CN113893333B - Insulin slow-release oral patch and preparation method and application thereof - Google Patents

Insulin slow-release oral patch and preparation method and application thereof Download PDF

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CN113893333B
CN113893333B CN202111210673.6A CN202111210673A CN113893333B CN 113893333 B CN113893333 B CN 113893333B CN 202111210673 A CN202111210673 A CN 202111210673A CN 113893333 B CN113893333 B CN 113893333B
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insulin
solution
preparation
water phase
nano particles
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CN113893333A (en
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薛建鹏
徐寒梅
王瑱
胡加亮
杜南星
黄锦辉
李则青
赵文静
来志龙
赖文婷
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China Pharmaceutical University
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China Pharmaceutical University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2013—Organic compounds, e.g. phospholipids, fats
    • A61K9/2018—Sugars, or sugar alcohols, e.g. lactose, mannitol; Derivatives thereof, e.g. polysorbates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/2027—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/2031—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyethylene oxide, poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2054—Cellulose; Cellulose derivatives, e.g. hydroxypropyl methylcellulose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2095—Tabletting processes; Dosage units made by direct compression of powders or specially processed granules, by eliminating solvents, by melt-extrusion, by injection molding, by 3D printing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics

Abstract

The invention relates to the field of biological medicine, in particular to an insulin slow-release oral patch, and a preparation method and application thereof. A pharmaceutical formulation characterized in that: the medical auxiliary material comprises nano particles and medical auxiliary materials, wherein the medical auxiliary materials comprise a filler, an adhesive and a lubricant; the dosage form of the pharmaceutical preparation is an oral patch. The invention prepares the insulin into the sustained-release oral patch, which not only avoids the first pass effect of the gastrointestinal tract, prolongs the action time of the medicine, improves the curative effect and economic benefit of the medicine, but also reduces the influence of enzymes on the insulin, and avoids the pain caused by injection of patients, and the prepared tablet has strong adhesive capability, good release and moderate dissolution rate.

Description

Insulin slow-release oral patch and preparation method and application thereof
Technical Field
The invention relates to the field of biological medicine, in particular to an insulin slow-release oral patch, and a preparation method and application thereof.
Background
Insulin is a protein hormone secreted by islet beta cells within the pancreas by stimulation with endogenous or exogenous substances such as glucose, lactose, ribose, arginine, glucagon, and the like. Insulin is the only hormone in the body that reduces blood glucose while promoting glycogen, fat, protein synthesis. Exogenous insulin is mainly used to treat diabetes. Is a main medicine for treating type 1 diabetes. Good effect, but harsh preservation conditions, easy inactivation and instability.
For the above reasons, the clinical application requires frequent administration of injections, which can cause pain and complications to the patient. It is a complex and difficult task to prepare it into a stable, safe and efficient pharmaceutical formulation. Numerous studies are currently being conducted on strategies for delivery of polypeptide protein drugs, and a variety of novel drug delivery systems for different routes of administration have been reported. The nano medicine carrying system is a medicine carrying mode with potential advantages due to the characteristics of the nano medicine carrying system.
The nanoparticle has the particle size of 1-1000nm, good stability and good permeability, can prolong the half life of the drug, can be used as a substitute for unstable liposome and vesicle, can be used as an initial material for preparing nanocapsules and nanospheres, and can protect the activity of the drug and realize the slow release effect of the drug. These advantages make the nanoparticle widely used in the fields of beauty industry, agriculture, chemical industry, pharmacy, etc.
The insulin nanoparticles can prolong the in vivo half-life of insulin, reduce clearance rate and immunogenicity and antigenicity, reduce administration frequency, and thus can improve treatment compliance.
The oral administration is the most acceptable administration method because of the simple and convenient painless characteristics, so we further prepare the insulin nanoparticle lyophilized powder into oral patch tablets for administration through oral mucosa. The insulin nano particles can be released from the tablets into blood for slow release due to good membrane permeation performance, so that the half life of the medicine in vivo is prolonged, the action time of the medicine in vivo is prolonged, meanwhile, the pain and the harm left by the body when the patient is injected are eliminated by oral administration, the treatment compliance of the patient is improved, the use of the medicine can be reduced, and the cost performance is high.
