Pharmaceutical composition containing bioactive substances as well as preparation method and application of pharmaceutical composition

Abstract

The invention relates to a pharmaceutical composition containing bioactive substances as well as a preparation method and application of the pharmaceutical composition. The pharmaceutical composition comprises a bioactive substance, a polymer and a penetration enhancer, the bioactive substance is selected from the group consisting of tirpotide, semaglutide or liraglutide; the polymer comprises a framework type polymer and an enteric polymer. The pharmaceutical composition has good permeability in intestinal juice and can expand in the gastric juice to form a gel skeleton after oral administration, the polypeptide drug in the pharmaceutical composition can be kept stable in the stomach, and the polypeptide drug and a penetration enhancer are quickly released after entering the small intestine, so that the absorption rate of polypeptide in the intestinal tract is greatly improved, and the bioavailability is improved. Therefore, the bioavailability of oral administration of the polypeptide medicine is improved.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical preparations, and in particular relates to a pharmaceutical composition containing a bioactive substance, and a preparation method and application thereof. Background Art

[0002] Among biological preparations, peptides are particularly favored due to their excellent specificity, selectivity, safety and efficacy. Currently, there are more than 80 peptide drugs approved worldwide, more than 90% of which are injectable preparations, and only 4% are administered orally. The oral administration of peptide drugs is limited by local conditions in the gastrointestinal tract. Due to susceptibility to pH and gastrointestinal enzymes, as well as low intestinal epithelial cell membrane permeability, oral administration of hydrophilic macromolecules with a relative molecular mass of more than 1000Da remains a challenge.

[0003] Currently available peptide products use various delivery technologies, but the oral bioavailability of hydrophilic peptides is still low. (hydrophilic peptide), using solid self-nanoemulsifying drug delivery system (SNEDDS) technology, the oral bioavailability is only 0.17%; semaglutide (hydrophilic peptide) add absorption enhancer, use Carrier technology, oral bioavailability is only 0.4% to 1%; octreotide The use of absorption enhancers and enteric coating technology has resulted in an oral bioavailability of approximately 0.25%; Voclosporin (trade name ) The structure of CsA was optimized (to increase the lipid solubility of the polypeptide) and SNEDDS technology (lipid drug delivery system) was used at the same time, with an oral bioavailability of about 8%.

[0004] Therefore, developing a pharmaceutical composition containing bioactive substances to improve the absorption of polypeptide bioactive substances by intestinal epithelial cells and improve bioavailability has become a technical problem that needs to be solved urgently. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention aims to provide a pharmaceutical composition containing a bioactive substance, a preparation method thereof and an application thereof. The pharmaceutical composition has good permeability in intestinal fluid, and after oral administration, it can swell in gastric fluid to form a gel skeleton, and the polypeptide drug therein can remain stable in the stomach, and after entering the small intestine, the polypeptide drug and the permeation enhancer are quickly released, which greatly improves the absorption rate of the polypeptide in the intestine, thereby improving the bioavailability of the polypeptide drug after oral administration.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a pharmaceutical composition comprising a bioactive substance, the pharmaceutical composition comprising a bioactive substance, a polymer and a penetration enhancer;

[0008] The bioactive substance is selected from telpotide, semaglutide or liraglutide.

[0009] The polymers include backbone polymers and enteric polymers.

[0010] In the present invention, the skeleton-type polymer does not dissolve after encountering gastric acid in the stomach and can quickly form a gel skeleton, which can reduce the diffusion of bioactive substances and permeation enhancers in the stomach.

[0011] Enteric polymers are insoluble in gastric juice (pH around 1.0) and block gastric juice from entering the composition, further reducing the diffusion of active substances and penetration enhancers in the stomach. Enteric polymers dissolve rapidly in intestinal juice (pH above 5.5) and can promote the rapid diffusion of bioactive substances and penetration enhancers in the small intestine.

[0012] Permeation enhancers can act on biological membranes and change the tight junctions of the gastrointestinal epithelium, thereby improving the permeability of drugs. After the drug composition enters the intestine, the permeation enhancer is released to change the pH value of the intestinal environment, thereby protecting the active substances by inhibiting the activity of intestinal proteases and improving the absorption of active substances by acting on epithelial cells.

[0013] The bioactive substance in the present invention is selected from telpotide (relative molecular mass 4813.45Da), semaglutide (relative molecular mass 4113.58Da) or liraglutide (relative molecular mass 3751.20Da). Under the combined action of the backbone polymer, the enteric polymer and the permeation enhancer, the above-mentioned macromolecular polypeptide improves its absorption capacity in the intestine and further improves the bioavailability.

[0014] Preferably, the pharmaceutical composition comprises, by mass percentage, 5-30% of biologically active substance, 55-70% of polymer and 15-30% of penetration enhancer.

[0015] Preferably, the mass percentage of the backbone polymer in the polymer is 45-55%.

[0016] Among them, the specific point values ​​in 5-30% can be selected from 5%, 10%, 15%, 20%, 25%, 30% etc., the specific point values ​​in 55-70% can be selected from 55%, 58%, 61%, 64%, 67%, 70% etc., the specific point values ​​in 15-30% can be selected from 15%, 18%, 21%, 24%, 27%, 30% etc., and the specific point values ​​in 45-55% can be selected from 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55% etc.

[0017] Based on the combined effects of the backbone polymer, the enteric polymer and the penetration enhancer in improving the intestinal absorption capacity of the bioactive substance, when the components satisfy the above-mentioned mass percentage relationship, the bioavailability of the active substance can be better improved.

[0018] Preferably, the bioactive substance is selected from telportopide; the backbone polymer includes hyaluronic acid and carboxymethyl chitosan; the enteric polymer includes hydroxypropyl methylcellulose acetate succinate, methacrylic acid-ethyl acrylate copolymer and acrylic resin; the penetration enhancer includes capric acid, chenodeoxycholic acid and 8-(2-hydroxybenzamido)octanoic acid.

