CN116327811A

CN116327811A - Composite prebiotics and composite probiotics combined for treating allergic rhinitis

Info

Publication number: CN116327811A
Application number: CN202211358485.2A
Authority: CN
Inventors: 洪昱文
Original assignee: Biogrowing Shanghai Co ltd; Yongzetai Biotechnology Beijing Co ltd
Current assignee: Biogrowing Shanghai Co ltd; Yongzetai Biotechnology Beijing Co ltd
Priority date: 2022-11-01
Filing date: 2022-11-01
Publication date: 2023-06-27
Also published as: WO2024093938A1

Abstract

The invention provides a probiotic preparation and prebiotic preparation combined drug for treating allergic rhinitis, in particular to a probiotic preparation which comprises lactobacillus acidophilus, lactobacillus paracasei, lactobacillus grignard, lactobacillus longum, bifidobacterium lactis and bifidobacterium infantis; the prebiotic formulation comprises fructo-oligosaccharides and xylo-oligosaccharides. The probiotic preparation and the prebiotic preparation can cooperate to enhance the effect of treating allergic rhinitis.

Description

Composite prebiotics and composite probiotics combined for treating allergic rhinitis

Technical Field

The invention relates to the field of biological medicine, in particular to a method for treating allergic rhinitis by combining composite prebiotics and composite probiotics.

Background

Allergic rhinitis is a common allergic disease of the respiratory tract in the otorhinolaryngology, and in recent years, related studies have confirmed that allergic rhinitis is type I allergic reaction mediated by IgE after exposure of the nasal mucosa to allergens. Clinical trials prove that allergic rhinitis is caused by multi-factor induction, including environment, genes and the like, genetic factors, allergens and the like, and can cause the disease.

In the progress of the disease, various cytokines, immunocompetent cells and the like are involved, clinical symptoms are typical, mainly runny nose, itching nose and sneeze are the main symptoms, and also the symptom of hyposmia occurs in some patients. Allergic rhinitis is a common disease, often combined with adenoid hypertrophy, asthma, conjunctivitis and the like, and the interaction aggravates the illness and also affects the work and study of patients. Therefore, the medicine should be used for treating in time and relieving symptom reaction. Antihistamines are more commonly used in conventional treatments and can better improve clinical symptoms, but must be taken for a long period of time.

Along with the progress of clinical researches in recent years, the prebiotics and probiotics have good regulation effect on the immune function of patients with allergic rhinitis and positive effect on improving the prognosis of the patients. Clinical trials of using prebiotics, probiotics to treat allergic rhinitis have shown a certain advantage.

Disclosure of Invention

The invention aims to provide a composite preparation containing a probiotic preparation and a prebiotic preparation, which is used for preventing and/or treating allergic rhinitis.

In a first aspect of the present invention, there is provided a composite formulation comprising:

a prebiotic formulation comprising fructo-oligosaccharides, xylo-oligosaccharides; and

the probiotic preparation is a live bacterial preparation and comprises lactobacillus acidophilus, lactobacillus paracasei, lactobacillus gasseri, bifidobacterium longum, bifidobacterium lactis and bifidobacterium infantis;

wherein, lactobacillus acidophilus is CCTCC NO: lactobacillus acidophilus of M2013337;

the lactobacillus paracasei is a lactobacillus paracasei with a preservation number of CCTCC NO: m2013691 Lactobacillus paracasei

The Lactobacillus gasseri has a preservation number of CCTCC NO: lactobacillus gasseri of M2019829;

the bifidobacterium longum is a strain with a preservation number of CCTCC NO: m2013689 lactobacillus bifidus;

the bifidobacterium lactis is a compound having a preservation number of CCTCC NO: bifidobacterium lactis of M2013336;

the bifidobacterium infantis is a strain with a preservation number of CCTCC NO: bifidobacterium infantis of M2013688.

In another preferred embodiment, the weight ratio of the probiotic preparation to the prebiotic preparation is 1:5-20, preferably 1:5-10.

