Oligosaccharide compound preparation capable of remarkably relieving irinotecan chemotherapy-induced intestinal injury

Abstract

The invention discloses an oligosaccharide compound preparation capable of remarkably relieving irinotecan chemotherapy-induced intestinal injury, which is prepared by compounding alginic acid oligosaccharide, fructo-oligosaccharide and stachyose according to the mass ratio of (40-60): (20-30): (20-30), the alginic acid oligosaccharide, the fructo-oligosaccharide and the stachyose are high in safety and free of side effects, and can be applied to relieving irinotecan-induced chemotherapy-induced intestinal injury, and the three components cooperate to effectively promote proliferation of beneficial bacteria, so that the curative effect of irinotecan on the chemotherapy-induced intestinal injury is improved. The micro-ecological environment of the intestinal tract is improved from the source; intestinal inflammation caused by chemotherapy is rapidly inhibited, and release of inflammatory factors is reduced; the damaged intestinal mucosa is repaired, and the intestinal barrier function is enhanced; the technical effect of the irinotecan preparation is obviously better than that of a single oligosaccharide preparation or a pairwise compounded preparation, and the irinotecan preparation can effectively solve the clinical pain point of intestinal discomfort in the irinotecan chemotherapy process, so that smooth chemotherapy is guaranteed. Meanwhile, the preparation process disclosed by the invention does not need complex equipment, is simple and convenient to operate, has strong controllability, is suitable for industrial large-scale production, and has a good industrial prospect.

Description

Technical Field

[0001] This invention belongs to the field of functional food and health food technology, specifically relating to an oligosaccharide compound preparation that can significantly reduce irinotecan chemotherapy-induced intestinal damage. Background Technology

[0002] Irinotecan (CPT-11) is a widely used chemotherapy drug for various solid tumors, including colorectal cancer. However, its severe dose-limiting toxicity leads to intestinal damage, significantly impacting patients' quality of life and often causing chemotherapy regimen interruptions, thus affecting treatment efficacy. The main mechanism by which CPT-11 causes intestinal damage is the accumulation of its active metabolite SN-38 in the intestine, directly damaging intestinal epithelial cells and causing dysbiosis, thereby triggering severe intestinal inflammation, apoptosis, and disruption of the intestinal barrier function. Currently, treatment for CPT-11 colitis is mostly symptomatic and supportive, such as using antidiarrheal drugs like loperamide, but the effects are limited and may lead to constipation.

[0003] Alginate oligosaccharides (AOS) are naturally found in the cell walls of brown algae such as kelp, nori, and wakame. They are small-molecule oligosaccharides obtained by degrading large-molecule alginic acid through enzymatic and acid hydrolysis. Fructooligosaccharides (FOS) are naturally found in plants such as Jerusalem artichoke, chicory, banana, onion, and garlic. They are mostly prepared from Jerusalem artichoke / chicory through enzymatic hydrolysis or extraction and purification. Stachyose (SS) are naturally found in various plants and are prepared through plant extraction and purification. Alginate oligosaccharides, fructooligosaccharides, and stachyose (SS) all have the function of promoting the proliferation of beneficial bacteria such as Bifidobacterium and Lactobacillus, and inhibiting the growth of harmful bacteria such as Clostridium perfringens and Escherichia coli. However, to date, there are no reports of combining alginate oligosaccharides, fructooligosaccharides, and stachyose to significantly reduce irinotecan-induced intestinal injury. Summary of the Invention

[0004] The present invention aims to solve the above-mentioned technical problems existing in the prior art and provide an oligosaccharide compound preparation that can significantly reduce irinotecan chemotherapy-induced intestinal damage.

[0005] The technical solution of the present invention is: an oligosaccharide compound preparation that can significantly reduce chemotherapy-induced intestinal damage caused by irinotecan, composed of alginate oligosaccharide, fructooligosaccharide and stachyose, wherein the mass ratio of alginate oligosaccharide, fructooligosaccharide and stachyose is 40-60:20-30:20-30.

[0006] The preferred mass ratio of the alginate oligosaccharide, fructooligosaccharide, and stachyose is 50:25:25.

