CN115998707A - Polydithio polypeptide nanoparticle, preparation method thereof and polypeptide pharmaceutical oral preparation - Google Patents
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Abstract
The invention provides a polydithio polypeptide nanoparticle which is formed by assembling a disulfide heterocyclic compound and a polypeptide drug; the disulfide heterocyclic compound has a structure shown in a formula I. The invention solves the problems of single administration mode of the existing polypeptide drug and defects of other oral polypeptide delivery technologies, and the prepared polydithio polypeptide nano-particles have simple preparation method and convenient use, can overcome three barriers of oral administration, and have higher bioavailability.
Description
Technical Field
The invention relates to the technical field of novel pharmaceutical preparations, in particular to polydithio polypeptide nanoparticles, a preparation method thereof and a polypeptide pharmaceutical oral preparation.
Background
Compared with chemical synthetic medicines, the polypeptide biochemical medicines have the advantages of small side effect, small dosage, good curative effect and the like, and are popular with doctors and patients. At present, a plurality of polypeptide medicaments can be effectively developed through a gene recombination technology, so that the variety of the polypeptide medicaments is increasingly abundant. However, compared with various administration modes of chemical small molecule drugs, the administration mode is relatively single due to low Gastrointestinal (GI) absorption efficiency of polypeptide drugs, and mainly injection administration is adopted.
Among the different modes of non-injectable administration, the oral route of administration is the most traditional, convenient, and easily accepted by patients; compared with injection preparation, the oral administration preparation has stable storage, convenient carrying and use and relatively low cost. If the polypeptide drugs enter the circulatory system to exert the curative effect through oral administration, three physiological barriers need to be overcome: (1) low pH in the stomach and digestive enzymes of the gastrointestinal tract; (2) the mucosal layer of the intestine blocks the diffusion of polypeptide drugs to intestinal epithelial cells; (3) compact intestinal epithelial cells result in extremely low absorption efficiency of polypeptide macromolecules.
Various methods for oral delivery of polypeptides have been reported so far, such as chemical modification of polypeptides, enzyme inhibitors, absorption enhancers, entrapment of polypeptides with carrier materials, and the like. The above strategies have obvious disadvantages, such as covalent modification may reduce the activity of the drug and may change the pharmacological and toxicological properties of the drug molecule; the enzyme inhibitor can effectively prevent the polypeptide medicine from being hydrolyzed by protease, but also affects the digestion of food protein, and can cause digestion and absorption disorder and even pancreatic swelling or hyperplasia after long-term administration; the absorption promoter can increase intestinal permeability and cause irreversible damage to cell membranes, and can cause membrane poisoning after long-term administration; in the process of embedding the polypeptide in the polymer, the participation of an organic solvent is often needed, and steps such as high-speed centrifugation and the like are needed, so that the complexity of the process is increased. Therefore, there is a need to develop a simple, safe, efficient and low cost method of delivering polypeptide drugs.
Disclosure of Invention
In view of the above, the technical problem to be solved by the invention is to provide a polydithio polypeptide nanoparticle, a preparation method thereof and a polypeptide drug oral preparation, wherein the polypeptide drug oral preparation has higher bioavailability when being orally applied.
The invention provides a polydithio polypeptide nanoparticle which is formed by assembling a disulfide heterocyclic compound and a polypeptide drug;
the disulfide heterocyclic compound has a structure shown in a formula I:
X-L-Y formula I;
wherein X is a heterocyclic group containing two or more sulfur atoms;
y is a group that interacts with a polypeptide drug;
l is a X, Y linking group.
The heterocyclic group X is preferably a 4-to 100-membered ring.
Preferably, the X has any one of the following structures:
in the present invention, L is a linking group (linker) between the functional group a and the functional group B.
Preferably, the L is selected from the group consisting of carbon-carbon bonds, carbon-boron bonds, carbon-nitrogen bonds, carbon-phosphorus bonds, carbon-oxygen bonds, carbon-sulfur bonds, carbon-selenium bonds, carbon-tellurium bonds, metal coordination bonds, boron ester bonds, disulfide bonds, cyclic groups, hydrogen bonds, cleavable chemical bonds, supramolecular host guest interactions, or ligand-receptor recognition interactions.