Disclosure of Invention
The invention aims to provide the nano-particles containing the insulin with a double-layer structure, which not only have a good slow release effect, thereby realizing the function of releasing the medicine for a long time, but also are beneficial to the transmembrane movement of the nano-particles due to the positive charges carried on the outer layer of the nano-particles, thereby increasing the intake and the transportation of the medicine. The nanoparticle can be mixed with medicinal adjuvants to prepare into pharmaceutical preparation.
The second purpose of the invention is to provide an oral patch tablet prepared from the insulin nanoparticle freeze-dried powder and a preparation method thereof.
The present disclosure provides a nanoparticle having a bilayer structure, comprising:
(a) An outer layer having a positive charge; and
(b) A biodegradable polymer inner layer surrounded by an outer layer;
the solid insulin is uniformly distributed in the inner polymer layer;
the outer layer of the insulin nanoparticle comprises chitosan, polydopamine, polylysine, polyarginine and polyhistidine, so that the charge of the nanoparticle is increased, the nanoparticle is positively charged, the combination with negatively charged cell membranes is facilitated, and further the transmembrane movement of the nanoparticle is facilitated, so that the intake and the transportation of insulin medicine are increased.
In an embodiment, the biodegradable polymer inner layer composition comprises a polylactic acid-glycolic acid copolymer, polyvinyl alcohol, and sodium alginate.
In embodiments, the weight of the solid insulin is 0.1 to 0.5wt% of the total weight of the nanoparticle;
in a preferred embodiment, the weight of the solid insulin is 0.15-0.4wt% of the total weight of the nanoparticle. More preferably, the weight of the solid insulin is 0.2-0.3wt% of the total weight of the nanoparticle.
In a preferred embodiment, the nanoparticle has a particle size of 10-1000nm; and/or the nanoparticle has a PDI of less than 0.3;
in a preferred embodiment, the nanoparticle outer layer has a thickness of 12-22nm.
The invention provides a preparation method of the nano-particles, which comprises the following steps:
1) Dissolving solid insulin powder in acetate buffer to form insulin solution
2) Dispersing insulin solution in sodium alginate solution to serve as an inner water phase, mixing the inner water phase with an organic phase, and performing ultrasonic emulsification to form water-in-oil colostrum;
3) Injecting the colostrum into an external water phase, fully and uniformly mixing, and performing ultrasonic emulsification to form a water-in-oil-in-water compound emulsion solution, wherein the external water phase also contains polyvinyl alcohol and Ca 2+ A chitosan;
4) And volatilizing the organic solvent of the re-emulsion solution completely, and forming nano particles after freeze drying.
Wherein, the sodium alginate in the step 2) is helpful for dissolution and dispersion, and can promote the dissolution and dispersion of the sample, and can be used as a surfactant to promote the formation of colostrum with the inner water phase and also can be used as a surfactant to promote the formation of Ca with the outer water phase 2+ The reaction forms gel, helps the medicine to be wrapped in the nano particles, improves the encapsulation efficiency, and is easy to disperse after freeze-drying.
In a preferred embodiment, the organic phase is prepared by: dissolving solid polylactic acid-glycolic acid copolymer in an organic solvent to form an organic phase; and/or the organic solvent of the multiple emulsion solution is completely volatilized by low-speed stirring or reduced pressure distillation at room temperature.
In a preferred embodiment, the specific preparation method of the nanoparticle comprises the following steps:
(1) Preparing polylactic acid-glycolic acid copolymer solution: dissolving solid polylactic acid-glycolic acid copolymer in an organic solvent to form an organic phase;
(2) Insulin dispersion solution preparation: dissolving solid insulin in acetate buffer solution to prepare insulin solution, and uniformly dispersing the insulin solution in sodium alginate solution to form an inner water phase;
(3) Preparation of colostrum: fully mixing the internal aqueous phase and the organic phase, and performing ultrasonic emulsification to form W/O colostrum;
(4) And (3) preparation of compound emulsion: injecting colostrum into a container containing polyvinyl alcohol and Ca at a certain concentration 2+ And the chitosan outer water phase are fully and uniformly mixed, and the ultrasonic emulsification is performed to form W/O/W compound emulsion;
(5) Nanoparticles: the organic solvent of the W/O/W compound emulsion is volatilized completely, and then freeze drying treatment is carried out.