[0007] Preferably, the total mass of the alginate oligosaccharide, fructooligosaccharide and stachyose is 20% of the total mass, with the remainder being maltodextrin.

[0008] Preferably, the alginate oligosaccharide has a degree of polymerization of 2-20, wherein the ratio of guluronic acid to mannuronic acid is 1:1, and it is prepared by enzymatic method with a purity ≥90%; the fructooligosaccharide has a degree of polymerization of 2-8, is extracted from chicory root, and has a purity ≥85%; the stachyose is an oligotetraglucose composed of galactobiose, glucose, and fructose with a purity ≥80%.

[0009] This invention is a compound of alginate oligosaccharides, fructooligosaccharides, and stachyose, which are highly safe and have no side effects. It can be used to alleviate chemotherapy-induced intestinal damage caused by irinotecan. The three compounds synergistically promote the proliferation of beneficial bacteria, fundamentally improving the intestinal microecological environment; rapidly inhibit chemotherapy-induced intestinal inflammation, reducing the release of inflammatory factors; repair damaged intestinal mucosa, and enhance intestinal barrier function. The resulting technical effects are significantly superior to those of single oligosaccharides or formulations composed of two compounds, i.e., a quantitative change occurs, effectively addressing the clinical pain point of intestinal discomfort during irinotecan chemotherapy, thereby ensuring the smooth progress of chemotherapy. Furthermore, the preparation process of this invention requires no complex equipment, is simple to operate, highly controllable, suitable for large-scale industrial production, and has good industrialization prospects. Attached Figure Description

[0010] Figure 1 This is a pathological section of mouse jejunal tissue structure and its quantitative diagram, as described in an embodiment of the present invention. Detailed Implementation

[0011] This invention discloses an oligosaccharide compound preparation that can significantly alleviate chemotherapy-induced intestinal damage caused by irinotecan. It comprises alginate oligosaccharide, fructooligosaccharide, stachyose, and maltodextrin, with the following weight proportions: alginate oligosaccharide 50 parts, fructooligosaccharide 25 parts, stachyose 25 parts, and maltodextrin 400 parts. The alginate oligosaccharide has a degree of polymerization of 2-20, wherein the ratio of guluronic acid to mannuronic acid is 1:1, and it is prepared by enzymatic method with a purity ≥90%. The fructooligosaccharide has a degree of polymerization of 2-8, is extracted from chicory root, and has a purity ≥85%. The stachyose is a tetraoligosaccharide composed of galactobiose, glucose, and fructose, with a purity ≥80%.

[0012] Prepare according to the following steps: 1. Raw material pretreatment: Alginate oligosaccharides, fructooligosaccharides, and stachyose are passed through an 80-mesh sieve to remove impurities; 2. Mixing: According to the formula ratio, first add the alginate oligosaccharide to the three-dimensional mixer and stir for 10 minutes; then add the fructooligosaccharide and continue stirring for 15 minutes; finally add the stachyose and maltodextrin and stir for 20 minutes to ensure that the coefficient of variation of the uniformity of the mixture is ≤5%.

[0013] 3. Forming and Packaging: The uniformly mixed materials are packaged using an automatic packaging machine, with each bag containing 10g net weight. The packaging material is a barrier composite film (PET / AL / PE) to prevent moisture absorption and oxidation. After packaging, a sealing test is performed to ensure compliance.

[0014] Experiment: Pharmacodynamic evaluation of embodiments of the present invention (animal experiment).