The above-mentioned groups are L and X bonding mode or L and Y bonding mode. The bonding between L and X and the bonding between L and Y may be the same or different.
The mode of action of the above-mentioned Y and polypeptide drugs may be alone or in combination include covalent and non-covalent actions.
Preferably, the Y is selected from one or more of chemical/biological molecules recognizing polypeptide drugs, DNA complementary strands, aptamers, polypeptides forming a coded structures/zippers/superstructures, and the following chemical groups:
preferably, the disulfide heterocyclic compound has a structure shown in a formula II:
wherein X is 1 、X 2 、X 3 、X 4 At least two of which are S, and the rest are C;
R 1 、R 2 、R 3 independently selected from carbon atoms, amide groups or imino groups;
n1, n2, n3, n4 are independently selected from any integer from 1 to 6.
In the above structure of the invention, the curved lineRepresenting the connection location; the single bond "-" represents methyl.
Single bonds to cyclic or aromatic ring groups, e.g.And represents any position where a methyl group or an ethyl group may be attached to a cyclic group or an aromatic cyclic group, respectively.
Substituents for cyclic or aromatic radicals, e.g.Respectively, represents that the substituent may be attached to any position of the cyclic group or the aromatic ring group.
Preferably, the disulfide heterocyclic compound has one or more of the following structures:
the disulfide heterocyclic compound monomer is connected to the polypeptide medicine due to the existence of the Y group, so that the disulfide heterocyclic compound is gathered on the surface of the polypeptide medicine, sulfur-containing heterocycle is enriched, concentration-driven ring-opening polymerization reaction is carried out to form a polydisulfide compound, and the polypeptide medicine is wrapped to form nanoparticles.
In the invention, the number of the disulfide heterocyclic compounds in the polypeptide drug and the disulfide heterocyclic compound nanoparticle can be one, two or more.
In the invention, polypeptide drug particles are taken as template molecules, and disulfide molecules are taken as basic encapsulation molecules. One end of the disulfide compound is disulfide heterocycle, and the other end is a group combined with polypeptide medicine. The polypeptide drug and the disulfide compound are self-assembled into nano particles, and ring-opening polymerization reaction is carried out on the heterocyclic ends of the disulfide compound on the surfaces of the nano particles, so that the polypeptide nano particles wrapped by the polydisulfide compound are formed. The polypeptide nanoparticle is stable in the gastrointestinal tract environment, and enters the circulatory system through the intestinal epithelial cells by virtue of dynamic chemical exchange reaction between the polydisulfide molecular bond on the surface and the intestinal mucin in intestinal juice and the sulfhydryl of the inner and outer proteins of the epithelial cell membranes. The polypeptide nano particles are dispersed in aqueous or non-aqueous liquid to form various types of polypeptide medicine oral liquid, and the polypeptide nano particles are mixed with auxiliary materials to form polypeptide medicine capsules, tablets, spray and the like.
The type of the polypeptide drug is not particularly limited, and is preferably one or more of glucose oxidase, coagulation factor VIII, neurotensin, haemagglutinin, melanocyte stimulating hormone, thymosin, thymopoietin, corticotropin, trypsin inhibitor, chorionic gonadotrophin, protamine, human gamma globulin, albumin, gastric membranogen, epidermal growth factor, erythropoietin, interleukin-2, cytokine-type drug, and the like.
Preferably, the glucose oxidase is selected from one or more of growth hormone, calcitonin, vancomycin, polymyxin, interferon, hirudin, cyclosporin (cyclopeptide) and erythropoietin.
Preferably, the molar ratio of the dithioheterocyclic compound to the polypeptide drug is 1-200:1.
In the invention, the particle size of the polydithio polypeptide nano-particles is preferably 20-200 nm.
In the invention, the polypeptide drug and the disulfide heterocyclic compound nanoparticle have high stability in simulated gastric and intestinal fluids, and can avoid degradation and leakage of the polypeptide drug in an acidic environment and a digestive enzyme environment.