In a preferred embodiment, the external aqueous phase consists of 0.0001% to 20.000% (w/v) PVA, 0.0001% to 3.000% (w/v) chitosan solution, 0.0001% to 0.18 (mol/v) Ca 2+ Mixing the solutions to form;
the organic solvent comprises one or more of dichloromethane, ethyl acetate and acetone; and/or the organic phase to internal aqueous phase volume ratio is (2-40): 1, the volume ratio of the colostrum to the external water phase is 1: (3-20);
in a preferred embodiment, the organic phase to inner aqueous phase volume ratio is 5:1 and the colostrum to outer aqueous phase volume ratio is 1:7.
In a preferred embodiment, the sodium alginate solution has a mass to volume ratio of 0.0001% to 50.000% (w/v).
In a preferred embodiment, the ultrasound conditions in step 3) are 10-1000w,1-60min ultrasound; the ultrasonic condition of the compound emulsion in the step 4) is 10-1000w and 1-60min ultrasonic.
In a preferred embodiment, in step 5), the multiple emulsion solution is stirred at low speed at 4 ℃ or vacuum spin evaporated at room temperature until the organic solvent is completely evaporated. Stirring at 1000-5000rpm for 4-12 hr or steaming at 25deg.C for 30-60 min; the freeze drying treatment steps are as follows: adding the nano particles into a mannitol solution containing 5% -7% (m/v), pre-freezing, and freeze-drying for 48 hours to obtain white powder with uniform texture.
The present disclosure provides a pharmaceutical formulation comprising the insulin nanoparticle. The insulin nano-particles can be prepared into different dosage forms according to requirements, and can play roles of prolonging the in-vivo half-life of insulin, reducing clearance rate, immunogenicity and antigenicity, reducing administration frequency and improving tolerance of patients. For example, it can be prepared into insulin injections, insulin nasal drops, insulin oral preparations or other pharmaceutically acceptable dosage forms as required.
In a preferred embodiment, the pharmaceutical formulation is in the form of an oral patch.
In a preferred embodiment, the pharmaceutical preparation comprises nanoparticles and pharmaceutical excipients, wherein the pharmaceutical excipients comprise a filler, an adhesive and a lubricant, and the mass ratio of the nanoparticles to the pharmaceutical excipients is (4-3): (1-2).
In a preferred embodiment, the mass ratio of the nanoparticle to the pharmaceutical excipients is 4:1, a step of; and/or, based on the total weight of the oral patch, the total weight ratio of the filler in the pharmaceutical auxiliary material to the oral patch is 2:25, the total weight ratio of the adhesive to the oral patch is 2:25, and the total weight ratio of the lubricant to the oral patch is 1:500; the oral patch is a tablet with the diameter of 5-10 mm, the thickness of 1-3 mm and the weight of 30-200 mg. The tablet has good adhesion, good release, and moderate dissolution rate.
In a preferred embodiment, the freeze-dried white powder is mixed with the oral patch auxiliary material in a ratio of 4:1, and the mixture is directly tabletted by a dry method to obtain a slow-release oral patch with good adhesion and good drug release, wherein the tablet weight is about 50mg, and the width is 6 mm and the thickness is 2 mm.
In preferred embodiments, the filler comprises starch or mannitol and/or the adhesive comprises one or more combinations of carbomers, carboxymethyl cellulose, sodium carboxymethyl cellulose; and/or the lubricant comprises magnesium stearate or talc; and/or the nanoparticles are selected from white freeze-dried powder formed by freeze-drying with a freeze-drying protective agent.
In a preferred embodiment, the present disclosure provides a method of preparing a pharmaceutical formulation formed from nanoparticles: is prepared by mixing nano particles and pharmaceutic adjuvant according to a certain proportion and tabletting, wherein the pharmaceutic adjuvant is a mixture containing a filler, an adhesive and a lubricant.
In a preferred embodiment, the present disclosure provides the use of the nanoparticle in the preparation of a pharmaceutical formulation.
In a preferred embodiment, the present disclosure provides the use of the nanoparticle in the preparation of an oral patch.