[0015] 1. Experimental animals and grouping The mice were housed in a controlled environment (room temperature 22±2 ℃; humidity 55±5 %) with free access to food and water. Healthy and active ICR mice were selected for acclimatization for 3 days, during which they were fed a basal diet twice a day. After the feeding period, 72 healthy and active ICR mice with an initial weight of 28 ± 2.08 g were selected and randomly divided into 9 groups (CK control group, CPT model group, alginate oligosaccharide (AOS) single treatment group (A), fructooligosaccharide (FOS) single treatment group (F), stachyose (SS) single treatment group (S), AOS+FOS co-treatment group (A+F), AOS+SS co-treatment group (A+S), FOS+SS co-treatment group (F+S), and AOS+FOS+SS multi-treatment group (A+F+S, i.e., the groups of the present invention), with 8 mice in each group. The dosage of each single treatment group was 300 mg / kg, and the dosage of each co-treatment group was: AOS (150 mg / kg) + FOS (150 mg / kg) group, AOS (150 mg / kg) + SS (150 mg / kg) group, FOS (150 mg / kg) + SS (150 mg / kg) group, and FOS (150 mg / kg) + SS (150 mg / kg) group. The treatment groups were divided into two groups: AOS (100 mg / kg) + FOS (100 mg / kg) + SS (100 mg / kg). The experiment lasted 24 days. From day 1 to day 16, the CK and CPT groups were administered saline by gavage daily, while the seven oligosaccharide treatment groups were administered solutions containing the corresponding oligosaccharide preparations by gavage at 7:30 and 17:00 daily, with each mouse receiving 0.2 ml of the solution. From day 17 to day 24, mice in the CPT group and each oligosaccharide treatment group were injected intraperitoneally with 60 mg / kg CPT-11 solution at 11:00 daily.

[0016] Observe the health status and feeding of the mice daily, and record their weight changes and fecal condition. Change the mouse bedding every 3 days and the mice's drinking water every 2 days. Keep the animal room dry and well-ventilated, and alternate between light and dark for 12 hours a day.

[0017] 2. Sample collection and calculation formula At the end of the feeding experiment, the mice were fasted for 12 hours, weighed, and all mice in each group were euthanized. Serum and tissue samples were collected and stored at -80°C.

[0018] Change in body weight = W t - W0 In the formula: W0 is the mouse body weight (g) at the start of CPT-11 injection; W t The weight (g) of the mice at the end of the experiment.

[0019] Disease Activity Index Evaluation Criteria: Table 1 Disease Activity Index

[0020] Note: Stool characteristics (± indicates loose stool with less water content, + indicates loose stool with more water content); blood in stool (± indicates faint blood in stool, + indicates visible bloodstains, ++ indicates a large amount of blood, >++ indicates a large amount of blood in stool).

[0021] Jejunal histopathological scoring criteria: Table 2. Jejunal tissue pathology scores

[0022] 3. Experimental Results and Analysis 3.1 To evaluate the protective effect of the compound oligosaccharide preparation described in this invention against irinotecan-induced intestinal pathological damage, the results of the changes in pathological indicators of mice in each group in the in vivo animal experiment are shown in Table 3.

[0023] Table 3. Changes in pathological indicators of mice in each group

[0024] Note: A represents alginate oligosaccharide, F represents fructooligosaccharide, and S represents stachyose. + indicates increased mouse weight, and - indicates decreased mouse weight.

[0025] N=8, compared to the CK group, # p <0.05, ## p <0.01; compared with the CPT group, p <0.05, p <0.01.

[0026] The results showed that after irinotecan injection, the disease index of mice in the model group increased significantly, while the oligosaccharides... After treatment and intervention, the disease activity index was effectively improved, with the oligosaccharide multitreatment showing particularly significant improvement. After irinotecan injury and intervention with various oligosaccharide treatments, the weight loss of mice in each group was compared according to the formula for calculating weight loss. Oligosaccharide multitreatment significantly reduced the weight loss of mice. After irinotecan injury, the length of the colon in mice was shortened, and intervention with various oligosaccharide treatments could slow down this trend. Statistical analysis of the colon length of mice in each group showed that oligosaccharide multitreatment could significantly reverse the shortening of the colon length in mice.

[0027] 3.2 The occurrence and development of intestinal mucosal injury are often closely related to the abnormal activation of inflammatory responses. The expression levels of pro-inflammatory cytokines TNF-α and IL-6 are important indicators reflecting the degree of intestinal inflammation and the state of mucosal damage. To clarify the intervention effect of the compound oligosaccharide preparation described in this invention on irinotecan-induced intestinal inflammatory response, the levels of TNF-α and IL-6 in the serum of mice in each group were detected using an ELISA kit. The experimental results are shown in Table 4.