In another preferred example, the prebiotic formulation comprises the following ingredients in parts by weight:

20-70 parts by weight, preferably 30-60 parts by weight, more preferably 40-60 parts by weight of fructooligosaccharides;

1 to 20 parts by weight, preferably 2 to 10 parts by weight, more preferably 2 to 5 parts by weight of xylo-oligosaccharide.

In another preferred embodiment, the prebiotic formulation comprises the following ingredients, based on total weight of the prebiotic formulation:

20-70wt%, preferably 30-60wt%, more preferably 40-60wt%, e.g. 45wt%, 50 wt wt%, 55wt% of fructooligosaccharides;

the xylo-oligosaccharide is 1-20wt%, preferably 2-10wt%, more preferably 2-5wt%, such as 2wt%, 5wt%, 8wt%.

In another preferred embodiment, the prebiotic formulation further comprises inulin and/or galactooligosaccharides.

In another preferred embodiment, the prebiotic formulation further optionally comprises one or more of isomaltose, soy oligosaccharides, lactulose, mannooligosaccharides, chitosan oligosaccharides, dextran, coriolus versicolor polysaccharides, xylitol, mannitol, lactitol.

in another preferred embodiment, the degree of polymerization of fructose in the fructo-oligosaccharides is 2-20, preferably 2-10, more preferably 2-7.

In another preferred embodiment, the fructooligosaccharides have a pH of 4.5 to 7.0, preferably 6.0 to 7.0, more preferably 6.0 to 6.8.

In another preferred embodiment, the degree of polymerization of xylose in the xylooligosaccharide is 2-10, preferably 2-7.

In another preferred embodiment, the pH of the xylooligosaccharide is from 3.5 to 6.5, preferably from 3.5 to 5.0, more preferably from 3.5 to 4.0.

In another preferred embodiment, the polymerization degree of fructose in the inulin is 2 to 60, preferably 10 to 50, more preferably 20 to 40.

In another preferred embodiment, the galacto-oligosaccharide comprises the following components on a dry weight basis:

90-100 wt% galactose, preferably 92-100 wt%;

lactose 0-10wt%, preferably 0-7wt%, more preferably 0-5wt%;

glucose 0-3wt%, preferably 0-2wt%, more preferably 0-1wt%.

In another preferred embodiment, the degree of polymerization of the galacto-oligosaccharides is 2-6, preferably 2-4.

In another preferred embodiment, the pH of the galacto-oligosaccharide is 3.5-6.5, preferably 4.0-6.0, more preferably 5.0.

In another preferred embodiment, the preparation method of the prebiotic formulation comprises the following steps:

(1) Weighing xylooligosaccharide or optionally inulin, and stirring and mixing to obtain mixed powder;

(2) And (3) stirring and mixing the mixed powder obtained in the step (1) with fructooligosaccharide powder and optionally galactooligosaccharide powder to obtain the prebiotic preparation.

In another preferred embodiment, the xylo-oligosaccharide, inulin, galacto-oligosaccharide powder and fructo-oligosaccharide powder are optionally pre-treated before mixing, wherein the pre-treatment is sieving.

In another preferred embodiment, the sieving refers to passing through a 40 mesh screen.

In another preferred embodiment, the stirring speed is 500-1000rpm/min.

In another preferred embodiment, in the step (1), the mixture is stirred and mixed for 5 to 10 minutes.

In another preferred embodiment, in the step (2), the mixture is stirred and mixed for 15 to 30 minutes.

In another preferred example, the probiotic preparation comprises the following ingredients in parts by number of viable bacteria:

in another preferred embodiment, the probiotic preparation comprises the following ingredients in terms of total viable count of the preparation:

in another preferred embodiment, the population of bacteria in the probiotic preparation is viable bacteria.

In another preferred embodiment, the probiotic preparation is mixed gradient by equal incremental method.

In another preferred embodiment, the probiotic preparation has a viable count of lactobacillus of (0.1-20) ×10 ¹⁰ CFU/g, preferably (1-10). Times.10 ¹⁰ CFU/g, preferably > 2.0X10 ¹⁰ CFU/g。

In another preferred example, the probiotic preparation further comprises fructo-oligosaccharide, erythritol, stachyose, acerola cherry concentrated powder and red pomegranate fruit juice powder.

