CN106166295B

CN106166295B - Interferon oral preparation for treating viral diarrhea

Info

Publication number: CN106166295B
Application number: CN201610329710.8A
Authority: CN
Inventors: 陈全民; 潘海
Original assignee: Sciwind Biosciences Co Ltd
Current assignee: Hangzhou Xianweida Biotechnology Co ltd
Priority date: 2015-05-19
Filing date: 2016-05-18
Publication date: 2020-04-07
Anticipated expiration: 2036-05-18
Also published as: CN106166295A

Abstract

The invention provides an interferon oral enteric preparation, which consists of a blank layer, an interferon drug-containing layer and an enteric coating layer, and does not contain a protease inhibitor and/or an absorption enhancer.

Description

Interferon oral preparation for treating viral diarrhea

Technical Field

The invention relates to an interferon oral enteric preparation, in particular to an interferon oral enteric preparation with good prevention and treatment effects on viral diarrhea, and belongs to the technical field of biological pharmacy.

Background

Viral diarrhea

The incidence of infectious diarrhea in children is second only to respiratory infections, a global serious public health problem, especially in children under 5 years of age in developing countries, with the improvement of sanitary conditions and the use of antibiotics, the proportion of bacterial factors in children's infectious diarrhea is gradually reduced, and the proportion of diarrhea caused by viruses is increased. Rotavirus (RV) infection is a major cause of infantile diarrhea, norovirus (NoV) also occurs at a high rate, and other viruses such as sapovirus (SaV), enterovirus (EAdv), astrovirus (astrv)) also have a certain rate.

Rotavirus is the most common cause of severe dehydration diarrhea, and each year, more than fifty thousand children under 5 years of age die from rotavirus diarrhea. More than 80% of these rotavirus-related deaths occur in sub-saharan africa and in developing countries of south asia. In addition, rotavirus causes 25% -50% of all diarrhea hospitalizations in developing and developed countries, and outpatient healthcare receives over two thousand infants each year. Almost every child is infected with rotavirus before the age of 5 years.

Rotavirus diarrhea not only causes the burden of diseases in society, but also brings huge economic loss. Even in developed countries, rotavirus infections in infants are very common. U.S. studies have found medical costs associated with rotavirus infections as high as $ 3.52 to $ 10 billion per year. Immunization programs are practiced that save $ 0.79 million for health care and $ 4.66 million for other aspects of society. Chinese continental survey shows that: in regions with different economic levels, the clinic treatment cost of rotavirus diarrhea patients under the age of 5 years is 64-248 yuan each time, the hospitalization cost is 658-1920 yuan each time, the average clinic treatment cost of national rotavirus diarrhea is preliminarily estimated to be 100 yuan, and the average hospitalization cost is about 839 yuan.

To date, rotavirus infection has not been specifically treated, and treatment has mainly relied on fluid replacement for several days or probiotic preparations for regulating intestinal flora. More seriously, rotavirus infection is not related to sanitary conditions, and even if the sanitary conditions are improved, the spread of the virus cannot be effectively controlled.

Vaccine prophylaxis is considered the best preventive measure and vaccine immunization is the only viable approach to prevent the higher morbidity and mortality of rotavirus diarrhoea. Two live, orally available rotavirus vaccines are currently commercially available

(Merck and Co.Inc., PA, USA and Sanofi Pasteur MSD SNC, Lyon, France) and

(GSK Biologicals, Rixensearch, Belgium). In 2009, months 4 and 10, the WHO consulted additional efficacy data from different populations in africa and asia against a strategic consultant panel, as well as studies after U.S. approval, and promoted vaccination recommendations to all regions of the world.

Interferon

Interferons are an important family of cytokines, and have broad-spectrum antiviral, anti-cell-proliferation and immunomodulatory effects. Mammalian interferons can be classified into types I, II and III, and have been used to treat a number of diseases including viral infections (e.g., hepatitis C, hepatitis B, HIV), inflammatory disorders, and cancers (e.g., myeloma, lymphoma, liver cancer, breast cancer, melanoma, leukemia, etc.).