The invention provides a preparation method of the polydithio polypeptide nanoparticle, which comprises the following steps:
s1) mixing and incubating polypeptide drug solution and dithioheterocyclic compound solution, and assembling to form nano particles.
Specifically, firstly, mixing and incubating polypeptide drug solution and disulfide heterocyclic compound solution, and assembling to form nano particles; and then removing free disulfide heterocyclic compounds and/or polypeptide drugs which do not participate in assembly to form nano particles, so as to obtain a suspension of polydisulfide polypeptide nano particles.
The time of the mixed incubation is preferably 30min.
The method of removing the free dithioheterocyclic compound and/or polypeptide drug which does not participate in the assembly to form nanoparticles is not particularly limited in the present invention, and may be a method well known to those skilled in the art, including but not limited to dialysis or centrifugation.
In the present invention, the above-mentioned assembly is preferably carried out in a neutral aqueous solution. Wherein, the polypeptide drug particles dissolved in water are used as template molecules, and disulfide molecules are used as basic encapsulation molecules. The disulfide compound spontaneously forms a polymer on the surface of the polypeptide drug, and the polypeptide drug is wrapped to form nanoparticles.
The preparation method disclosed by the invention has mild conditions and simple method.
The invention discloses application of the polydithio polypeptide nano-particles in preparation of polypeptide medicaments.
In the present invention, the aqueous suspension of polydithio polypeptide nanoparticles may be directly administered orally to mammals.
Based on the above, the invention provides a polypeptide pharmaceutical oral preparation, which comprises the polydithio polypeptide nanoparticle and pharmaceutically acceptable auxiliary agents.
The dosage form of the oral preparation of the present invention is not particularly limited, and may be an oral dosage form well known to those skilled in the art.
Preferably, the oral preparation is one or more of enteric coated capsules, powder, tablets, granules, suspension, dripping pills, oral liquid and spray.
The polydithio polypeptide nano-particles are dispersed in aqueous or non-aqueous liquid to form various types of polypeptide medicine oral liquid.
Mixing the polydithio polypeptide nanoparticle with pharmaceutically acceptable auxiliary materials to form oral preparations such as polypeptide medicine capsules, powder, tablets, granules, suspension, dripping pills and the like.
The kind of the auxiliary agent is not particularly limited in the present invention, and a person skilled in the art may select it according to the dosage form.
In the invention, the polypeptide drug oral preparation also comprises other therapeutic agents;
the other therapeutic agent is selected from subcutaneous injection type polypeptide medicine and/or other oral type polypeptide medicine.
Compared with the prior art, the invention provides a polydithio polypeptide nanoparticle which is formed by assembling a disulfide heterocyclic compound and a polypeptide drug; the disulfide heterocyclic compound has a structure shown in a formula I.
The invention solves the problems of single administration mode of the existing polypeptide drug and defects of other oral polypeptide delivery technologies, and the prepared polydithio polypeptide nano-particles have simple preparation method and convenient use, can overcome three barriers of oral administration, and have higher bioavailability.
Drawings
FIG. 1 is a nuclear magnetic resonance characterization of compound 1 of example 1;
FIG. 2 is a TEM profile of growth hormone-loaded nanoparticles of example 1; ruler: 100nm;
FIG. 3 is a graph showing the release profile of the growth hormone-loaded nanoparticles of example 1 under various conditions;
FIG. 4 is a graph showing the time profile of the growth hormone-loaded nanoparticles in example 1 across cells;
FIG. 5 is a graph showing the blood concentration of hollow intestine injected growth hormone nanoparticles of example 1;
FIG. 6 is a graph showing the blood concentration of lyophilized powder of the capsule-administered growth hormone nanoparticles of example 1;
FIG. 7 is a TEM profile of salmon-carried calcitonin nanoparticles in example 2; ruler: 100nm;
FIG. 8 is a graph showing the release profile of salmon calcitonin nanoparticles in example 2 under various conditions;
FIG. 9 is a graph of the cell-crossing time profile of salmon calcitonin nanoparticles in example 2;
FIG. 10 is a TEM morphology of vancomycin-loaded nanoparticles of example 3; ruler: 100nm;
FIG. 11 is a release profile of vancomycin-loaded nanoparticles of example 3 under various conditions;
FIG. 12 is a graph showing the transcellular time profile of vancomycin-loaded nanoparticles of example 3.