When the prepared oral patch acts, the insulin nano-particles can be released from the tablet into blood for slow release.
Principle of action
The main component of the polymer inner layer in the present disclosure is polylactic acid-glycolic acid copolymer (PLGA), also known as lactide-glycolide, polyglycolide and the like. It is polymerized by glycollic acid (or glycollic acid) and lactic acid, and is a biodegradable high-molecular material. The polylactic acid-glycolic acid copolymer material has a slow release function, and is nontoxic to human body after degradation. On the other hand, the nano particles disclosed by the invention have the advantages that the outer layer and the inner layer are biodegradable substances, and the nano particles have a slow release effect, so that the medicine is slowly released. Specifically, the outer layer of the material can be hydrolyzed in blood and can be degraded by in vivo enzymes so as to be metabolized, the inner layer of PLGA can be slowly degraded at the body temperature, and the inner layer of the PLGA is softer under the body temperature condition, so that the medicine is exchanged into the blood due to the action of osmotic pressure, and the medicine is slowly released by virtue of the degradation of the PLGA after the osmotic pressure is balanced, thereby achieving the characteristic of releasing the medicine for a long time. The nano particles can achieve the purpose of releasing the medicine for a long time by virtue of the coordination of the inner layer and the outer layer. The insulin nano-particles prepared by the invention can prolong the in vivo half-life of insulin, reduce the clearance rate, immunogenicity and antigenicity, reduce the administration frequency and improve the tolerance of patients.
Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
(1) Insulin, a special endogenous hormone, is very easily degraded in the gastrointestinal part, and patients with long-term insulin injections often suffer muscle pain, even partial muscle necrosis, and patient compliance is poor, so that the patch of the selected port is a good choice. However, how to ensure that insulin in the oral patch can be smoothly released to blood for generating action is still a difficult problem because insulin has poor fat solubility and is difficult to enter cortical cells, so that the insulin can stably enter blood circulation. Experiments prove that the oral liquid can be absorbed by an oral cavity inner membrane and enter blood, so that the blood sugar reducing effect is achieved, and the blood sugar reducing effect is stable.
(2) The insulin nanoparticle prepared by the invention has simple and stable process, high feasibility and industrialization, and the nanoparticle has round shape, a double-layer structure, smooth surface, good fluidity, PDI of less than 0.3, particle size of 10-1000nm, encapsulation rate of about 60 percent and in-vitro drug release performance meeting the characteristic of a long-acting preparation; on the other hand, the characteristic that the outer layer of the nano-particle has positive charges is favorable for being combined with negatively charged cell membranes, so that the transmembrane movement of the nano-particle is facilitated, and the intake and the transportation of insulin drugs are increased.
(3) According to the invention, the biodegradable material is used as a carrier, insulin is wrapped to prepare nano particles, and the nano particles are further prepared into the oral patch, so that the oral patch can increase the stability of the medicine, increase the half life of the medicine and prolong the acting time of the medicine due to small particle size and stable dispersion of the nano particles, thereby reducing the administration times and the dosage, improving the curative effect of the medicine, and being safer and more convenient by using an oral administration mode. On the other hand, the invention prepares the insulin into the sustained-release oral patch, which not only avoids the first pass effect of the gastrointestinal tract, prolongs the action time of the medicine, improves the curative effect and economic benefit of the medicine, but also reduces the influence of enzymes on the insulin, and avoids the pain caused by the injection of patients, and the prepared tablet has strong adhesive capacity, good release and moderate dissolution rate.
Drawings
FIG. 1 is a scanning electron microscope image of insulin nanoparticles of the present invention;
FIG. 2 is a graph showing the particle size distribution of insulin nanoparticles of the present invention;
FIG. 3 is a graph showing the variation in stability of insulin nanoparticles of the present invention;
FIG. 4 is a chart of data table of the pharmacokinetics of buccal patch and sublingual administration of insulin slow release oral patches of the present invention;
Detailed Description
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 disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are described. For purposes of this disclosure, the following terms are defined as follows.
Insulin was produced as bovine insulin by Fumaisi with a bioactivity of 27IU/mg.