Interferons are of particular interest from a therapeutic standpoint (for a review of interferons see De Maeyer and DeMaeyer-Guignard, "interferons"; The Cytokine Handbook, third edition, Thompson (eds.), pages 491-516 (Academic Press Ltd.1998) and Walsh, Biopharmaceuticals: Biochemistry and Biotechnology, pages 158-188 (John Wiley & Sons 1998)). interferons exhibit a variety of biological activities useful in The treatment of certain autoimmune diseases and in enhancing immune responses to infectious agents including viruses, bacteria, fungi and protozoa.

The α chain of the human interferon α/β receptor comprises an N-terminal extracellular domain that has the characteristics of a class II cytokine receptor.

Type III interferons (also known as IL-28/IL-29) are a recently discovered family of novel proteins that share sequence homology with type I interferons and with IL-10 gene sequences. This new family is described in detail in commonly owned PCT application WO 02/086087 and Sheppard et al, Nature Immunol.4:63-68,2003, which are incorporated herein by reference in their entirety. Functionally, both IL-28 and IL-29, like type I interferons, induce an antiviral state in cells. However, the main difference between the interferon systems of type I and III is that they are directed against different sets and types of responding cells. Receptors for type I interferons, although not necessarily present on all nucleated cell surfaces, are found on most nucleated cell surfaces, whereas receptors for functional type III interferons are more preferentially expressed on epidermal cell surfaces (SommereynsC, Paul S, et al.

Interferon has been clinically used for treating viral diarrhea, but its administration is mostly by intramuscular injection, large dose aerosol inhalation, enema (expression, xu dahong, etc. clinical medical engineering 2014; dunjinli, clinical rational medication 2014; li, south china journal of medicine 2014; mare, journal of practical medicine 2006). The interferon injection is also clinically orally taken for treating the viral diarrhea, but the interferon injection is only diluted by corresponding water and then is orally taken.

Existing polypeptide/interferon oral formulations

One limiting factor in the use of proteins and polypeptides as pharmaceuticals is their metabolism by plasma proteins when administered parenterally. The oral route of administration presents further problems due to proteolysis in the stomach, acidic conditions therein or destruction of the molecule before reaching its intended target. For example, peptides and protein fragments produced by the action of gastric and pancreatic enzymes are cleaved by exopeptidases and endonucleases of the intestinal villus marginal membrane to produce dipeptides and tripeptides. If proteolysis by pancreatin can be avoided, the polypeptides are degraded by villous edge peptidases. Polypeptides or proteins that may also remain through the stomach are metabolized in the intestinal mucosa, where penetration barriers prevent their entry into the cells.

In order to remain in the stomach and intestinal tract until absorption through the intestinal mucosa, the molecule is typically formulated in a protective dosage form. For example, the protein may be administered with a protease inhibitor (e.g., bacitracin or a derivative thereof, soybean trypsin or aprotinin) and stabilized with a polymeric material. While the intestinal mucosa barrier is passed by adding absorption enhancer (such as sodium salicylate, sodium lauryl sulfate, oleic acid, linoleic acid, lecithin, etc.). However, the enzyme inhibitor can effectively avoid the hydrolysis of protease, but the enzyme inhibitor also influences the digestion of food protein, and the long-term administration of the enzyme inhibitor can cause the disturbance of digestion and absorption and even the enlargement or hyperplasia of pancreas; the absorption enhancer can cause irreversible damage to cell membranes while increasing intestinal permeability, and can cause membrane poisoning after long-term administration.

Another treatment is to avoid the gastrointestinal tract altogether, by delivering proteins to the oropharyngeal portion in the form of lozenges or by holding the solution for a period of time in the mouth, through the mucosa and into the blood circulation. US5824300 discloses the use of an oral interferon for the treatment of neoplastic disease, in the form of a lozenge which can be buccal by absorption through the mucous membranes of the patient's pharynx. WO2013/165963 discloses a method of treating thrombocytopenia with interferon orally, by administering an amount of interferon to a thrombocytopenic patient sublingually in a saliva-soluble lozenge. US20070237723 discloses a method for treating chronic cough by oral administration of interferon, wherein interferon is inhaled by nebulization from oral cavity by patient to achieve antiviral effect and treat cough.