Detailed Description
In order to further illustrate the present invention, the polydithio polypeptide nanoparticles, the preparation method thereof and the polypeptide pharmaceutical oral preparation provided by the present invention are described in detail below with reference to examples.
Example 1
1.1 preparation of the Dithiaheterocyclic Compounds
This example employs compound 1 of the formula:
the synthesis of the compound 1 in the invention comprises the following steps:
dissolving the compound 2 and N, N' -carbonyl diimidazole in dichloromethane, dropwise adding the solution into a dichloromethane solution of ethylenediamine under ice water bath conditions, stirring for 40 minutes, and stirring for 30 minutes at room temperature; the reacted solution was washed with brine, and the organic phase portion was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to give a yellow oil, to give compound 3. Dissolving the compound 3 in dichloromethane, adding 1H-pyrazole-1-formamidine hydrochloride, stirring at room temperature for 4 hours, distilling under reduced pressure to remove the solvent, dissolving the residue in methanol, adding diethyl ether to induce precipitation, collecting the solid, and washing with diethyl ether to obtain pale yellow solid, namely the compound 1 in the experiment.
The structure of the product was characterized by nuclear magnetic resonance and the results are shown in figure 1.
1.2 preparation of growth hormone nanoparticles
The recombinant human growth hormone and the compound 1 are mixed, stirred and incubated for 30 minutes in a buffer solution with the pH value of 7.4 and the Tris-HCl of 20mM according to the mass ratio of 1:50, and the stirring speed is 300rpm, so that nanoparticle suspension is obtained. And (3) dialyzing or centrifuging the obtained suspension to remove unreacted free growth hormone and free compound 1, thereby obtaining the assembled nanoparticle of the growth hormone and the dithioheterocyclic compound 1. The average particle diameter was 40nm. The morphology of the growth hormone nanoparticle is shown in figure 2.
1.3 determination of the drug-loading and encapsulation Rate of growth hormone-loaded nanoparticles
Drug loading and encapsulation efficiency are commonly used to represent the drug loading capacity of nanoparticles, and the encapsulation efficiency and drug loading are calculated by measuring free growth hormone in the supernatant of the nanoparticles subjected to dialysis by using high performance liquid chromatography. And measuring the content of growth hormone in the supernatant, and further calculating the drug loading rate and the encapsulation efficiency. Wherein, the drug loading rate refers to the ratio of the drug loading rate of the package entering the nanoparticle to the total weight (carrier+packaged drug); encapsulation efficiency refers to the ratio of the amount of drug encapsulated into the nanoparticle to the amount administered. The calculation formulas of the encapsulation efficiency and the drug loading rate are respectively as follows:
wherein M is Total (S) For initial growth hormone mass (mg), V External cleaner Volume of supernatant (mL), C External cleaner Is the concentration of growth hormone (mg/mL) in the supernatant, W Nanoparticles The mass (mg) of the obtained nanoparticle is shown.
The medicine carrying mass percent of the measured growth hormone nanoparticle is 42.3%, and the encapsulation efficiency is 69.6%.
1.4 study of the release kinetics of growth hormone nanoparticles under different conditions
As in vitro simulated gastric fluid and simulated intestinal fluid, a hydrochloride buffer solution with ph=1.2 and a phosphate buffer solution with ph=7.4 were used, respectively. The release behavior of growth hormone was analyzed in 7mM solutions of simulated gastric fluid, simulated intestinal fluid and glutathione.