[0019] In the present invention, when the bioactive substance is telpotide, the above-mentioned specific backbone polymer can improve the stability of the bioactive substance and synergistically reduce the dissolution of telpotide in gastric juice; the above-mentioned specific enteric polymer can maintain the rapid release of telpotide in the intestine; and the above-mentioned specific penetration enhancer can further improve the permeability of telpotide in the intestine.

[0020] Preferably, the mass ratio of the hyaluronic acid to carboxymethyl chitosan is 1:(0.1-1).

[0021] Preferably, the mass ratio of the hydroxypropyl methylcellulose acetate succinate, the methacrylic acid-ethyl acrylate copolymer and the acrylic resin is 1:(0.1-1):(0.1-1).

[0022] Preferably, the mass ratio of decanoic acid, chenodeoxycholic acid and 8-(2-hydroxybenzamido)octanoic acid is 1:(0.1-1):(0.1-1).

[0023] Among them, the specific point values ​​between 0.1 and 1 can be selected from 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, etc.

[0024] Preferably, the bioactive substance is selected from semaglutide; the skeleton polymer includes polylactic acid and hydroxypropyl chitosan; the enteric polymer includes hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate and acrylic resin; the penetration enhancer includes 8-(2-hydroxybenzamido)octanoic acid, ethylenediaminetetraacetic acid and bile acid.

[0025] In the present invention, when the bioactive substance is semaglutide, the above-mentioned specific backbone polymer can improve the stability of the bioactive substance and synergistically reduce the dissolution of semaglutide in gastric juice; the above-mentioned specific enteric polymer can maintain the rapid release of semaglutide in the intestine; the above-mentioned specific penetration enhancer can further improve the permeability of semaglutide in the intestine.

[0026] Preferably, the mass ratio of the polylactic acid to hydroxypropyl chitosan is 1:(0.1-1).

[0027] Preferably, the mass ratio of hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate and acrylic resin is 1:(0.1-1):(0.1-1).

[0028] Preferably, the mass ratio of 8-(2-hydroxybenzamido)octanoic acid, ethylenediaminetetraacetic acid and cholic acid is 1:(0.1-1):(0.1-1).

[0029] Among them, the specific point values ​​between 0.1 and 1 can be selected from 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, etc.

[0030] Preferably, the bioactive substance is selected from liraglutide; the backbone polymer includes alginate and sodium hyaluronate; the enteric polymer includes methacrylic acid-ethyl acrylate copolymer, cellulose acetate phthalate and hydroxypropyl methylcellulose phthalate; and the penetration enhancer includes caprylic acid, capric acid and citric acid.

[0031] In the present invention, when the bioactive substance is liraglutide, the above-mentioned specific backbone polymer can improve the stability of the bioactive substance and synergistically reduce the dissolution of liraglutide in gastric juice; the above-mentioned specific enteric polymer can maintain the rapid release of liraglutide in the intestine; and the above-mentioned specific penetration enhancer can further improve the permeability of liraglutide in the intestine.

[0032] Preferably, the mass ratio of alginate to sodium hyaluronate is 1:(0.1-1).

[0033] Preferably, the mass ratio of the methacrylic acid-ethyl acrylate copolymer, cellulose acetate phthalate and hydroxypropyl methylcellulose phthalate is 1:(0.1-1):(0.1-1).

[0034] Preferably, the mass ratio of caprylic acid, capric acid and citric acid is 1:(0.1-1):(0.1-1).

[0035] Among them, the specific point values ​​between 0.1 and 1 can be selected from 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, etc.

[0036] In a second aspect, the present invention provides a method for preparing a pharmaceutical composition comprising a bioactive substance as described in the first aspect, the method comprising the following steps:

[0037] (1) dissolving the bioactive substance, the enteric polymer and the permeation enhancer in solvent I to obtain solution A; dissolving the backbone polymer in solvent III to obtain solution B;

[0038] (2) Solution A and solution B are mixed and extruded to obtain a pharmaceutical composition solution, which is then dried to obtain a pharmaceutical composition containing a bioactive substance.

[0039] Preferably, the solvent I is an alkaline solution, preferably an aqueous sodium hydroxide solution.

[0040] Preferably, the solvent II includes any one of methanol, ethanol, isopropanol or acetone, or a combination of at least two of them.

[0041] Preferably, the mixing in step (2) includes mixing using microfluidization technology.

[0042] In the present invention, the microfluidization technology is used to prepare the pharmaceutical composition containing the bioactive substance, which can better mix the components in the composition and improve the stability of the prepared pharmaceutical composition.

[0043] In a third aspect, the present invention provides an oral pharmaceutical preparation comprising a bioactive substance, wherein the pharmaceutical preparation comprises the pharmaceutical composition comprising a bioactive substance as described in the first aspect, and a pharmaceutically acceptable excipient.

[0044] Preferably, in the oral pharmaceutical preparation, the mass percentage of the pharmaceutical composition is 5-70%, for example, it can be 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, etc.

[0045] Preferably, the excipient comprises any one of lactose, magnesium stearate, mannitol or microcrystalline cellulose, or a combination of at least two thereof.

[0046] Preferably, the dosage form of the pharmaceutical preparation is a solid or semisolid preparation.

[0047] Preferably, the solid preparation includes tablets, granules or capsules.

[0048] Preferably, the semisolid preparation comprises a suspension or an emulsion.

[0049] The numerical range described in the present invention not only includes the point values ​​listed above, but also includes any point values ​​between the above numerical ranges that are not listed. Due to space limitations and for the sake of simplicity, the present invention no longer exhaustively lists the specific point values ​​included in the range.