Protein drugs such as interferon are prepared into oral preparations, and the clinical treatment effect is fully ensured, protease inhibitors and/or absorption promoters seem to be indispensable auxiliary agents, but the auxiliary agents bring corresponding safety hazards (causing digestive absorption disorder or membrane poisoning due to cell membrane damage), and the development of the oral administration route of the protein drugs such as interferon is greatly limited by the existence of the factors.

Disclosure of Invention

The present inventors have surprisingly found, in the study of viral diarrhoeal diseases, that oral administration of a specific interferon formulation provides a very good preventive and/or therapeutic effect against diarrhoea caused by viruses.

Therefore, an object of the present invention is to provide an interferon preparation for oral administration for preventing or treating viral diarrhea, which can be administered orally to a patient infected with enterovirus without injection, and can achieve a good preventive or therapeutic effect, which is equivalent to or even better than that of injection, and which greatly facilitates administration to the patient.

An interferon formulation for use in the prevention or treatment of the present viral diarrhea, preferably an oral interferon formulation in an enteric form, such as enteric coated particles of interferon.

The interferon enteric-coated particles of the present invention are a type that does not contain a protease inhibitor, for example, to prevent the hydrolysis thereof by an enzyme, and does not contain an absorption enhancer, for example, additionally added to solve the intestinal mucosal barrier. Although the oral interferon enteric preparation (such as dry suspension) of the present invention does not contain protease inhibitor and absorption enhancer, it has a therapeutic effect equivalent to that of injection administration in preventing or treating viral diarrhea diseases, and such a therapeutic effect is greatly beyond the expectation of the skilled person.

The interferon enteric-coated particles are stable in long-term storage under certain conditions, and are convenient for patients and improve the compliance of the patients due to the fact that the interferon enteric-coated particles are directly orally taken.

Specifically, the interferon enteric-coated particles comprise: the enteric-coated pellet comprises a blank pellet core, an interferon layer coated on the blank pellet core, and an enteric-coated layer coated on the interferon layer.

The blank pellet core is a blank pellet core (the diameter is in a range of 10 nanometers to 500 micrometers) with a small particle size, for example, the blank pellet core can be a nano pellet core, and the material of the pellet core can be sucrose, starch, microcrystalline cellulose, biodegradable polymer and the like.

The interferon layer coated on the blank pellet core is obtained by dissolving an interferon stock solution in a proper solvent and uniformly coating the interferon stock solution on the blank pellet core by means of spraying or evaporation and the like.

The enteric coating coated on the interferon layer is preferably soluble at pH5.0-6.8, such as Cellulose Acetate Phthalate (CAP), polyvinyl acetate titanate (PVAP), and acrylic resin (such as methyl methacrylate), and the coating weight can be 0.5-10% of the weight of the pill core.

The interferon microparticles obtained above can be mixed with other suitable excipients such as diluent, filler, lubricant, glidant, etc., and compressed into tablet; or directly filling the obtained interferon enteric-coated particles into capsules to obtain capsules; or mixing the obtained interferon enteric-coated particles with a certain sweetener and the like, packaging in a bag, preparing a dry suspension, and suspending in water before oral administration; or suspending the obtained interferon enteric-coated particles in a proper solution to prepare oral liquid. Preferably, the interferon enteric-coated particles are a dry suspension.

Animal experiment research shows that the interferon enteric-coated particles provided by the invention are orally taken, and through the index investigation of diarrhea rate/severe diarrhea rate, diarrhea duration days and HRV antigen positive duration days, the therapeutic effect of the interferon enteric-coated particles on viral diarrhea is equivalent to or even better than that of interferon with the same dose through intramuscular or subcutaneous injection.

In the invention, the interferon can be type I interferon, such as interferon α, interferon β, interferon omega, interferon delta and interferon tau, type II interferon, such as interferon gamma, type III interferon, such as interferon lambda 1, interferon lambda 2 and interferon lambda 3, preferably, the interferon is type III interferon.