In another preferred embodiment, the complex formulation is prepared by gradient mixing in equal increments.

In another preferred example, the composite preparation is prepared by mixing raw materials of the probiotic preparation and raw materials of the probiotic preparation in an equal-amount incremental method gradient.

In another preferred embodiment, the complex formulation is prepared by mixing a probiotic formulation and a prebiotic formulation.

In another preferred embodiment, the complex formulation comprises a probiotic formulation and a prebiotic formulation, packaged separately.

In another preferred example, the dosage form of the composite preparation is powder, capsule, suspension, emulsion, syrup, liquid spray, ointment or gel.

In another preferred embodiment, the dosage form of the composite formulation is a solid dosage form, a semi-solid dosage form, or a liquid dosage form.

In another preferred embodiment, the complex formulation is administered orally.

In another preferred embodiment, the composite formulation further comprises an adjuvant selected from the group consisting of: flavors, sweeteners, effervescent agents, dispersants, suspending agents, emulsifying agents, preservatives, or combinations thereof.

In another preferred embodiment, the probiotic preparation, the formulation of the prebiotic preparation are each independently a solid, semi-solid or liquid formulation.

In another preferred example, the formulation of the probiotic preparation and the prebiotic preparation are each independently powder, capsule, solution, suspension, emulsion, liquid spray, ointment and gel.

In another preferred embodiment, the probiotic preparation and the prebiotic preparation are each independently administered orally.

In another preferred embodiment, the probiotic preparation and the prebiotic preparation may be administered simultaneously, separately or sequentially, preferably separately.

In a second aspect of the present invention there is provided the use of a complex formulation as described in the first aspect of the present invention for the preparation of a composition for the prevention and/or treatment of allergic rhinitis.

In another preferred example, the allergic rhinitis includes hereditary allergic rhinitis and allergen-exposed allergic rhinitis.

In another preferred example, the allergic rhinitis includes perennial allergic rhinitis and seasonal allergic rhinitis.

In another preferred embodiment, the prevention and/or treatment of allergic rhinitis comprises one or more features selected from the group consisting of:

(1) The TNSS score of the patient is reduced, preferably by 1.0 point or more within 91 days, preferably by 2.0 points or more within 91 days, more preferably by 3.0 points or more within 91 days;

(2) The intestinal flora diversity is improved.

In a third aspect of the invention there is provided a pharmaceutical composition comprising a complex formulation according to the first aspect of the invention; and a pharmaceutically acceptable carrier.

In a fourth aspect of the invention, there is provided a product combination comprising a first package enclosing a prebiotic formulation and a second package enclosing a probiotic formulation.

In another preferred embodiment, the probiotic preparation is as described in the first aspect of the invention.

In another preferred embodiment, the prebiotic formulation is as described in the first aspect of the invention.

In another preferred embodiment, the product combination optionally comprises instructions for use.

In another preferred embodiment, the instructions for use state that the probiotic preparation and the prebiotic preparation are combined, thereby preventing and/or treating allergic rhinitis.

In another preferred embodiment, the probiotic preparation and the prebiotic preparation are administered simultaneously in the prevention and/or treatment of allergic rhinitis.

In another preferred embodiment, the probiotic preparation and the prebiotic preparation are each individually packaged.

In another preferred embodiment, the probiotic preparation and the prebiotic preparation are mixed.

In a fifth aspect of the present invention there is provided a method of preventing and/or treating allergic rhinitis, the method comprising the steps of: a composite formulation according to the first aspect of the invention is administered to a subject in need thereof.

In another preferred embodiment, the subject is a mammal, preferably a human.

It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (e.g., in the examples) may be combined with each other to constitute new or preferred technical solutions. And are limited to a space, and are not described in detail herein.

Drawings

Figure 1 shows the process for preparing a prebiotic solid powder.