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] The pharmaceutical composition developed by the present invention has good in vitro permeability. After oral administration, it can swell in gastric juice to form a gel skeleton, and the polypeptide drug therein can remain stable in the stomach. After entering the small intestine, the polypeptide drug and the permeation enhancer are quickly released, which greatly improves the absorption rate of the polypeptide in the intestine, thereby improving the bioavailability of the polypeptide drug after oral administration. DETAILED DESCRIPTION

[0052] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0053] The sources of the components / raw materials in the following embodiments are shown in Table 1:

[0054] Table 1

[0055]

[0056]

[0057] Example 1: Pharmaceutical composition comprising telpotide

[0058] The present embodiment provides a pharmaceutical composition comprising tilpoitide, and the formula is as follows: polypeptide (tilpoitide 20 mg), backbone polymer 60 mg (hyaluronic acid 30 mg, carboxymethyl chitosan 30 mg), enteric polymer 60 mg (hydroxypropyl methylcellulose acetate succinate 20 mg, methacrylic acid-ethyl acrylate copolymer 20 mg, acrylic resin 20 mg), penetration enhancer 60 mg (decanoic acid 20 mg, chenodeoxycholic acid 20 mg, 8-(2-hydroxybenzamido) caprylic acid 20 mg).

[0059] Preparation process:

[0060] (1) dissolving the polypeptide in a sodium hydroxide aqueous solution and adjusting the pH of the solution to 7.0, and dissolving the permeation enhancer and the enteric polymer in a sodium hydroxide aqueous solution and adjusting the pH of the solution to 7.0;

[0061] (2) Mixing the two solutions using a high-speed shearing machine for 30 minutes until uniformly mixed, to obtain solution A;

[0062] (3) dissolving the backbone polymer in 300 times the volume of ethanol solution to obtain solution B;

[0063] (4) Solution A and Solution B were mixed by microfluidization technology, with the speed ratio of Solution A to Solution B being 2:3, and were repeatedly extruded by a 200 nm polycarbonate membrane extruder, and the organic solvent was removed by dialysis, and then freeze-dried to obtain a pharmaceutical composition containing telpotide.

[0064] Example 2: Pharmaceutical composition comprising telpotide

[0065] The present embodiment provides a pharmaceutical composition comprising tilpoitide, and the formula is as follows: polypeptide (tilpoitide 20 mg), backbone polymer 60 mg (hyaluronic acid 40 mg, carboxymethyl chitosan 20 mg), enteric polymer 60 mg (hydroxypropyl methylcellulose acetate succinate 15 mg, methacrylic acid-ethyl acrylate copolymer 30 mg, acrylic resin 15 mg), penetration enhancer 60 mg (decanoic acid 30 mg, chenodeoxycholic acid 10 mg, 8-(2-hydroxybenzamido) caprylic acid 20 mg).

[0066] Preparation process:

[0067] (1) dissolving the polypeptide in a sodium hydroxide aqueous solution and adjusting the pH of the solution to 7.0, and dissolving the permeation enhancer and the enteric polymer in a sodium hydroxide aqueous solution and adjusting the pH of the solution to 7.0;

[0068] (2) Mixing the two solutions using a high-speed shearing machine for 40 minutes until uniformly mixed, to obtain solution A;

[0069] (3) dissolving the backbone polymer in 200 times the volume of methanol solution to obtain solution B;

[0070] (4) Solution A and Solution B were mixed by microfluidization technology, with the speed ratio of Solution A to Solution B being 1:1, and were repeatedly extruded by a 400 nm polycarbonate membrane extruder, and the organic solvent was removed by dialysis, and then freeze-dried to obtain a pharmaceutical composition containing telpotide.

[0071] Example 3: Pharmaceutical composition comprising telpotide

[0072] The present embodiment provides a pharmaceutical composition comprising tilpoitide, and the formula is as follows: polypeptide (tilpoitide 20 mg), backbone polymer (hyaluronic acid 20 mg, carboxymethyl chitosan 40 mg), enteric polymer (hydroxypropyl methylcellulose acetate succinate 30 mg, methacrylic acid-ethyl acrylate copolymer 10 mg, acrylic resin 20 mg), penetration enhancer (decanoic acid 10 mg, chenodeoxycholic acid 20 mg, 8-(2-hydroxybenzamido) caprylic acid 30 mg).

[0073] Preparation process:

[0074] (1) dissolving the polypeptide in a sodium hydroxide aqueous solution and adjusting the pH of the solution to 7.0, and dissolving the permeation enhancer and the enteric polymer in a sodium hydroxide aqueous solution and adjusting the pH of the solution to 7.0;

[0075] (2) Mixing the two solutions using a high-speed shearing machine for 20 minutes until uniformly mixed, to obtain solution A;

[0076] (3) dissolving the backbone polymer in 400 times the volume of acetone solution to obtain solution B;

[0077] (4) Solution A and Solution B were mixed by microfluidization technology, with the speed ratio of Solution A to Solution B being 3:2, and were repeatedly extruded by a 100 nm polycarbonate membrane extruder, and the organic solvent was removed by dialysis, and then freeze-dried to obtain a pharmaceutical composition containing telpotide.

[0078] Example 4-5: Selection of backbone polymer types in pharmaceutical compositions containing telpotide

[0079] Examples 4-5 provide two pharmaceutical compositions containing telpotide, which differ from Example 1 only in that 60 mg of the backbone polymer in Example 1 is replaced with hyaluronic acid and carboxymethyl chitosan of the same mass, respectively, and the remaining components and preparation process in the formula remain the same as in Example 1.

[0080] Example 6-8: Selection of enteric polymer type in pharmaceutical compositions containing telpotide

[0081] Examples 6-8 provide three pharmaceutical compositions containing telpotide, which differ from Example 1 only in that the 60 mg enteric polymer in Example 1 is replaced by 30 mg of hydroxypropyl methylcellulose acetate succinate + 30 mg of methacrylic acid-ethyl acrylate copolymer, 30 mg of hydroxypropyl methylcellulose acetate succinate + 30 mg of acrylic resin, and 30 mg of methacrylic acid-ethyl acrylate copolymer + 30 mg of acrylic resin, respectively, and the remaining components and preparation process in the formula remain consistent with Example 1.