In the present invention, the viral diarrhea may be diarrhea caused by rotavirus, norovirus, zavirus, enteroadenovirus, and astrovirus, for example.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

Example 1 preparation of Interferon enteric particles-method one

200 g of sucrose pellet core with the diameter of 200-400 microns is taken and placed in a fluidized bed, 50 ml of type III interferon (IFN-lambda 1) solution (containing 100 mg of interferon) is subjected to spray drying in a side spraying mode, the inlet temperature is 40-45 ℃, and the drying time is 30 minutes. After the spray drying is finished, the interferon is coated on the pill core to form interferon particles. The interferon particles were spray dried in a side-spray manner using a solution of Ewing's L30D-55 in ethanol, also in a fluidized bed, at an inlet temperature of 35-40 deg.C for a drying time of 25 minutes. And (3) obtaining the sandwich-type interferon enteric-coated particles after drying, namely, the innermost layer is a sucrose pill core, the middle layer is the three-type interferon, and the outermost layer is an enteric coating layer. The particle size of the sandwich interferon particles is 250-450 microns (about 50 microns larger than that of the sucrose pellet core). The unit mass of the interferon microparticles contains 0.3-0.4 mg/g of the type III interferon. The microparticles are cultured at 37 ℃ for 2 hours under an acidity condition close to that of the stomach (pH2.0) without releasing the type III interferon, and cultured at 37 ℃ for 8 hours under a pH condition of pH5.5 or higher, over 85% of the type III interferon contained in the microparticles is released.

EXAMPLE 2 preparation of Interferon enteric particles-method two

200mg of pharmaceutical-grade silica fine particles having an average particle diameter of about 20nm were suspended in 20ml of purified water, and 5ml of a type III interferon solution (IFN-. lambda.1, 4mg/ml) was mixed therewith and vacuum-dried with stirring. When all the water was removed, silica fine particles having type III interferon adhered to the surface were obtained. The resulting microparticles were suspended in 20ml of ethanol, and 20mg of enteric material, eucalyptus L100, was dissolved in the above ethanol, and the mixture was placed under vacuum while being subjected to ultrasonic treatment until all the liquid was removed to obtain a dry powder. And (3) crushing the obtained dry powder, and sieving to remove larger particles to obtain target enteric soluble tri-type interferon particles, wherein the average particle size of the particles is 0.5-5 microns. The unit mass of the particles obtained by the method contains 50mg/g of the type III interferon. The microparticles were incubated at 30 ℃ for 2 hours in buffer pH2.0 to release 20-30% of the type III interferon, and at 37 ℃ for 8 hours at pH7.0 to release > 80% of the type III interferon.

By the same process steps as in examples 1 and 2, enteric microparticles of IFN-. lambda.2 or IFN-. lambda.3 can be prepared

EXAMPLE 3 therapeutic Effect of enteric-coated formulations on Robotic mice infected with rotavirus

Rotavirus: HRV virus strains were provided by the institute for viruses of the chinese academy of preventive medicine sciences. The cells used for virus propagation and assay were MA104 cells (African green monkey fetal kidney cell passage cell line).

Medicine preparation: IFN-lambda 1 is prepared by itself and prepared into injection and enteric-coated particles respectively (example 1 method); IFN-. lambda.2 was self-prepared and made into enteric particles as in example 2.

Experimental animals: bal/c suckling mice, SPF grade, provided by Beijing, are delivered naturally, 88 suckling mice are born in 8 litters, the weight is 2.3-2.8 g, the suckling mice are raised in a suckling mouse cage with an air filter membrane, the postnatal mother mice are fed with milk, the raising environment is 12 hours of light, 12 is dark, the temperature is 18-25 ℃, the humidity is 40% -70%, and the experimental environment is SFP grade.

Group A rotavirus diagnostic kit: purchased from Beijing Wan peptide biopharmaceutical Co., Ltd.