Example 1
Preparation of insulin slow-release oral patch
(1) Preparation of polylactic acid-glycolic acid copolymer (PLGA) solution
120mg of polylactic acid-glycolic acid copolymer (50:50) was dissolved in 2ml of methylene chloride to form an organic phase (0.06 g/ml);
(2) Insulin solution preparation
2mg of solid insulin powder was precisely weighed and dissolved in 190. Mu.l of acetate buffer with pH=4 to give a clear solution.
(3) Insulin dispersion solution preparation
190 μl of insulin liquid was uniformly dispersed in 250 μl of sodium alginate solution with a mass-to-volume ratio of 0.003% (w/v) to form an internal aqueous phase.
(4) Preparation of colostrum
Injecting the internal aqueous phase into the organic phase, and performing ultrasonic emulsification under ultrasonic conditions of 10-1000W for 1min to form colostrum (W/O); the volume ratio of the organic phase to the internal water phase is 2:1, a step of;
(5) Preparation of multiple emulsion
Injecting the prepared colostrum into an external water phase containing 10ml of 2% PVA (W/v), 0.045mol/L CaCl2 and 0.00005% (W/v) chitosan, and performing ultrasonic emulsification for 1min under the ultrasonic condition of 10-1000W (ultrasonic condition is the same as above), so as to obtain multiple emulsion (W/O/W); the volume ratio of the colostrum to the external water phase is 1:5, a step of;
(6) Nanoparticle formation
Stirring the compound emulsion at 4deg.C and 1500rpm/min until the organic solvent is completely volatilized.
(7) Freeze drying
Uniformly mixing the insulin nanoparticle solution with 5% mannitol solution, pre-freezing at-20 ℃ for 2 hours, pre-freezing at-80 ℃ for about 24 hours, and freeze-drying in a freeze dryer for 48 hours.
The obtained nano-particle freeze-dried powder is white loose powder, has good redissolution effect, uniform particle dispersion and good stability. FIG. 1 is a scanning electron microscope image of insulin nanoparticles, and FIG. 2 is a distribution range of the particle diameters of the insulin nanoparticles; the graph shows that the obtained insulin nano-particles have round morphology, good fluidity and an average particle diameter of 285nm. The PDI is about 0.2, the encapsulation rate is 60 percent, and the in vitro drug release performance accords with the characteristic of a long-acting preparation.
(8) Preparation of oral Patch
(each tablet weighs about 50mg and contains 4IU of insulin)
Insulin nanoparticle lyophilized powder 1000mg (containing insulin 100 IU)
The freeze-dried powder of the insulin nano-particles is mixed with auxiliary materials such as hypromellose, sodium carboxymethyl cellulose, carbomer, mannitol, magnesium stearate and the like according to the mass ratio of 4:1 for tabletting to obtain tablets with the width of 6 mm and the weight of about 50mg after 2 mm, and the tablets have good pasting performance, good release and moderate dissolution rate.
Example 2
Insulin nanoparticle stability test
The prepared insulin nanoparticles were subjected to PDI (particle distribution density) detection at the same time of day 1-7. The nanoparticle placement condition was 4 ℃, and whether aggregation or degradation occurred was observed. The PDI value is 0.1-0.3 through a An Dongpa LITESIZER 500 particle size analyzer according to the Dynamic Light Scattering (DLS) principle, which shows that the dispersibility is good.
FIG. 3 is a graph showing the variation in stability of insulin nanoparticles of the present invention; from the figure, the insulin nano-particles have good stability at 4 ℃ and no obvious aggregation and degradation phenomenon.
Example 3
Hypoglycemic activity detection of insulin lozenge
The diabetes rat model is induced by the intraperitoneal injection STZ method. The established rat models are randomly divided into A, B, C, D, E, F groups, 6 rats in each group are normally fed for one week, and the rats are fasted for 12 hours before administration.
The tail vein blood was taken after administration for 0, 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, 300, 360, 420, 480min, and blood glucose was measured with a glucometer and blood glucose test paper. Each group was anesthetized and anesthetic was appropriately supplemented during the experiment.