[0082] Examples 9-11: Selection of penetration enhancers in pharmaceutical compositions containing telpotide

[0083] Examples 9-11 provide three pharmaceutical compositions containing telpotide, which differ from Example 1 in that the 60 mg permeation enhancer in Example 1 is replaced by 30 mg capric acid + 30 mg chenodeoxycholic acid, 30 mg capric acid + 30 mg 8-(2-hydroxybenzamido)octanoic acid, and 30 mg chenodeoxycholic acid + 30 mg 8-(2-hydroxybenzamido)octanoic acid, respectively, and the remaining components and preparation process in the formula remain consistent with Example 1.

[0084] Examples 12-14: Selection of the amount of polymer (backbone polymer + enteric polymer) in the pharmaceutical composition containing telpotide

[0085] Examples 12-14 provide three pharmaceutical compositions containing telportopide, which differ from Example 1 only in that the total amount of the backbone polymer and the enteric polymer is changed, the mass ratio of hyaluronic acid and carboxymethyl chitosan in the backbone polymer is maintained at 1:1, the mass ratio of hydroxypropyl methylcellulose acetate succinate, methacrylic acid-ethyl acrylate copolymer and acrylic resin in the enteric polymer is maintained at 1:1:1, and the remaining components and preparation process in the formula are consistent with Example 1. The specific formula is shown in Table 2:

[0086] Table 2

[0087]

[0088] Examples 15-17: Selection of the amount of backbone polymer in the pharmaceutical composition containing telpotide

[0089] Examples 15-17 provide three pharmaceutical compositions containing telportopide, which differ from Example 1 only in that the total amount of polymer (skeleton polymer + enteric polymer) is kept unchanged, and the mass percentage of the skeleton polymer in the polymer is adjusted. Among them, the mass ratio of hyaluronic acid and carboxymethyl chitosan in the skeleton polymer is maintained at 1:1, and the mass ratio of hydroxypropyl methylcellulose acetate succinate, methacrylic acid-ethyl acrylate copolymer and acrylic resin in the enteric polymer is maintained at 1:1:1, and the remaining components and preparation process in the formula are consistent with Example 1. The specific formula is shown in Table 3:

[0090] Table 3

[0091]

[0092]

[0093] Examples 18-21: Selection of the amount of penetration enhancer in a pharmaceutical composition containing telpotide

[0094] Examples 18-21 provide four pharmaceutical compositions containing telpotide, which differ from Example 1 in that the dosage of the penetration enhancer is adjusted, the mass ratio of decanoic acid, chenodeoxycholic acid and 8-(2-hydroxybenzamido)octanoic acid in the penetration enhancer is maintained at 1:1:1, and the remaining components and preparation process in the formula are consistent with those in Example 1. The specific formula is shown in Table 4:

[0095] Table 4

[0096]

[0097] Example 22: Pharmaceutical composition comprising semaglutide

[0098] The present embodiment provides a pharmaceutical composition comprising semaglutide, and the formula is as follows: polypeptide (semaglutide 20 mg), backbone polymer 60 mg (polylactic acid 30 mg, hydroxypropyl chitosan 30 mg), enteric polymer 60 mg (hydroxypropyl methylcellulose phthalate 20 mg, cellulose acetate phthalate 20 mg, acrylic resin 20 mg), penetration enhancer 60 mg (8-(2-hydroxybenzamido) caprylic acid 20 mg, ethylenediaminetetraacetic acid 20 mg, bile acid 20 mg).

[0099] The preparation process is consistent with that of Example 1.

[0100] Example 23: Pharmaceutical composition comprising semaglutide

[0101] The present embodiment provides a pharmaceutical composition comprising semaglutide, and the formula is as follows: polypeptide (semaglutide 20 mg), backbone polymer 60 mg (polylactic acid 40 mg, hydroxypropyl chitosan 20 mg), enteric polymer 60 mg (hydroxypropyl methylcellulose phthalate 15 mg, cellulose acetate phthalate 30 mg, acrylic resin 15 mg), penetration enhancer 60 mg (8-(2-hydroxybenzamido) caprylic acid 30 mg, ethylenediaminetetraacetic acid 15 mg, bile acid 15 mg).

[0102] The preparation process is consistent with that of Example 2.

[0103] Example 24: Pharmaceutical composition comprising semaglutide

[0104] The present embodiment provides a pharmaceutical composition comprising semaglutide, and the formula is as follows: polypeptide (semaglutide 20 mg), backbone polymer 60 mg (polylactic acid 20 mg, hydroxypropyl chitosan 40 mg), enteric polymer 60 mg (hydroxypropyl methylcellulose phthalate 30 mg, cellulose acetate phthalate 10 mg, acrylic resin 20 mg), penetration enhancer 60 mg (8-(2-hydroxybenzamido) caprylic acid 10 mg, ethylenediaminetetraacetic acid 20 mg, bile acid 30 mg).

[0105] The preparation process is consistent with that of Example 3.

[0106] Examples 25-26: Selection of backbone polymer types in pharmaceutical compositions containing semaglutide

[0107] Examples 25-26 provide two pharmaceutical compositions containing semaglutide, which differ from Example 22 only in that 60 mg of the backbone polymer in Example 22 is replaced with polylactic acid and hydroxypropyl chitosan of the same mass, respectively, and the remaining components and preparation process in the formula remain consistent with Example 22.

[0108] Examples 27-29: Selection of enteric polymer types in pharmaceutical compositions containing semaglutide

[0109] Examples 27-29 provide three pharmaceutical compositions containing semaglutide, which differ from Example 22 only in that: the 60 mg enteric polymer in Example 22 is replaced by 30 mg hydroxypropyl methylcellulose phthalate + 30 mg cellulose acetate phthalate, 30 mg hydroxypropyl methylcellulose phthalate + 30 mg acrylic resin, and 30 mg cellulose acetate phthalate + 30 mg acrylic resin, respectively, and the remaining components and preparation process in the formula remain consistent with Example 22.