Figure 2 shows the preparation of probiotic solid powders.

Figure 3 shows a diversity differences in α between different times of treatment groups compared to baseline phase: 31 days (FIG. 3A); 61 days (fig. 3B); 91 days (FIG. 3C).

Figure 4 shows the alpha diversity differences between the treated and control groups: baseline period (fig. 4A); 31 days (FIG. 4B); 61 days (fig. 4C); 91 days (FIG. 4D).

Detailed Description

The present inventors have made extensive and intensive studies to unexpectedly develop a composition comprising probiotics for the first timeThe invention adopts a combination of a probiotic preparation and a specific prebiotic preparation which are scientifically verified, on one hand, the antiallergic effect of the raw materials is ensured to be exerted, and on the other hand, the prebiotic is used as a nutrient substance of the probiotics, so that the proliferation of the edible probiotics in intestinal tracts is supported, and the raw materials are provided for the fermentation of dietary fibers by the probiotics to produce short-chain fatty acids. The invention fully considers the probiotics and the prebiotics to be combined to exert' 1+1>2', not only ensures the dosage effect of the prebiotics, but also supplements the sufficient viable count (more than or equal to 2.0X10) of the probiotics ¹⁰ CFU/bar). The present invention has been completed on the basis of this finding.

Terminology

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the terms "comprising," "including," and "containing" are used interchangeably, and include not only closed-form definitions, but also semi-closed-form and open-form definitions. In other words, the term includes "consisting of … …", "consisting essentially of … …".

Composite preparation

The invention provides a composite preparation comprising a prebiotic preparation and a probiotic preparation.

The compound preparation adopts the combination of the probiotics and the prebiotics which are scientifically verified, on one hand, the antiallergic effect of the raw materials is ensured to be exerted, on the other hand, the prebiotics are used as the nutrient substances of the probiotics, so that the proliferation of the edible probiotics in the intestinal tract is supported, and the raw materials are provided for the fermentation of dietary fibers by the probiotics to produce short-chain fatty acids.

The compound preparation enables the combination of the probiotics and the prebiotics to exert' 1+1>2', not only ensures the dosage effect of the prebiotics, but also supplements the sufficient viable count (more than or equal to 2.0X10) of the probiotics ¹⁰ CFU/bar).

The invention plays a complementary effect through the combination of the composite prebiotics and the composite probiotics, regulates intestinal bacteria, eliminates inflammation, improves metabolism and balances immunity; the prebiotics and probiotics used are supported by a large number of animal experiments and clinical experiments; fully considering the dose effect exerted by the prebiotic effect; the total viable count is high, and the used strains are reasonably proportioned. Radically relieving allergic symptoms.

The probiotic preparation is a powder mixture, and is prepared by mixing 6 raw materials such as probiotic powder, fructo-oligosaccharide, erythritol, stachyose, pomegranate fruit powder, acerola cherry powder and the like in a gradient manner by an equal-amount increasing method, and has granularity of 60-100 meshes, and the mixed probiotic preparation is light red to light pink fluffy powder, sweet taste and no peculiar smell.

The prebiotic preparation comprises xylo-oligosaccharide and fructo-oligosaccharide, and the two prebiotics have synergistic colonization effects on the probiotic preparation, and particularly have excellent effects on the colonization of bifidobacteria.

Compositions and applications

The compositions of the present invention include (but are not limited to): pharmaceutical compositions, food compositions, health care compositions, dietary supplements, and the like.

Representatively, the complex formulation of the present invention may be formulated into a nutraceutical composition such as a dosage form of a tablet, capsule, powder, granule, solution, lozenge, jelly, suspension, or the like. The nutraceutical compositions can be prepared by commonly known preparation techniques and suitable nutraceutical additives can be added to the nutraceuticals.