Experimental results as shown in fig. 4, it can be seen from the results that the experimentally prepared (C, D group) insulin nanoparticle patch group has a certain hypoglycemic effect compared with the blank control group. The blood sugar level of rats in the auxiliary material patch group (A) and the blank nano-sheet group (B) fluctuates slightly, but is always at a relatively stable level and does not change obviously, which indicates that auxiliary materials used in the patch preparation process and other reagents in the insulin nano-particle preparation process have no influence on the blood sugar of rats. The (C, D group) insulin nanoparticle patch group gradually reduces the blood sugar of rats, and the insulin nanoparticles are slowly released after being absorbed into blood, so that the effect of reducing the blood sugar is achieved, and a certain slow release effect is achieved.
The insulin slow-release oral patch has the following advantages: 1. the characteristic that insulin can only be injected is changed, and the blood sugar reducing effect can be achieved by oral mucosa blood injection. 2. The acting time of the insulin slow-release oral patch is greatly prolonged to about 10 hours. 3. The dosage is small, each tablet only contains 3-10IU of insulin, and the drug does not have the toxic and side effects of hypoglycemia, insulin resistance and the like caused by high-dose insulin; 4. the oral administration is convenient, the raw materials are safe, the production is convenient, the insulin usage amount is small, the price is relatively low, and the oral administration can be accepted by most patients.

Claims (2)

1. A pharmaceutical formulation characterized in that: the medical auxiliary material comprises nano particles and medical auxiliary materials, wherein the medical auxiliary materials comprise a filler, an adhesive and a lubricant; the dosage form of the pharmaceutical preparation is an oral patch;
the nanoparticle has a bilayer structure, comprising:
(a) A biodegradable polymer inner layer surrounding the solid insulin;
(b) An outer layer having a positive charge;
the biodegradable polymer inner layer component comprises polylactic acid-glycolic acid copolymer, polyvinyl alcohol, sodium alginate and acetate;
the outer layer with positive charge contains chitosan;
the nanoparticle is prepared by the following method:
1) Dissolving solid insulin powder in acetate buffer to form insulin solution;
2) Dispersing insulin solution in sodium alginate solution to serve as an inner water phase, mixing the inner water phase with an organic phase, and performing ultrasonic emulsification to form water-in-oil colostrum;
the external water phase is formed by mixing 0.0001-20.000% w/v polyvinyl alcohol, 0.0001-3.000% w/v chitosan solution and 0.0001-0.18 mol/v Ca2+ solution;
the organic solvent comprises one or more of dichloromethane, ethyl acetate and acetone;
dissolving solid polylactic acid-glycolic acid copolymer in an organic solvent to form an organic phase; the volume ratio of the organic phase to the internal water phase is 2-40:1, the volume ratio of the colostrum to the external water phase is 1:3-20; the mass volume ratio of the sodium alginate solution is 0.0001-50.000% w/v;
3) Injecting the primary emulsion into an external water phase, fully and uniformly mixing, and performing ultrasonic emulsification to form a water-in-oil-in-water compound emulsion solution, wherein the external water phase also contains polyvinyl alcohol, ca2+ and chitosan; wherein the ultrasonic condition is 10-1000w,1-60min ultrasonic;
4) Stirring the re-emulsion solution at a low speed at 4 ℃ or performing vacuum rotary evaporation at room temperature until the organic solvent is completely volatilized; stirring at 1000-5000rpm for 4-12 hr or steaming at 25deg.C for 30-60 min; the freeze drying treatment steps are as follows: adding the nano particles into a mannitol solution containing 5% -7% of m/v, pre-freezing, and then freeze-drying for 48 hours to obtain white powder with uniform texture;
the average particle diameter of the nano particles is 285nm; the PDI is about 0.2;
the filler comprises starch or mannitol, and the adhesive comprises one or more of carbomer, carboxymethyl cellulose and sodium carboxymethyl cellulose; the lubricant comprises magnesium stearate or talcum powder; the mass ratio of the nano particles to the pharmaceutic adjuvant is 4~3: 1-2; is prepared by mixing the nano particles with pharmaceutical excipients and tabletting.
2. Use of a pharmaceutical formulation according to claim 1 for the preparation of a hypoglycemic agent.
CN202111210673.6A 2021-10-18 2021-10-18 Insulin slow-release oral patch and preparation method and application thereof Active CN113893333B (en)

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