of 0.3 mL/joint. The SH control group was administered once a week, with a total of 5 administrations; and the deacetylated XG groups and the ordinary XG group were administered once every 5 weeks, with a total of 2 administrations. Animals in each group were sacrificed 10 weeks after the first administration, the articular cartilage was subjected to pathological observation and scored, and scoring results were shown in Table 1. The results showed that, when administered once every 5 weeks, both deacetylated XG and ordinary XG exhibited significant therapeutic effects on OA, which had no significant difference from the SH group administered once a week. The pathological scores of articular cartilage were shown in Table 1.

TABLE-US-00006 TABLE 1 Pathological scores of articular cartilage Group Pathological score Normal group 0.82 ± 0.05 Negative control group 10.26 ± 0.64  SH group 4.89 ± 0.32 Ordinary XG group 4.91 ± 0.18 (1%, 1,000,000) Deacetviated XG group 1 4.87 ± 0.42 (1%, 1,000,000) Deacetyiated XG group 2 4.36 ± 0.51 (3%, 1,000,000) Deacetylated XG group 3 4.01 ± 0.36 (5%, 1,000,000) Deacetylated XG group 4 4.22 ± 0.41 (1%, 5,000,000)

[0050] 2. Research on the Influence of the Above Deacetylated XG Joint Cavity Injection Preparations on the Lubricating Effect of Bovine Articular cartilage:

[0051] Bovine articular cartilage was peeled off by a special bovine bone steel knife and prepared into cartilage masses with uniform thickness (1.89 mm). The tribological indexes under lubrication of different injection preparations were tested on a reciprocating friction testing machine, including the following: dry friction group, NS group, SH group, deacetylated XG group 1 (Example 6, 1%, 1,000,000), deacetylated XG group 2 (Example 7, 3%, 1,000,000), deacetylated XG group 3 (Example 8, 5%, 1,000,000), deacetylated XG group 4 (Example 9, 1%, 5,000,000), ordinary XG group (1%, 1,000,000), and SH control group. Parameters of the testing machine were set as follows: test force: 3 N, and reciprocating frequency: 1 Hz. Test results were shown in Table 2 below.

TABLE-US-00007 TABLE 2 Influence on the lubricating effect of bovine articular cartilage Group Pathological score Dry friction group 0.342 ± 0.09 NS group 0.151 ± 0.08 SH group 0.113 ± 0.06 Ordinary XG group 0.081 ± 0.05 (1%, 1,000,000) Deacetylated XG group 1 0.083 ± 0.07 (1%, 1,000,000) Deacetylated XG group 2 0.073 ± 0.06 (3%, 1,000,000) Deacetylated XG group 3 0.064 ± 0.08 (5%, 1,000,000) Deacetylated XG group 4 0.071 ± 0.10 (1%, 5,000,000)

[0052] It can be seen from the results that, on the reciprocating friction testing machine, the bovine knee articular cartilage showed lubricating effects under the lubrication of deacetylated XG and ordinary XG better than that under the lubrication of SH.

[0053] 3. Research on the Biocompatibility of the Above Deacetylated XG Joint Cavity Injection Preparations:

[0054] 80 SD rats were randomly divided into 8 groups, each with 10 rats (half male and half female): negative control group (NS), deacetylated XG group 1 (Example 6, 1%, 1,000,000), deacetylated XG group 2 (Example 7, 3%, 1,000,000), deacetylated XG group 3 (Example 8, 5%, 1,000,000), deacetylated XG group 4 (Example 9, 1%, 5,000,000), deacetylated XG group 5 (Example 10, 5%, 5,000,000), ordinary XG group 1 (5%, 1,000,000), and ordinary XG group 2 (5%, 5,000,000). The joint cavities were injected with the corresponding drugs or NS once every 2 weeks at a dosage of 0.06 mL/joint, and the administration lasted for 6 months. Two weeks after the last administration, animals were anesthetized and blood was collected for blood routine examination; and animals were sacrificed, spleens were collected and weighed to calculate an organ coefficient (%).

[0055] Results showed that the injection of ordinary XG caused a significant increase in spleen weight and monocyte proportion, but the repeated injection of high-dosage deacetylated XG did not cause adverse reactions of organs and tissues and abnormal results of blood routine examination.

[0056] The above experiments show that the long-term repeated injection of high-dosage XG caused adverse reactions such as increase in spleen weight and monocyte proportion, but the long-term repeated injection of high-dosage deacetylated XG caused no adverse reactions, which indicates that the deacetylated XG has a sate dosage range for OA treatment that is significantly wider than that of ordinary XG, and also has a significant advantage in biocompatibility.

TABLE-US-00008 TABLE 3 Influence of deacetylated XG on monocytes Monocyte Monocyte Group number proportion NS group 0.30 ± 0.04 3.20 ± 0.73 Ordinary XG group 0.38 ± 0.11 3.85 ± 0.88 (5%, 1,000,000) Ordinary XG group 0.67 ± 0.15 5.25 ± 1.24 (5%, 5,000,000) Deacetylated XG group 1 0.31 ± 0.03 3.18 ± 0.69 (1%, 1,000,000) Deacetylated XG group 2 0.29 ± 0.05 3.17 ± 0.78 (3%, 1,000,000) Deacetylated XG group 3 0.32 ± 0.03 3.31 ± 0.65 (5%, 1,000,000) Deacetylated XG group 4 0.34 ± 0.05 3.51 ± 0.77 (1%, 5,000,000) Deacetylated XG group 5 0.35 ± 0.09 3.69 ± 0.87 (5%, 5,000,000)

TABLE-US-00009 TABLE 4 Influence of deacetylated XG on spleen weight Spleen weight Organ coefficient Group (g) (%) NS group 1.301 ± 0.12 0.26 ± 0.05 Ordinary XG group 1.339 ± 0.11 0.28 ± 0.04 (5%, 1,000,000) Ordinary XG group 1.591 ± 0.15 0.35 ± 0.04 (5%, 5,000,000) Deacetylated XG group 1 1.283 ± 0.17 0.25 ± 0.05 (1%, 1,000,000) Deacetylated XG group 2 1.302 ± 0.23 0.26 ± 0.06 (3%, 1,000,000) Deacetylated XG group 3 1.300 ± 0.09 0.25 ± 0.03 (5%, 1,000,000) Deacetylated XG group 4 1.298 ± 0.08 0.26 ± 0.04 (1%, 5,000,000) Deacetylated XG group 5 1.314 ± 0.13 0.27 ± 0.04 (5%, 5,000,000)

[0057] The above implementations should not he considered as a limitation on the protection scope of the present disclosure. It should be appreciated by those skilled in the art that any alternative improvement or change made to the implementations of the present disclosure falls within the protection scope of the present disclosure.

[0058] Anything not described in detail in the present disclosure may be a widely-known technology for those skilled in the art.