[0110] Examples 30-32: Selection of penetration enhancers in pharmaceutical compositions containing semaglutide

[0111] Examples 30-32 provide three pharmaceutical compositions containing semaglutide, which differ from Example 22 only in that the 60 mg penetration enhancer in Example 22 is replaced by 8-(2-hydroxybenzamido)octanoic acid 30 mg+ethylenediaminetetraacetic acid 30 mg, 8-(2-hydroxybenzamido)octanoic acid 30 mg+cholic acid 30 mg, and ethylenediaminetetraacetic acid 30 mg+cholic acid 30 mg, respectively, and the remaining components and preparation process in the formula remain consistent with Example 22.

[0112] Example 33: Pharmaceutical composition comprising liraglutide

[0113] The present embodiment provides a pharmaceutical composition comprising liraglutide, and the formula is as follows: polypeptide (20 mg of liraglutide), 60 mg of backbone polymer (30 mg of alginate, 30 mg of sodium hyaluronate), 60 mg of enteric polymer (20 mg of methacrylic acid-ethyl acrylate copolymer, 20 mg of cellulose acetate phthalate, 20 mg of hydroxypropyl methylcellulose phthalate), and 60 mg of penetration enhancer (20 mg of caprylic acid, 20 mg of capric acid, and 20 mg of citric acid).

[0114] The preparation process is consistent with that of Example 1.

[0115] Example 34: Pharmaceutical composition comprising liraglutide

[0116] The present embodiment provides a pharmaceutical composition comprising liraglutide, and the formula is as follows: polypeptide (20 mg of liraglutide), 60 mg of backbone polymer (40 mg of alginate, 20 mg of sodium hyaluronate), 60 mg of enteric polymer (15 mg of methacrylic acid-ethyl acrylate copolymer, 30 mg of cellulose acetate phthalate, 15 mg of hydroxypropyl methylcellulose phthalate), and 60 mg of penetration enhancer (40 mg of caprylic acid, 10 mg of capric acid, and 10 mg of citric acid).

[0117] The preparation process is consistent with that of Example 2.

[0118] Example 35: Pharmaceutical composition comprising liraglutide

[0119] The present embodiment provides a pharmaceutical composition comprising liraglutide, and the formula is as follows: polypeptide (20 mg of liraglutide), 60 mg of backbone polymer (20 mg of alginate, 40 mg of sodium hyaluronate), 60 mg of enteric polymer (30 mg of methacrylic acid-ethyl acrylate copolymer, 10 mg of cellulose acetate phthalate, 20 mg of hydroxypropyl methylcellulose phthalate), and 60 mg of penetration enhancer (10 mg of caprylic acid, 20 mg of capric acid, and 30 mg of citric acid).

[0120] The preparation process is consistent with that of Example 3.

[0121] Examples 36-37: Selection of backbone polymer types in pharmaceutical compositions containing liraglutide

[0122] Examples 36-37 provide two pharmaceutical compositions containing liraglutide, which differ from Example 33 only in that 60 mg of the backbone polymer in Example 33 is replaced with alginate and sodium hyaluronate of the same mass, respectively, and the remaining components and preparation process in the formula remain consistent with Example 33.

[0123] Examples 38-40: Selection of enteric polymer types in pharmaceutical compositions containing liraglutide

[0124] Examples 38-40 provide three pharmaceutical compositions containing liraglutide, which differ from Example 33 only in that: the 60 mg enteric polymer in Example 33 is replaced by 30 mg methacrylic acid-ethyl acrylate copolymer + 30 mg cellulose acetate phthalate, 30 mg methacrylic acid-ethyl acrylate copolymer + 30 mg hydroxypropyl methylcellulose phthalate, and 30 mg cellulose acetate phthalate + 30 mg hydroxypropyl methylcellulose phthalate, respectively, and the remaining components and preparation process in the formula are consistent with Example 33.

[0125] Examples 41-43: Selection of penetration enhancer types in pharmaceutical compositions containing liraglutide

[0126] Examples 41-43 provide three pharmaceutical compositions containing liraglutide, which differ from Example 33 in that the 60 mg penetration enhancer in Example 33 is replaced by 30 mg caprylic acid + 30 mg capric acid, 30 mg caprylic acid + 30 mg citric acid, and 30 mg capric acid + 30 mg citric acid, respectively, and the remaining components and preparation process in the formula remain consistent with Example 33.

[0127] Comparative Examples 1-5

[0128] Comparative Examples 1-5 provide five pharmaceutical compositions containing telpotide, which differ from Example 1 in that, respectively, enteric polymer + skeleton polymer + permeation enhancer, enteric polymer + skeleton polymer, skeleton polymer, enteric polymer, permeation enhancer are not added to the compositions, and the formulas of the comparative examples are shown in Table 5:

[0129] Table 5

[0130] Telportide Backbone polymer Enteric polymers Penetration enhancers Example 1 20mg 60mg 60mg 60mg Comparative Example 1 20mg / / / Comparative Example 2 20mg / / 60mg Comparative Example 3 20mg / 120mg 60mg Comparative Example 4 20mg 120mg / 60mg Comparative Example 5 20mg 60mg 60mg /

[0131] Preparation process:

[0132] In Comparative Example 1, 20 mg of the polypeptide active substance was directly used for subsequent research and testing;

[0133] The difference between the preparation process of Comparative Example 2 and Example 1 is that, in step (1), no enteric polymer is added and step (3) is not performed, and the solution A is freeze-dried to obtain a pharmaceutical composition containing telpotide;

[0134] The difference between the preparation process of Comparative Example 3 and Example 1 is that step (3) is not performed, and the solution A is freeze-dried to obtain a pharmaceutical composition containing telpotide;

[0135] The difference between the preparation processes of Comparative Examples 4 and 5 and Example 1 is that, respectively, no enteric polymer and no penetration enhancer are added in step (1).