Experimental methods

(1) Grouping

84 suckling mice were selected for the experiment, and were divided into 7 groups with the mother mice, each group consisting of 12 mice: HRV infection group (group A), IFN-lambda 1 injection pretreatment group (group B), IFN-lambda 1 enteric particle pretreatment group (group C), IFN-lambda 1 injection treatment group (group D), IFN-lambda 1 enteric particle treatment group (group E), IFN-lambda 1 solution treatment group (group F), and IFN-lambda 2 enteric particle treatment group (group G). HRV infection group reference (Xushiwei, et al. Chinese journal of clinical infection 2010,3(2):100) method to establish HRV infection model of suckling mice, i.e. 50 μ l HRV solution (3 × 10) orally administered (naturally sucked) by modified 1ml syringe at birth 4d of suckling mice⁶pfu/ml). IFN-lambda 1 injection pretreatment group performs intraperitoneal injection of 50 mu l (3 ten thousand IU/one) of IFN-lambda 1 every day from day 1 after birth, and IFN-lambda 1 enteric-coated microparticle pretreatment group performs oral administration of 50 mu l of enteric-coated microparticle suspension (3 ten thousand IU/one) from day 1 after birth, continuously performs 3d, and performs oral administration of 50 mu l of HRV solution (3 multiplied by 10) from day 4⁶pfu/ml), 50. mu.l HRV solution (3X 10) was orally administered at 4d of each treatment group⁶pfu/ml), 1h later, 50 mu of IFN-lambda 1 liquid is injected into the abdominal cavityl (3 ten thousand IU/piece), 50 mul (3 ten thousand IU/piece) of IFN-lambda 1 enteric-coated particle suspension liquid is orally administrated, 50 mul (3 ten thousand IU/piece) of IFN-lambda 1 injection liquid is orally administrated, 50 mul (3 ten thousand IU/piece) of IFN-lambda 2 enteric-coated suspension liquid is orally administrated, and the continuous time is 4 days.

2. And (3) clinical observation: successive clinical observations were made on suckling mice starting on day 4, 2 times a day (1 in the morning and evening): weighing the weight of the suckling mouse, observing the diarrhea condition of the suckling mouse, counting the death number of the suckling mouse, and detecting the HRV antigen in the response of the suckling mouse by an ELISA method according to the instruction of the reagent. The literature on judgment of diarrhea (Takahashi K, Ohashi K, et al, Antichronologically Agents Chemother.2002,46:420) is classified in 6 grades: level 1 is no stool, level 2 is yellow formed stool, level 3 is yellow pasty stool, level 4 is yellow water-like mucus stool, level 5 is yellow egg soup stool, and level 6 is completely yellow water-like stool. The diarrhea was judged on grade 3 or above, light on grade 3, 4, 5, and heavy on grade 6.

3. Detection of RV antigens in infected animal responses: and (3) putting a proper amount of the complex sample into a container, adding 9 times of normal saline, stirring and uniformly mixing for 2 minutes, dropwise adding 3-4 drops of excrement supernatant at the sample adding position of the kit after centrifugation, horizontally placing at room temperature, and judging the result within 10 min.

Table 1: weight change (g) of suckling mice in each group

Group of	n	Postnatal day 4	Postnatal day 9
				A	12	3.72±0.26	6.28±0.71
B	12	3.55±0.31	6.76±0.64^*
				C	12	3.68±0.29	6.98±0.73^**
D	12	3.82±0.37	6.46±0.49
				E	12	3.76±0.27	6.67±0.62^*
F	12	3.87±0.34	6.41±0.65
				G	12	3.79±0.29	6.59±0.54^*

Compared with the group A, the method has the advantages that,^*P<0.05，^**P<0.01.

table 2: diarrhea in all groups of suckling mice

Group of	n	Severe diarrhea	Mild diarrhea	Death was caused by death	The rate of severe diarrhea%	The diarrhea rate%
							A	12	9	3	3	75	100
B	12	0	3	0	0	25
							C	12	0	2	0	0	16.7
D	12	1	5	0	8.3	50
							E	12	0	5	0	0	41.7
F	12	5	6	0	41.7	91.7
							G	12	0	6	0	0	50

Table 3: average days of diarrhea and HRV antigen Positive days (day d) for all groups of suckling mice

Group of	n	Mean days of diarrhea d	Mean number of HRV antigen positivity days d
				A	12	4.72±0.66	5.48±0.68
B	12	2.05±0.51^**	3.16±0.56^**
				C	12	1.89±0.49^**	2.57±0.43^**
D	12	2.92±0.52^*	3.86±0.62^**
				E	12	2.76±0.67^*	3.57±0.56^**
F	12	3.99±0.54	4.71±0.70
				G	12	2.49±0.59^*	3.36±0.54^**