1.0mg of FITC-labeled growth hormone-carrying nanoparticles were accurately weighed and dissolved in 7mM physiological buffer solutions simulating gastric juice, intestinal juice and glutathione, respectively, and the solutions were added to a microdialysis device, respectively, and released at 37 ℃. At the indicated time points, 50 μl of solution was removed from the supernatant and supplemented with an equivalent amount of simulated gastric fluid, intestinal fluid or glutathione solution. 50. Mu.L of the solution was diluted to 200. Mu.L, and the fluorescence intensity was measured with a fluorescence spectrophotometer, and the amount of released growth hormone at each time point was calculated by a standard curve.
As shown in fig. 3, the growth hormone nanoparticles are slowly released in simulated gastric fluid, and the accumulated growth hormone release amount is less than 10% in 12 hours; hardly released in simulated intestinal fluid; growth hormone was rapidly released in 7mM glutathione physiological buffer. Furthermore, circular dichroism characterization found that the secondary structure remained intact after growth hormone release, which is one of the markers of growth hormone bioactivity. The experimental result shows that the growth hormone nano-particles can be kept stable in the gastrointestinal tract environment.
1.5 transcellular study of growth hormone nanoparticles in vitro intestinal models
Inoculating Caco-2 cells into 24-well Transwell culture chamber, culturing for 21 days, and detecting transepithelial resistance (TEER) of cell monolayer with Millicell resistance meter to more than 300Ω cm 2 The tight connection between Caco-2 cells is shown, and the in vitro intestinal epithelial cell monolayer model is successfully constructed.
100. Mu.L of 20. Mu.g/mL nanoparticle suspension was added to the Transwell upper chamber, and 100. Mu.L of liquid was withdrawn from the substrate side chamber at the time points of 0, 0.25h, 0.5h, and 1h in the previous hour, and then samples were taken every one hour until 12h. Immediately after each sample, the extracted sample volume was replaced with an equal amount of experimental medium. TEER measurements were also performed at the same time point to determine the integrity of the single layer epithelial cells. 100. Mu.L of a 9. Mu.g/mL growth hormone solution was added to the above chamber as a negative control.
As shown in FIG. 4, the efficiency of transferring growth hormone across epithelial cells by the growth hormone-carrying nanoparticles was about 69%, whereas little growth hormone penetrated the epithelial cells in the control group. During the experiment, TEER values of the nanoparticle sets did not change significantly, indicating that nanoparticle transport across cells did not disrupt the tight junctions between cells.
1.6 study of blood concentration changes after jejunal injection of the rat with the suspension of growth hormone nanoparticles
Adult male Sprague-Dawley (SD) rats were used for the experiment, with weights of 275-300 g. All animals were fasted for 7h prior to the experiment. Rats were anesthetized with 1.5% -3.0% isoflurane prior to surgery. The gastrointestinal system is exposed by cutting 2.5-3.0 cm in the middle of abdomen. 3mg/kg of the suspension of growth hormone nanoparticles is injected at a distance of about 10-15 cm from the proximal end of the small intestine. After injection, the injection hole is sealed with a minimum amount of surgical glue. The midline muscle incision is then closed with sterile surgical sutures, and the skin incision is then closed with a wound clip. Then 50. Mu.L of blood was collected from each orbit at predetermined time intervals, the blood sample was centrifuged at 5000rpm for 10min, and the upper plasma was collected and stored at-80℃for analysis. The human growth hormone kit was used to determine the concentration of growth hormone in blood samples, and the results are shown in FIG. 5.
1.7 study of blood concentration changes of nanoparticle lyophilized powder coated with oral enteric-coated capsule
The in vivo effect of oral administration of the formulation was verified by performing a gastric lavage experiment with adult male SD rats (300-350 g). Animals were fasted for 7h prior to the experiment. The administration was performed by intragastric administration using a hard gelatin capsule No. 9, which contained lyophilized powder of growth hormone nanoparticles at a dose of 3mg/kg. No. 9 capsules were purchased from Torpac, inc. of USA and coated with 10% w/v of Eudragit L30D-55 solution.