The compositions of the present invention may also include a pharmaceutically, food, nutraceutical or dietary acceptable carrier. "pharmaceutically, food, nutraceutical or dietary acceptable carrier" means: one or more compatible solid or liquid filler or gel materials which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. "compatible" as used herein means that the components of the composition are capable of blending with and between the compounds of the present invention without significantly reducing the efficacy of the compounds. Pharmaceutically, food, health care or dietary acceptable carrier, examples of acceptable carrier parts are cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as hardFatty acids, magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyalcohol (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), and emulsifying agent (such as

) Wetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizing agents, antioxidants, preservatives, pyrogen-free water and the like.

The mode of administration of the compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, topical, preferably oral administration.

The formulation of the composition or the preparation provided by the invention is an oral preparation. Typically, solid dosage forms for oral administration or administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) Fillers or compatibilizers, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) Binders, for example, hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, e.g., glycerin; (d) Disintegrants, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent, such as paraffin; (f) an absorption accelerator, e.g., a quaternary amine compound; (g) Wetting agents, such as cetyl alcohol and glycerol monostearate; (h) an adsorbent, for example, kaolin; and (i) a lubricant, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other materials well known in the art. They may contain opacifying agents.

Liquid dosage forms for oral administration or administration include pharmaceutically acceptable emulsions, suspensions or tinctures. In addition to the active compound, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of these substances and the like.

In addition to these inert diluents, the compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active ingredient, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these substances, and the like.

The complex formulation of the present invention may be administered alone or in combination with other agents for the prevention and/or treatment of allergic rhinitis. Preferred co-administered drugs include those commonly used in the art for the treatment of allergic rhinitis, such as imidazoles, corticosteroids, antibiotics.

The composition is applied by applying a safe and effective amount of the composite preparation of the invention to human or non-human animals (such as rats, mice, dogs, cats, cows, chickens, ducks, monkeys, gorillas, etc.), wherein the application dose is an effective administration dose which can be accepted by pharmacy, food or health care products. As used herein, the term "safe and effective amount" refers to an amount that is functionally or actively produced by and acceptable to a human and/or animal. It will be appreciated by those of ordinary skill in the art that the "safe and effective amount" will vary with the form of the pharmaceutical composition, the route of administration, the adjuvant of the drug being used, the severity of the disease, and the combination with other drugs. For example, for a person weighing 60kg, the daily dosage is usually 0.1 to 1000g, preferably 1 to 600g, more preferably 2 to 300g. Of course, the particular dosage should also take into account factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled practitioner.

The main advantages of the invention include:

1. in the combination, the probiotic preparation and the prebiotic preparation can produce synergistic effect and enhance the effect of treating allergic rhinitis.

2. The two oligosaccharides in the prebiotics can produce a synergistic effect, and the effect of the colonization of probiotics (especially lactobacillus and bifidobacterium) is obviously improved.

The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. Percentages and parts are weight percentages and parts unless otherwise indicated.

Example 1 preparation of prebiotic solid powder

The formulation and composition of the prebiotic solid powder are shown in Table 1 below:

TABLE 1

The preparation method of the prebiotic solid powder is shown in figure 1, and specifically comprises the following steps:

1. sieving: sieving raw materials (galactooligosaccharide powder, fructooligosaccharide powder, xylooligosaccharide, inulin) with 40 mesh sieve (vibrating screen)

2. Weighing and proportioning:

(1) Weighing materials such as xylooligosaccharide, inulin and the like according to a formula, putting the materials into a GH500 type efficient three-dimensional motion mixer, setting the rotating speed to 600rpm/min, mixing for 5 minutes, and discharging to obtain mixed powder A;

(2) Putting the galactooligosaccharide powder, the fructooligosaccharide powder and the mixed powder A into a GH500 type efficient three-dimensional motion mixer, setting the rotating speed to 600rpm/min, mixing for 15 minutes, and discharging to obtain the mixed powder B.

3. Sub-packaging

The preparation process was carried out in a 100000-class clean zone.

The obtained prebiotic solid powder is white powder with uniform color, no foreign odor and no foreign impurity.

EXAMPLE 2 preparation of probiotic solid powder

The strain formulation of the probiotic solid powder is shown in table 2:

TABLE 2

Strain
	Lactobacillus acidophilus
Lactobacillus paracasei
	Lactobacillus gasseri
Lactobacillus longum
	Bifidobacterium lactis
Bifidobacterium infantis

The preparation method of the probiotic solid powder is shown in figure 2.