P <0.05, P <0.01 compared to group a

Body weight change in suckling mice: by day 6 after viral infection (postnatal day 9), the weight gain of each treatment group was superior to that of the untreated HRV suckling mouse group, wherein the weight gains of the IFN-lambda 1 pretreatment group, the IFN-lambda 1 and the IFN-lambda 2 enteric particle treatment group were more obvious than that of the HRV untreated group, and the differences were significant. The differences of the weight gains of the IFN-lambda 1 injection treatment group and the IFN-lambda 1 injection oral treatment group are not significant.

Diarrhea status in suckling mice: the HRV-infected suckling mice have symptoms of mental retardation and reduced activity in the next day after the feeding of the virus, diarrhea symptoms appear in 3-4 days after the virus infection, the diarrhea rate reaches 100%, 9 severe diarrhea and 3 mild diarrhea are yellow egg-shaped feces, and 3 suckling mice die in the 5 th day after the virus infection. All suckling mice in the group pretreated by the IFN-lambda 1 injection and the IFN-lambda 1 enteric-coated particles do not have severe diarrhea symptoms, 3 suckling mice in the injection group have mild diarrhea symptoms after virus infection in 3-4 days, and 2 suckling mice in the enteric-coated particles have mild diarrhea symptoms which are yellow watery mucus stools, and the rest suckling mice have good mental status, normal activities and no wrinkles on the skin. In the whole process of FN-lambda 1 injection, IFN-lambda 1 and IFN-lambda 2 enteric-coated particle treatment groups, nearly galactophore mice have diarrhea symptoms, wherein 1 of the mice in the injection treatment group have severe diarrhea symptoms, 5 mice have mild diarrhea, and the mice with mild diarrhea mainly have yellow watery mucus stool to yellow egg-shaped stool. Diarrhea symptoms also appear in the whole process of the IFN-lambda 1 injection oral administration treatment group, wherein 5 suckling mice have severe diarrhea symptoms, and the rest 6 suckling mice have mild diarrhea symptoms. None of the suckling mice died in the treatment group and the prevention group. The oral administration treatment of the IFN-lambda enteric particles is slightly better than the IFN-lambda injection administration group regardless of the preventive administration or the therapeutic administration, and although the difference between the oral administration treatment group and the IFN-lambda injection administration group is not significant, the oral administration treatment group and the IFN-lambda injection administration treatment group are obviously better than the IFN-lambda injection administration treatment group, and the difference between the oral administration treatment group and the IFN-lambda injection administration treatment group is significant.

Duration of diarrhea and HRV antigen positive days in suckling mice: starting from day 3 of virus infection, the positive days of HRV antigen in the stool of each group of diarrhea suckling mice are counted. Compared with the HRV infected group, the diarrhea duration days and the HRV antigen positive days of the suckling mice in each treatment group are obviously shortened, and the difference is significant (P <0.05 and P <0.01), but the IFN-lambda 1 injection oral treatment group is not capable of obviously reducing the diarrhea duration days and the HRV antigen positive days of the suckling mice except the IFN-lambda 1 injection oral treatment group. In each treatment group, the IFN-lambda 1 injection and the enteric particle pretreatment group are shortened more obviously, and next, the IFN-lambda 1, the IFN-lambda 2 enteric particles and the IFN-lambda 1 injection treatment group are provided, the oral treatment effect of the IFN-lambda enteric particles is equivalent to or even better than that of the IFN-lambda 1 injection treatment group, but the difference is not obvious.

The results show that for the HRV infected suckling mice, IFN-lambda can achieve the effect equivalent to or even better than the injection mode in terms of body weight, diarrhea rate/severe diarrhea rate, diarrhea days and HRV positive days, and the treatment effect of the IFN-lambda injection directly orally administered on the suckling mice with enterovirus diarrhea is obviously reduced.