After the capsule was administered to the rats with the drug dispenser, 0.5 ml of sterile physiological saline was filled in the stomach. Blood samples were then collected at predetermined time points, 50 μl of each blood sample was collected, centrifuged at 5000rpm for 10min, and the upper plasma was stored at-80 ℃ for analysis. The human growth hormone kit was used to determine the concentration of growth hormone in blood samples, and the results are shown in FIG. 6.
Example 2
2.1 preparation of Dithiaheterocyclic Compounds 4
The dithioheterocyclic compounds used in this example are:
the synthesis steps are as follows
Compound 5 was dissolved in 4mol/L aqueous sodium hydroxide solution, compound 6 was dissolved in 4mol/L aqueous sodium hydroxide solution, and the two solutions were mixed. Mixing and stirring for 2h at 70 ℃, and regulating the pH value of the reaction mixture to be 9-10 in the whole reaction time. At the end of the reaction, the mixture was heated at 90℃for about 10min, and after cooling the reaction mixture to room temperature, concentrated hydrochloric acid was added. Concentrating the solution in a vacuum evaporator as much as possible, adding absolute ethanol into the concentrated oil, separating insoluble crystals, filtering, collecting, and washing with warm ethanol. The crystals obtained were a mixture of the sodium salt of compound 4 with sodium chloride. It was recrystallized from 2mol/L hydrochloric acid and then from 0.2mol/L hydrochloric acid. Compound 4 was obtained.
2.2 preparation of calcitonin nanoparticles
The remainder of the procedure was the same except that part 1.2 of the growth hormone in example 1 was changed to salmon calcitonin, and compound 1 was changed to compound 4. The average particle diameter was 30nm. The calcitonin nanoparticle morphology is shown in figure 7.
2.3 determination of drug-loading and encapsulation efficiency of salmon calcitonin nanoparticle
The procedure is as in example 1, section 1.3.
The drug loading mass percentage of the measured salmon calcitonin nanoparticle is 34.6%, and the encapsulation efficiency is 72.3%.
2.4 salmon calcitonin nanoparticle release kinetics study under different conditions
The procedure is as in section 1.4 of example 1.
The experimental results are shown in FIG. 8 and are similar to those of 1.4.
2.5 cell-crossing study of salmon calcitonin nanoparticle in an in vitro intestinal model
The procedure is as in example 1, section 1.5.
As shown in FIG. 9, the efficiency of transfer of salmon calcitonin from the salmon calcitonin-carrying nanoparticle across the epithelial cells was about-49%, whereas almost no salmon calcitonin penetrated the epithelial cells in the control group. During the experiment, TEER values of the nanoparticle sets did not change significantly, indicating that nanoparticle transport across cells did not disrupt the tight junctions between cells.
Example 3
3.1 preparation of dithioheterocyclic Compound 7
The dithioheterocyclic compounds used in this example are:
the synthesis steps are as follows
3.2 preparation of vancomycin nanoparticles
The growth hormone of example 1, part 1.2, was changed to vancomycin, compound 1 was changed to compound 7, the ratio of the amounts of vancomycin and compound 7 was 1:30, and the other steps were the same. The average particle diameter was 35nm. The morphology of vancomycin nanoparticles is shown in figure 10.
3.3 determination of drug-loading and encapsulation Rate of vancomycin nanoparticles
The procedure is as in example 1, section 1.3.
The drug loading mass percentage of the measured vancomycin nano-particles is 34.1%, and the encapsulation efficiency is 71.8%.
3.4 release kinetics study of vancomycin nanoparticles under different conditions
The procedure is as in section 1.4 of example 1.
The experimental results are shown in FIG. 11 and are similar to those of 1.4.
3.5 transcellular study of vancomycin nanoparticles in an in vitro intestinal model
The procedure is as in example 1, section 1.5.
As shown in FIG. 12, the efficiency of transferring vancomycin across epithelial cells by vancomycin-loaded nanoparticles was about 60%, whereas almost no vancomycin penetrated the epithelial cells in the control group. During the experiment, TEER values of the nanoparticle sets did not change significantly, indicating that nanoparticle transport across cells did not disrupt the tight junctions between cells.