In addition, the probiotic solid powder also comprises ingredients such as fructo-oligosaccharide, erythritol, stachyose, acerola cherry concentrated powder, red pomegranate fruit juice powder and the like.

The probiotic preparation is live bacteria preparation, and the live bacteria number of lactobacillus is more than or equal to 2.0X10 when leaving factory ¹⁰ CFU/bar (200 billion/bar).

The probiotic preparation is fluffy powder, light red or light pink.

Example 3 synergistic effect verification

All strains were stored in medium at-80℃and glycerol was added as cryoprotectant at a volume fraction of 50%. For storage purposes, the strains (including lactobacillus and bifidobacterium) are anaerobically cultured at their optimal growth temperature (37 ℃). Subsequently, cells from the stock culture were subcultured (the seed strain was inoculated into the medium at a volume fraction of 1%) and further anaerobically cultured for 16 hours to form a seed culture. Subsequently, 0.32mL of seed culture was inoculated into test tubes, respectively, each containing 16mL of medium (medium containing 0.47g/mL of prebiotic as the sole carbon source for growth experiments at 37 ℃). Du Shixiao tubes were placed in each tube to test the generation of gas.

The grouping case includes: group a contains medium only (lactose); group B is a culture medium containing 0.47g/mL fructo-oligosaccharide; group C is a culture medium containing 0.47g/mL of xylo-oligosaccharide; group D is a culture medium containing 0.47g/mL of fructo-oligosaccharide and xylo-oligosaccharide, wherein the mass ratio of the fructo-oligosaccharide to the xylo-oligosaccharide is 5:1; group E is a culture medium containing 0.47g/mL of fructo-oligosaccharide and xylo-oligosaccharide, wherein the mass ratio of the fructo-oligosaccharide to the xylo-oligosaccharide is 9:1; group F is a culture medium containing 0.47g/mL of fructo-oligosaccharide and xylo-oligosaccharide, wherein the mass ratio of fructo-oligosaccharide to xylo-oligosaccharide is 11:1.

Cultures (16 mL) were anaerobically incubated at 37℃for 24h and 72h, and the Optical Density (OD) of the cultures was measured at 600nm wavelength using a spectrophotometer ₆₀₀ nm), pH was measured by a pH meter.

After the culture (16 mL) was anaerobically cultured at 37℃for 16 hours, bubble generation was observed in a Du Shixiao tube.

EXAMPLE 4 results of clinical Studies

This example is a prospective, randomized, double-blind, placebo-controlled, multi-center clinical trial.

Study population selection criteria: age 18-65 years (without boundary value), meets the relevant diagnosis standard in the Chinese allergic rhinitis diagnosis and treatment guidelines issued by the allergic rhinitis institute of the China medical society of 2018, has allergic rhinitis disease course of more than or equal to 1 year, has total allergic rhinitis symptom score (TNSS) of more than 4 minutes, has the requirement on treatment for patients, and does not use antihistamine drugs, corticosteroids or immunosuppressants within 1 month before screening.

Grouping and administration: patients meeting the conditions of the test group were randomly divided into control group (placebo, 60 cases), test group (combination of complex prebiotics and complex probiotics, 60 cases). The control group was given the complex prebiotic mimetic (15 g/bag, 2 bags/time, 2 times/day) +complex probiotic mimetic (2 g/bag, 2 bags/time, 2 times/day), the test group was given the complex prebiotic (15 g/bag, 2 bags/time, 2 times/day, each morning and evening, after being homogenized with 200-300mL warm water, taken once) +complex probiotic (2 g/bag, 2 bags/time, 2 times/day, each morning and evening with meals or after meals, and directly poured into the mouth for oral administration or warm water for oral administration after opening the package).

Biological sample index determination:

subjects were given stool and whole blood samples at baseline, 31 day (±3 day), 61 day (±3 day), 91 day (±3 day) follow-up after start of dosing.