[0136] Comparative Examples 6-10

[0137] Comparative Examples 6-10 provide five pharmaceutical compositions containing semaglutide, which differ from Example 22 only in that enteric polymer + skeleton polymer + permeation enhancer, enteric polymer + skeleton polymer, skeleton polymer, enteric polymer, permeation enhancer are not added to the compositions, respectively. The formulations of each comparative example are shown in Table 6:

[0138] Table 6

[0139] Semaglutide Backbone polymer Enteric polymers Penetration enhancers Embodiment 22 20mg 60mg 60mg 60mg Comparative Example 6 20mg / / / Comparative Example 7 20mg / / 60mg Comparative Example 8 20mg / 120mg 60mg Comparative Example 9 20mg 120mg / 60mg Comparative Example 10 20mg 60mg 60mg /

[0140] The preparation process of Comparative Examples 6-10 refers to Comparative Examples 1-5.

[0141] Comparative Examples 11-15

[0142] Comparative Examples 11-15 provide five pharmaceutical compositions containing liraglutide, which are different from Example 33 only in that enteric polymer + skeleton polymer + permeation enhancer, enteric polymer + skeleton polymer, skeleton polymer, enteric polymer, permeation enhancer are not added to the compositions, respectively. The formulas of each comparative example are shown in Table 7:

[0143] Table 7

[0144] Liraglutide Backbone polymer Enteric polymers Penetration enhancers Embodiment 33 20mg 60mg 60mg 60mg Comparative Example 11 20mg / / / Comparative Example 12 20mg / / 60mg Comparative Example 13 20mg / 120mg 60mg Comparative Example 14 20mg 120mg / 60mg Comparative Example 15 20mg 60mg 60mg /

[0145] The preparation process of Comparative Examples 11-15 refers to Comparative Examples 1-5.

[0146] Application Example 1

[0147] Application Example 1 provides an oral tablet containing telpotide.

[0148] The oral tablet has a formula of: the pharmaceutical composition prepared in Example 1 (200 mg), excipients (196 mg lactose, 4 mg magnesium stearate), with a total mass of 400 mg.

[0149] Preparation method: The pharmaceutical composition and excipients are mixed evenly, and pressed into 150 mg round tablets. The hardness of the tablets is 80N and the brittleness is 0.4%.

[0150] Application Example 2

[0151] Application Example 2 provides an oral granule containing telpotide.

[0152] The oral granules have a formula of: the pharmaceutical composition prepared in Example 1 (200 mg), excipients (196 mg lactose, 4 mg magnesium stearate), with a total mass of 400 mg. The pharmaceutical composition and excipients are granulated and mixed to obtain oral granules.

[0153] Comparative Application Example 1-Comparative Application Example 10

[0154] Comparative Application Examples 1 to 5 each provide an oral tablet containing telpotide, and Comparative Application Examples 6 to 10 each provide an oral granule containing telpotide. The formula of each comparative application example is shown in Table 8:

[0155] Table 8

[0156]

[0157] The preparation process of the oral tablets of Comparative Application Examples 1-5 is consistent with that of Application Example 1; the preparation process of the oral granules of Comparative Application Examples 6-10 is consistent with that of Application Example 2.

[0158] Test Example 1

[0159] In vitro permeability coefficient Papp test:

[0160] After culture, the Caco-2 cell monolayer differentiated into the apical side of the villus and the basal side of the basal side, exhibited cell polarity, and had a clearly defined apical brush border and tight intercellular junctions as well as active transporters in the small intestine, morphologically similar to small intestinal absorptive cells.

[0161] After 21 days of culture, Caco-2 cells were built into an in vitro Transwell transport chamber. After the integrity of the Caco-2 cell monolayer was confirmed to be qualified by transmembrane resistance, alkaline phosphatase activity, and fluorescent yellow permeability test results, a transport test was performed.

[0162] The Caco-2 cell monolayer model was washed three times with HBSS buffer (37°C), HBSS buffer was added, incubated for 20 minutes, and the supernatant was discarded and the bidirectional transport test was immediately started. To investigate the transport from the apical side (AP side) to the basal side (BL side), 0.5 mL of PBS solution containing 0.05 mg / mL of the drug composition was added to the AP side as the test solution, and 1.5 mL of blank PBS solution was added to the BL side. The mixture was placed in a constant temperature shaker (37°C, 40 r / min) and incubated. At 60 minutes, 200 μL of solution was drawn from the BL side as the test sample, and 200 μL of blank PBS solution was added at the same time. When investigating the transport from the BL side to the AP side, 1.5 mL of PBS solution containing the drug composition was added to the BL side as the test solution, and 0.5 mL of blank PBS solution was added to the AP side. The mixture was placed in a constant temperature shaker (37°C, 40 r / min) and incubated. At 60 minutes, 200 μL of solution was drawn from the AP side as the test sample, and 200 μL of blank PBS solution was added at the same time. The sample was taken for content determination and the apparent permeability coefficient Papp was calculated:

[0163] Papp = (ΔQ / Δt) / (A×C0);

[0164] Where ΔQ is the transport amount within Δt, A is the surface membrane area of ​​the 12-well plate, and C0 is the initial mass concentration in the supply chamber.

[0165] The calculation formula of transfer volume (Q) is:

[0166] AP→BL: QB=1.5×Cr;

[0167] BL→AP: QA=0.5×Cr;

[0168] Wherein, 0.5 is the volume of AP chamber, unit is mL; 1.5 is the volume of BL chamber, unit is mL; Cr is the drug concentration at 60 min.