Example 4 stability study

The storage condition of the interferon injection is 2-8 ℃, and various influence factors are tested on the preparation in consideration of the particularity of the IFN-lambda 1 enteric particle dosage form, so that the stability of the interferon injection under the conditions of strong light, high temperature and high humidity is inspected, and an experimental basis is provided for the storage condition of the interferon injection. Accelerated and long-term stability tests were performed simultaneously. The stability test refers to Chinese pharmacopoeia, and appearance, titer, pH and release degree are selected as evaluation standards.

The IFN-lambda 1 enteric-coated particles obtained in example 1 were tested for stability under the conditions of 4500Lx +/-500 Lx (refrigerated dry), high humidity RH 75% (refrigerated dark) and high temperature 40 ℃ (dark dry), and the obtained results show that: the product is very sensitive to temperature, and the potency of the product is reduced to 70% of that of 0 day at 15 days at high temperature (40 ℃), so that the interferon enteric-coated particles are stored by refrigeration (2-8 ℃). Under high humidity environment (75%), the particles are easy to absorb moisture and block, so it is recommended to use the composite bag material for medicine to package, which can isolate the moisture in the air.

The result of the accelerated stability test shows that the product is packaged by adopting a moistureproof medicinal composite bag, the moisture-proof medicinal composite bag is placed for 6 months under the conditions that the temperature is 25 +/-2 ℃ and the relative humidity is 60 +/-10 percent, the titer has a descending trend but still is within a qualified range, and other tests have no significant difference compared with the test in 0 month, so that the selected inner packaging material has no influence on the quality of the product, the sealing property and the light shielding property of the product meet the requirements, and the inherent quality of the product is not influenced. Meanwhile, the product is sensitive to temperature and should be refrigerated for storage. Furthermore, the moisture-proof medicinal composite bag is adopted for packaging, and the indexes of the potency, the pH value and the dissolution rate are almost not changed after long-term test for 12 months under the conditions that the temperature is 6 +/-2 ℃ and the relative humidity is 60 +/-10%, which shows that the product is stable under the conditions.

By combining the test results of the above-mentioned influence test, acceleration and long-term stability, the condition of recombinant human interferon lambda enteric-coated particles is that the recombinant human interferon lambda enteric-coated particles are preserved at 2-8 ℃ in the dark, and the effective period can be tentatively set to 18 months.

Dissolution rate

Culturing at 37 deg.C for 2 hr under acidity condition (pH2.0) close to that of stomach, wherein the dissolution rate of IFN- λ 1 is not more than 15%, and culturing at 37 deg.C for 8 hr under pH7.0 condition, wherein more than 85% of IFN- λ 1 contained in the microparticles is released.

Claims

1. An interferon enteric-coated particle for treating viral diarrhea, the enteric-coated preparation comprises a blank layer, an interferon drug-containing layer coated on the blank layer, and an enteric-coated layer coated on the interferon drug-containing layer, and the interferon enteric-coated particle does not contain an absorption enhancer, the interferon is type III interferon, the type III interferon is selected from interferon lambda 1, lambda 2 or lambda 3, the enteric-coated layer is an enteric-coated layer which is soluble at a pH higher than 5.5, and the enteric-coated layer is selected from cellulose acetate phthalate, polyvinyl acetate titanate or acrylic resin.

2. The enteric interferon microparticle of claim 1, wherein the blank layer is a blank pellet core with a particle size of 10nm to 500 μm, and the material of the pellet core is selected from sucrose, starch, or biodegradable polymer.

3. The enteric interferon microparticle of claim 2, wherein the biodegradable polymer is microcrystalline cellulose.

4. The enteric interferon microparticle of claim 1, wherein the weight gain of the enteric coating is 0.5-10% of the weight of the pellet-containing core.

5. The enteric interferon microparticle of claim 1, wherein the viral diarrhea is diarrhea caused by rotavirus or norovirus.

6. Use of enteric coated microgranules comprising an interferon according to any of claims 1 to 5 for the manufacture of an oral medicament for the prevention or treatment of viral diarrhea.

7. The use according to claim 6, wherein the oral medicament comprises enteric particles of interferon according to any of claims 1 to 4, together with a pharmaceutically acceptable excipient.

8. The use of claim 7, wherein the oral medicament is an oral granule, tablet, capsule, or suspension.