The above description of the embodiments is only for aiding in the understanding of the method of the present invention and its core ideas. It should be noted that it will be apparent to those skilled in the art that various modifications and adaptations of the invention can be made without departing from the principles of the invention and these modifications and adaptations are intended to be within the scope of the invention as defined in the following claims.
Claims (13)
1. A polydithio polypeptide nanoparticle, which is a nanoparticle formed by assembling a disulfide heterocyclic compound and a polypeptide drug;
the disulfide heterocyclic compound has a structure shown in a formula I:
X-L-Y is of formula I;
wherein X is a heterocyclic group containing two or more sulfur atoms;
y is a group that interacts with a polypeptide drug;
l is a X, Y linking group.
2. The polydithio polypeptide nanoparticle of claim 1 wherein X has any of the following structures:
the L is selected from a carbon-carbon bond, a carbon-boron bond, a carbon-nitrogen bond, a carbon-phosphorus bond, a carbon-oxygen bond, a carbon-sulfur bond, a carbon-selenium bond, a carbon-tellurium bond, a metal coordination bond, a boron ester bond, a disulfide bond, a cyclic group, a hydrogen bond, a cleavable chemical bond, a supermolecule host-guest effect or a ligand-receptor recognition effect;
the Y is selected from one or more of a chemical/biological molecule recognizing polypeptide drugs, a DNA complementary strand, an aptamer, a polypeptide forming a conjugated structures/zipping structures/superstructures, and the following chemical groups:
3. the polydithio polypeptide nanoparticle of claim 1 wherein the dithioheterocyclic compound has a structure of formula ii:
wherein X is 1 、X 2 、X 3 、X 4 At least two of which are S, and the rest are C;
R 1 、R 2 、R 3 independently selected from carbon atoms, amide groups or imino groups;
n1, n2, n3, n4 are independently selected from any integer from 1 to 6.
5. the polydithio polypeptide nanoparticles of claim 1 wherein the polypeptide drug is selected from one or more of glucose oxidase, blood coagulation factor viii, neurotensin, haemagglutinin, melanocyte stimulating hormone, thymosin, thymopoietin, corticotropin, trypsin inhibitor, chorionic gonadotrophin, protamine, human gamma globulin, albumin, gastric membranogen, epidermal growth factor, erythropoietin, interleukin-2, cytokine class drugs.
6. The polydithio polypeptide nanoparticles of claim 5 wherein the glucose oxidase is selected from one or more of the group consisting of growth hormone, calcitonin, vancomycin, polymyxin, interferon, hirudin, cyclosporin, and erythropoietin.
7. The polydithio polypeptide nanoparticle of claim 1 wherein the molar ratio of disulfide heterocyclic compound to polypeptide drug is 1-200:1.
8. The polydithio polypeptide nanoparticle of claim 1 wherein the polydithio polypeptide nanoparticle has a particle size of 20-200 nm.
9. The method for preparing polydithio polypeptide nanoparticles of any one of claims 1 to 8, comprising the steps of:
s1) mixing and incubating polypeptide drug solution and dithioheterocyclic compound solution, and assembling to form nano particles.
10. Use of the polydithio polypeptide nanoparticles of any one of claims 1 to 8 for the preparation of polypeptide pharmaceuticals.
11. A polypeptide pharmaceutical oral formulation comprising the polydithio polypeptide nanoparticle of any one of claims 1-8 and a pharmaceutically acceptable adjuvant.
12. The polypeptide pharmaceutical oral preparation according to claim 11, wherein the oral preparation is one or more of enteric coated capsules, powders, tablets, granules, suspensions, drop pills, oral liquids and sprays.
13. The polypeptide pharmaceutical oral formulation of claim 11, further comprising an additional therapeutic agent;
the other therapeutic agent is selected from subcutaneous injection type polypeptide medicine and/or other oral type polypeptide medicine.
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CN114404607A (en) * | 2019-03-08 | 2022-04-29 | 苏州维益生物科技有限公司 | Nucleic acid spherical nano-particle medicine, preparation method and application thereof |
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