[0169] The permeability of the polypeptides in different pharmaceutical compositions is shown in Table 9:

[0170] Table 9

[0171]

[0172]

[0173] It can be seen from the data of Example 1 and Comparative Example 1 that the permeability of telpotide alone is extremely poor, which further indicates that after telpotide enters the gastrointestinal tract, it cannot penetrate the intestinal surface cells and has a very low bioavailability; it can be seen from the data of Example 1 and Comparative Example 5 that the addition of a permeation enhancer to the pharmaceutical composition greatly improves the permeability of telpotide.

[0174] It can be seen from the data in Examples 1 and 9 to 11 that capric acid, chenodeoxycholic acid and 8-(2-hydroxybenzamido)octanoic acid cooperate with each other to play a synergistic role in improving the permeability of tepote in the composition; it can be seen from the data in Examples 1 and 18 to 21 that the permeation enhancer has a more obvious improvement effect on the permeability of tepote when the amount of the permeation enhancer is controlled at 15% to 30%.

[0175] It can be seen from the data of Example 22 and Comparative Example 6 that the permeation effect of semaglutide alone is very poor; it can be seen from the data of Example 22 and Comparative Example 10 that the permeation ability of semaglutide is greatly increased by adding a permeation enhancer to the pharmaceutical composition; it can be seen from the data in Example 22 and Examples 30 to 32 that 8-(2-hydroxybenzamido)octanoic acid, ethylenediaminetetraacetic acid and bile acid cooperate with each other and play a synergistic role in improving the permeation effect of semaglutide in the composition.

[0176] It can be seen from the data of Example 33 and Comparative Example 11 that the permeation effect of liraglutide alone is very poor; it can be seen from the data of Example 33 and Comparative Example 15 that the permeation ability of liraglutide is greatly increased by adding a permeation enhancer to the pharmaceutical composition; it can be seen from the data in Example 33 and Examples 41 to 43 that caprylic acid, capric acid and citric acid cooperate with each other and play a synergistic role in improving the permeation effect of liraglutide in the composition.

[0177] Test Example 2

[0178] Dissolution test:

[0179] According to the sixth method (flow cell method) of 0931 of the fourth general rules of the Chinese Pharmacopoeia in 2020, the sample is first dissolved in a pH 1.0 hydrochloric acid solution for 2 hours, and then dissolved in a pH 6.0 phosphate buffer for 0.25 hours and / or 0.5 hours. The dissolution medium, pH 1.0 hydrochloric acid solution, is a 0.1 mol / L hydrochloric acid aqueous solution, and the dissolution medium, pH 6.0 phosphate buffer, is a 0.05 mol / L phosphate buffer. Sampling is performed at specified time points (for example, pH 1.0 is 2 hours, pH 6.0 is 0.25 hours and / or 0.5 hours), and the HPLC method is used to detect the polypeptide content and the total content of multiple penetration enhancers, and the percentage content is calculated according to the labeled amount of the polypeptide and penetration enhancer in the composition / tablet / granule, which is its dissolution amount.

[0180] (1) The dissolution amounts of polypeptides and penetration enhancers in different compositions are shown in Table 10:

[0181] Table 10

[0182]

[0183]

[0184] From the data of Example 1 / Example 22 / Example 33 and Comparative Examples 1-4 / Comparative Examples 6-9 / Comparative Examples 11-14, it can be seen that enteric polymers and backbone polymers can significantly reduce the dissolution amount of polypeptides and permeation enhancers in gastric juice and increase the dissolution amount in intestinal juice, indicating that the pharmaceutical composition containing bioactive substances prepared by the present invention can remain stable in artificial gastric juice and can be rapidly dissolved in intestinal juice.

[0185] It can be seen from the data of Example 1 and Examples 4 to 5 that in the pharmaceutical composition containing telpotide, the backbone polymers hyaluronic acid and carboxymethyl chitosan have a significant synergistic effect in reducing the dissolution amount in gastric juice and increasing the dissolution amount in intestinal juice; it can be seen from the data of Example 1 and Examples 6 to 8 that in the pharmaceutical composition containing telpotide, the enteric polymers hydroxypropyl methylcellulose acetate succinate, methacrylic acid-ethyl acrylate copolymer and acrylic resin have a significant synergistic effect in reducing the dissolution amount in gastric juice and increasing the dissolution amount in intestinal juice.

[0186] From the data of Example 1 and Examples 12 to 14, it can be seen that by controlling the mass percentage of the polymer in the composition to 55% to 70%, telpoitide can be further maintained stable in gastric juice and rapidly released in intestinal juice; from the data of Example 1 and Examples 15 to 17, it can be seen that by controlling the mass percentage of the backbone polymer in the polymer to 45% to 55%, telpoitide can be further maintained stable in gastric juice and rapidly released in intestinal juice.

[0187] It can be seen from the data of Example 22 and Examples 25-26 that in the pharmaceutical composition containing semaglutide, the backbone polymers polylactic acid and hydroxypropyl chitosan have a significant synergistic effect in reducing the dissolution amount in gastric juice and increasing the dissolution amount in intestinal juice; it can be seen from the data of Example 22 and Examples 27-29 that in the pharmaceutical composition containing semaglutide, the enteric polymers hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate and acrylic resin have a significant synergistic effect in reducing the dissolution amount in gastric juice and increasing the dissolution amount in intestinal juice.

[0188] It can be seen from the data of Example 33 and Examples 36 to 37 that in the pharmaceutical composition containing liraglutide, the backbone polymers alginate and sodium hyaluronate have a significant synergistic effect in reducing the dissolution amount in gastric juice and increasing the dissolution amount in intestinal juice; it can be seen from the data of Example 33 and Examples 38 to 40 that in the pharmaceutical composition containing liraglutide, the enteric polymers methacrylic acid-ethyl acrylate copolymer, cellulose acetate phthalate and hydroxypropyl methylcellulose phthalate have a significant synergistic effect in reducing the dissolution amount in gastric juice and increasing the dissolution amount in intestinal juice.