Intestinal microbiota examination: fecal samples were collected and tested using 16SrDNA sequencing and qPCR.

Efficacy evaluation:

main therapeutic effect index

TNSS scoring was performed prior to treatment, at the end of treatment, during follow-up, and clinical efficacy of the test and control groups was compared by the difference between endpoint score and baseline score.

4.1TNSS score analysis

In subjects, TNSS score was different from baseline at day 31 visit, test group was-1.5+ -3.25, control group was-1.3+ -3.17; the difference between TNSS score and baseline at day 61 visit was-1.6+ -3.09 for the test group and-1.3+ -3.08 for the control group; the TNSS score was different from baseline on day 91, test group was-3.1.+ -. 3.82, and control group was-2.1.+ -. 2.60. The result shows that TNSS score index of the test group is reduced more than that of the control group, and the symptoms of allergic rhinitis are reduced. See table 3 below for details.

TABLE 3 variation of TNSS score from baseline at 31, 61, 91 day visit with drug

The result shows that the average value of TNSS symptom scores after 90 days of treatment is 4.4+/-3.24, the average value of score reduction is 3.1+/-3.82, the average value of symptom improvement rate is 35.524%, and the treatment significant effect proportion is as high as 33.3% (the symptom improvement rate is not less than 66%). The evaluation of each period shows that the preparation has longer acting process, the improvement effect gradually increases with the advancement of time, and the maximum time interval of the curative effect is 60-90 days.

Sensitivity analysis of TNSS scores was performed on subjects who were co-administered with imidazoles, corticosteroids, and antibiotics. The results are shown in table 4 below, which demonstrate that the complex formulation of the present invention still has an effect of alleviating symptoms of allergic rhinitis in subjects who are co-administered.

Table 4 changes in TNSS score from baseline at day 91 visit (sensitivity analysis)

4.2 analysis of intestinal flora influence

The effect on the intestinal flora is shown in table 5 below. It can be seen that the intestinal beneficial bacteria flora is obviously increased after the 31 st day of administration, and the intestinal beneficial bacteria flora is kept on the 61 st and 91 st days.

TABLE 5 changes in natural log values of intestinal flora from baseline at day 31, 61 and 91 visits with drug

Note that: taking the existence of a large number of 0 values into consideration, adding 1 to the original value, performing natural logarithmic transformation, and performing statistical analysis by adopting the transformed data.

EXAMPLE 5 clinical laboratory study results

The clinical samples and grouping method were the same as in example 4.

5.1 test methods

16s rDNA detection

Miseq high throughput sequencing was performed on an illumina Miseq sequencing platform followed by bioinformatic analysis.

The bioinformatics analysis includes the steps of:

i. sequencing data processing: and splicing the PE reads obtained by sequencing through flash software, controlling the quality, thus obtaining an effective sequence, correcting the sequence direction, and filtering the sequencing data to obtain a high-quality target sequence for subsequent analysis.

Out analysis and species annotation: and clustering the sequences by using Usearch software, clustering the sequences into OTUs according to 97% similarity, and removing chimeras in the clustering process to obtain the representative sequences of OUT. From this, 1 representative sequence was taken and analyzed for downstream species annotation using RDP classifier. The color shade defined by the magnitude W of the numerical value is represented by calculating the hemmap of the species annotation result of the software R construct. The high-abundance and low-abundance species are clustered in blocks, and the similarity and the difference of community compositions of a plurality of samples at each classification level are reflected through the color gradient and the similarity degree.

Alpha diversity analysis: the species abundance and diversity of the environmental community is estimated by a series of statistical analysis indexes, and the index for calculating the bacterial community diversity is expressed by Shannon index.

PCoA analysis (Principal Co-ordinates Analysis) was statistically analyzed and plotted by R software.

5.2 Effect of 16S rDNA analysis on alpha diversity of intestinal flora

5.2.1 alpha diversity differences at different times in treatment group:

as shown in fig. 3A, 3B, and 3C, there was no significant change in the alpha diversity of the flora at 31 days of treatment, and the alpha diversity of the flora increased significantly at 61 days and 91 days of treatment, as compared to the baseline period.