[0028] Table 4. Changes in inflammatory factor indices in each group of mice

[0029] Note: A represents alginate oligosaccharide, F represents fructooligosaccharide, and S represents stachyose. + indicates increased mouse body weight, and - indicates decreased mouse body weight. N=8, compared with the CK group, # p <0.05, ## p <0.01; compared with the CPT group, p <0.05, p <0.01.

[0030] The results showed that, compared with the normal control group, the serum levels of pro-inflammatory cytokines IL-6 and TNF-α in mice in the irinotecan model group were significantly increased, indicating that irinotecan-induced intestinal mucosal damage was accompanied by significant inflammatory response activation. After intervention with various oligosaccharide treatment groups, the elevated levels of IL-6 and TNF-α induced by irinotecan were effectively downregulated, with multiple oligosaccharide treatments showing the most significant inhibitory effect on both pro-inflammatory cytokines. These results indicate that the compound oligosaccharide preparation described in this invention can more significantly inhibit irinotecan-induced inflammatory responses, reduce the release levels of pro-inflammatory cytokines, and has a more significant protective effect against intestinal mucosal damage.

[0031] 3.3 To clarify the protective effect of the compound oligosaccharide preparation described in this invention against irinotecan-induced jejunal tissue structural damage, intestinal tissues from mice in the CK group, CPT group, and the groups with the best protective effects (A, A+F, and A+F+S) were selected for pathological section preparation. HE-stained sections were used for pathological analysis. The experimental results are shown in Figure 1. Figure 1 Compared with the CK group, # p <0.05, ## p <0.01; compared with the CPT group, p <0.05, p <0.01. The results showed that compared with the normal control group, the irinotecan model group mice exhibited significant pathological damage to the intestinal epithelium and mucosa, specifically manifested as jejunal crypt destruction, cell vacuolation, shortened jejunal villi, and characteristic pathological changes such as inflammatory cell infiltration. After oligosaccharide multitreatment intervention, the irinotecan-induced jejunal tissue structural abnormalities were significantly improved. Analysis of HE sections of mice from each group based on jejunal tissue pathological scores showed that irinotecan treatment significantly increased the histopathological score, while oligosaccharide multitreatment intervention significantly decreased the score. Furthermore, the villi length to crypt depth ratio is an important indicator of jejunal digestive capacity. Measurements of small intestinal villi length and crypt depth further confirmed that oligosaccharide multitreatment intervention effectively reversed the decrease in the villi length to crypt depth ratio caused by irinotecan. These experimental results fully demonstrate that the compound oligosaccharide preparation described in this invention can significantly improve irinotecan-induced intestinal tissue structural damage and has a better protective effect against irinotecan-induced jejunal tissue pathological damage.

Claims

1. An oligosaccharide compound preparation that can significantly reduce irinotecan-induced chemotherapy-induced intestinal damage, characterized in that... It is composed of alginate oligosaccharides, fructooligosaccharides and stachyose, wherein the mass ratio of alginate oligosaccharides, fructooligosaccharides and stachyose is 40-60:20-30:20-30.

2. The oligosaccharide compound preparation according to claim 1, which can significantly reduce irinotecan-induced chemotherapy-induced intestinal damage, is characterized in that: The mass ratio of alginate oligosaccharide, fructooligosaccharide, and stachyose is 50:25:

25.

3. The oligosaccharide compound preparation according to claim 1 or 2, which can significantly reduce irinotecan-induced chemotherapy-induced intestinal damage, is characterized in that: The alginate oligosaccharide, fructooligosaccharide and stachyose together account for 20% of the total mass, with the remainder being maltodextrin.

4. The oligosaccharide compound preparation according to claim 3, which can significantly reduce irinotecan-induced chemotherapy-induced intestinal damage, is characterized in that: The alginate oligosaccharide has a degree of polymerization of 2-20, wherein the ratio of guluronic acid to mannuronic acid is 1:1, and it is prepared by enzymatic method with a purity of ≥90%; the fructooligosaccharide has a degree of polymerization of 2-8, is extracted from chicory root, and has a purity of ≥85%; the stachyose is an oligotetrasaccharide composed of galactobiose, glucose, and fructose with a purity of ≥80%.

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