[0189] (2) The dissolution amounts of polypeptides and penetration enhancers in different tablets / granules are shown in Table 11:

[0190] Table 11

[0191]

[0192] It can be seen from the data in Application Examples 1 / 2 that the tablets / granules further prepared from the pharmaceutical composition containing telpotide of the present invention have very low dissolution amounts of telpotide and permeation enhancer in artificial gastric juice, while the dissolution amounts in artificial intestinal juice are significantly increased, indicating that the tablets / granules containing telpotide have good stability in gastric juice and can be quickly dissolved in artificial intestinal juice.

[0193] It can be seen from the data in Application Example 1 / 2 and Comparative Application Example 1 / 6 that when the composition is composed only of telpotide, telpotide in the further prepared tablets / granules has been dissolved in the artificial gastric juice, and telpotide is unstable and has been degraded.

[0194] It can be seen from the data in Application Examples 1 / 2 and Comparative Application Examples 2 to 4 / 7 to 9 that when the composition does not contain a backbone polymer, the dissolution amount of telpotide in the further prepared tablets / granules is greatly reduced; when the composition does not contain an enteric polymer, in the further prepared tablets / granules, telpotide is completely dissolved in artificial gastric juice and has been degraded, and has poor stability.

[0195] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0196] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0197] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A pharmaceutical composition comprising a biologically active substance, characterized in that: The pharmaceutical composition comprises a bioactive substance, a polymer and a penetration enhancer; The bioactive substance is selected from telpotide, semaglutide or liraglutide; The polymers include backbone polymers and enteric polymers.

2. The pharmaceutical composition according to claim 1, characterized in that Calculated by mass percentage, the pharmaceutical composition comprises 5-30% of biologically active substances, 55-70% of polymers and 15-30% of penetration enhancers; Preferably, the mass percentage of the backbone polymer in the polymer is 45-55%.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that The bioactive substance is selected from telportin; the backbone polymer includes hyaluronic acid and carboxymethyl chitosan; the enteric polymer includes hydroxypropyl methylcellulose acetate succinate, methacrylic acid-ethyl acrylate copolymer and acrylic resin; the penetration enhancer includes capric acid, chenodeoxycholic acid and 8-(2-hydroxybenzamido)octanoic acid; Preferably, the mass ratio of hyaluronic acid to carboxymethyl chitosan is 1:(0.1-1); Preferably, the mass ratio of the hydroxypropyl methylcellulose acetate succinate, the methacrylic acid-ethyl acrylate copolymer and the acrylic resin is 1:(0.1-1):(0.1-1); Preferably, the mass ratio of decanoic acid, chenodeoxycholic acid and 8-(2-hydroxybenzamido)octanoic acid is 1:(0.1-1):(0.1-1).

4. The pharmaceutical composition according to claim 1 or 2, characterized in that The bioactive substance is selected from semaglutide; the skeleton polymer includes polylactic acid and hydroxypropyl chitosan; the enteric polymer includes hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate and acrylic resin; the penetration enhancer includes 8-(2-hydroxybenzamido)octanoic acid, ethylenediaminetetraacetic acid and bile acid; Preferably, the mass ratio of the polylactic acid to hydroxypropyl chitosan is 1:(0.1-1); Preferably, the mass ratio of hydroxypropyl methylcellulose phthalate, cellulose acetate phthalate and acrylic resin is 1:(0.1-1):(0.1-1); Preferably, the mass ratio of 8-(2-hydroxybenzamido)octanoic acid, ethylenediaminetetraacetic acid and cholic acid is 1:(0.1-1):(0.1-1).

5. The pharmaceutical composition according to claim 1 or 2, characterized in that The bioactive substance is selected from liraglutide; the backbone polymer includes alginate and sodium hyaluronate; the enteric polymer includes methacrylic acid-ethyl acrylate copolymer, cellulose acetate phthalate and hydroxypropyl methylcellulose phthalate; the penetration enhancer includes caprylic acid, capric acid and citric acid; Preferably, the mass ratio of alginate to sodium hyaluronate is 1:(0.1-1); Preferably, the mass ratio of the methacrylic acid-ethyl acrylate copolymer, cellulose acetate phthalate and hydroxypropyl methylcellulose phthalate is 1:(0.1-1):(0.1-1); Preferably, the mass ratio of caprylic acid, capric acid and citric acid is 1:(0.1-1):(0.1-1).

6. A method for preparing a pharmaceutical composition comprising a biologically active substance as claimed in any one of claims 1 to 5, characterized in that: The method comprises the following steps: (1) dissolving the bioactive substance, the enteric polymer and the permeation enhancer in solvent I to obtain solution A; dissolving the backbone polymer in solvent III to obtain solution B; (2) Solution A and solution B are mixed and extruded to obtain a pharmaceutical composition solution, which is then dried to obtain a pharmaceutical composition containing a bioactive substance.

7. The preparation method according to claim 6, characterized in that: The solvent I is an alkaline solution, preferably an aqueous sodium hydroxide solution; Preferably, the solvent II comprises any one of methanol, ethanol, isopropanol or acetone, or a combination of at least two thereof; Preferably, the mixing in step (2) includes mixing using microfluidization technology.

8. An oral pharmaceutical preparation containing a biologically active substance, characterized in that: The pharmaceutical preparation comprises the pharmaceutical composition containing the bioactive substance according to any one of claims 1 to 5, and a pharmaceutically acceptable excipient.

9. The oral pharmaceutical preparation according to claim 8, characterized in that In the oral pharmaceutical preparation, the mass percentage of the pharmaceutical composition is 5 to 70%; Preferably, the excipient comprises any one of lactose, magnesium stearate, mannitol or microcrystalline cellulose, or a combination of at least two thereof.

10. The oral pharmaceutical preparation according to claim 8 or 9, characterized in that The dosage form of the pharmaceutical preparation is a solid or semisolid preparation; Preferably, the solid preparation includes tablets, granules or capsules; Preferably, the semisolid preparation comprises a suspension or an emulsion.

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