5.2.2 alpha diversity differences between treatment and control groups:

as shown in fig. 4A, 4B, 4C, and 4D, the treatment group (group X) and the control group (group Y) showed no significant difference in the alpha diversity at the baseline, and the treatment group showed a significant increase in the alpha diversity at 31 days, 61 days, and 91 days. (shannon index median: x_base=5.01, y_base=5.01; x_d31=4.86, y_d31=4.17; x_d61=5.25, y_d61=4.47; x_d91=5.36, y_d91=4.73).

Preservation of bacterial species

(1) Lactobacillus acidophilus LA-G80, with the preservation unit: china center for type culture Collection: the preservation number of the Chinese Wuhan university is: cctccc NO: m2013337, the preservation time is: 2013, 7, 16;

(2) Lactobacillus paracasei LPc-G110, deposit unit: china center for type culture Collection: the preservation number of the Chinese Wuhan university is: cctccc NO: m2013691, the preservation time is: 12 months 23 days 2013;

(3) Lactobacillus gasseri LG-G12 with the preservation unit: china center for type culture Collection: the preservation number of the Chinese Wuhan university is: cctccc NO: m2019829, the preservation time is: 10 months 17 days 2019;

(4) The bifidobacterium animalis subspecies BL-G101 are stored in the following units: china center for type culture Collection: the preservation number of the Chinese Wuhan university is: cctccc NO: m2013336, the preservation time is: 2013, 7, 16;

(5) Bifidobacterium longum BL-G301, deposited in the following units: china center for type culture Collection: the preservation number of the Chinese Wuhan university is: cctccc NO: m2013689, the preservation time is: 12 months 23 days 2013;

(6) Bifidobacterium infantis BI-G201, deposit unit: china center for type culture Collection: the preservation number of the Chinese Wuhan university is: cctccc NO: m2013688, the preservation time is: 12 months 23 days 2013;

all documents mentioned in this application are incorporated by reference as if each were individually incorporated by reference. Further, it will be appreciated that various changes and modifications may be made by those skilled in the art after reading the above teachings, and such equivalents are intended to fall within the scope of the claims appended hereto.

Claims

1. A composite formulation, the composite formulation comprising:

2. The composite formulation of claim 1, wherein the weight ratio of the probiotic formulation to the prebiotic formulation is 1:5-20, preferably 1:5-10.

3. The composite formulation of claim 1, wherein the prebiotic formulation comprises the following components in parts by weight:

4. The complex formulation of claim 1, wherein the prebiotic formulation further comprises inulin and/or galacto-oligosaccharides, and optionally isomaltose, soy oligosaccharides and lactulose.

5. The composite formulation of claim 4, wherein the prebiotic formulation comprises the following ingredients in parts by weight:

10-40 parts by weight, preferably 15-30 parts by weight, more preferably 20-25 parts by weight of galacto-oligosaccharide;

5 to 40 parts by weight, preferably 10 to 30 parts by weight, more preferably 15 to 25 parts by weight of inulin;

6. The composite preparation according to claim 1, wherein the viable count of lactic acid bacteria in the probiotic preparation is (0.1-20) ×10 ¹⁰ CFU/g, preferably (1-10). Times.10 ¹⁰ CFU/g, more preferably > 2.0X10 ¹⁰ CFU/g。

7. The composite formulation of claim 1, wherein the composite formulation comprises a probiotic formulation and a prebiotic formulation, packaged separately.

8. The composite formulation of claim 1, wherein the composite formulation is in the form of a powder, capsule, solution, suspension, emulsion, syrup, liquid spray, ointment, or gel.

9. Use of a complex formulation according to claim 1 for the preparation of a composition for the prevention and/or treatment of allergic rhinitis.

10. A product combination comprising a first package having the prebiotic formulation of claim 1 enclosed therein and a second package having the probiotic formulation of claim 1 enclosed therein.