Supramolecular nanocomposite and use

Abstract

A supramolecular nanocomposite, characterized in that the supramolecular nanocomposite comprises an active substance, and a flavonoid glycoside and a terpenoid glycoside serving as carriers. The supramolecular nanocomposite can significantly improve the absorption of poorly absorbable substances in the colon / rectum at the site.

Description

Supramolecular nanocomplexes and uses thereof TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to a supramolecular nanocomplex for improving the absorption of active substances in the rectum and / or colon and uses thereof. BACKGROUND

[0002] According to Pharmaceutics.2021 Feb 17;13(2):272, the gastrointestinal tract can be divided into several anatomical compartments from the esophagus, stomach, small intestine (including the duodenum, jejunum and ileum) and large intestine (colon and rectum) from the anatomical point of view. These compartments have significant differences in length, absorption area, drug residence and transport time, liquid composition, pH value, mucus layer thickness and type, variable cell type, transport protein expression, cytochrome isozyme P450 expression, permeability, tight junction resistance, resident microflora and abundance, etc. Changes in any one or more of these factors will directly affect the dissolution, transport and absorption of drugs in the gastrointestinal tract after oral administration. According to Nat Rev Gastroenterol Hepatol.2022 Apr;19(4):219-238, the transport time of drugs in the gastrointestinal tract is one of the limiting factors for the absorption of orally administered drugs. The transport time of drugs in the small intestine after oral administration is 1-6h, and the transport time in the colon is usually up to 1-3 days, which is much higher than that in other anatomical compartments. The transport time of drugs in the colon, the liquid composition in the colon and the microflora are often referred to as the colon variability factor. The colon variability factor is both an opportunity and a challenge for the development of new oral sustained-release or long-acting formulations.

[0003] (1) Low colon absorption drugs

[0004] Connor A, King G, Jones K. Evaluation of human regional bioavailability to assess whether modified release development is feasible, https: / / cdn.technologynetworks.com / TN / Resources / PDF / 510.pdf 2014 reviewed the results of 37 human regional drug absorption studies. About 65% of new chemical entities (NCEs) drugs and 53% of marketed drugs had a relative bioavailability (Fcolon = AUCIR / AUCcolon * 100%) of less than 60% after colon administration; about 59% of new chemical entities (NCEs) drugs and 21% of marketed drugs had a relative bioavailability (Fcolon = AUCIR / AUCcolon * 100%) of less than 30% after colon administration.

[0005] For example, the apparent half-life of Apixaban is 12 hours, in theory, it can be developed into a once-a-day preparation. But according to Apixaban tablet FDA review report CLINICAL PHARMACOLOGY AND BIOPHARMACEUTICS REVIEW(S) P98-100: Study CV185007 (Regional GI absorption), the pharmacokinetic results of different parts of the digestive tract are as follows:

[0006] Table 1. Pharmacokinetic results of different parts of the digestive tract of Apixaban Note: Cmax, AUC are geometric means, CV% in parentheses; T 1 / 2 Mean, SD in parentheses; Tmax is the median value, the range in parentheses.

[0007] According to the above research results, the plasma exposure of Apixaban (AUC 0-t ) of the solution of Apixaban administered in the ascending colon is about 17.25% of the oral exposure (Fcolon = 73.5 / 426.1 x 100% = 17.25%), indicating that the absorption in the colon is significantly lower than that in the upper digestive tract; compared with the solution, the plasma exposure of Apixaban (AUC 0-t ) of the crushed Apixaban tablet administered in the ascending colon is reduced by 67.3%, indicating that solubility is also a key factor affecting the absorption of Apixaban in the colon. The multi-factor effect led to the failure of the development of the sustained-release preparation of Apixaban by Pfizer / Pfizer / Bristol-Myers Squibb, resulting in the final marketed Apixaban tablet Eliquis / Eliquis Apixaban tablet, which is administered twice a day, see patent CN102802608A. Therefore, low straight colon absorption drugs, even through the existing technology such as gastric floating, gastric retention, etc. to prolong the release and transport time of the drug in the upper digestive tract, it is difficult to provide and maintain effective systemic exposure and exposure concentration within the drug interval, resulting in treatment or development failure.

[0008] (2) BCS III (high solubility and low permeability), IV drug (low solubility and low permeability), the drug has lower intestinal permeability and slower absorption, and if it is superimposed with lower straight / colon absorption, it will lead to a significant reduction in the bioavailability of the drug.

[0009] (3) Metabolic enzymes, transport proteins in the colon have greater differences compared with the small intestine. According to Journal of Controlled Release 353 (2023) 1107-1126, the activities of P-gp efflux protein, BCRP, MRP2, P450 cytochrome isozyme, PEPT1 in the colon are significantly lower than those in the duodenum and small intestine, while the activities of MRP3, MCT1, OATPB, OCTN2, OCT1 are significantly higher than those in the duodenum and small intestine, which provides a better solution in terms of compliance, safety and effectiveness for drugs that must be administered in large doses or invasively to achieve clinical treatment due to significant first-pass metabolism of P450 enzymes or P-gp efflux.

[0010] (4) Generally, oral drug delivery systems treat drugs that are absorbed by the small intestinal mucosa after entering the intestinal tract, enter the small intestinal capillaries, and converge in the superior mesenteric vein before entering the liver portal vein, and then enter the liver. After metabolism by the liver, it enters the main circulation and circulates to the body organs and tissues through the blood. The absorption pathways of rectal drug delivery mainly include three:

[0011] ① Superior rectal vein → portal vein → liver → main circulation;

[0012] ② Inferior rectal vein + anal vein → internal iliac vein (bypassing the liver) → inferior vena cava → main circulation;

[0013] ③ Rectal lymphatic system.

[0014] Therefore, drugs that are severely affected by first-pass metabolism / efflux or have target organs in the lymphatic system can bypass first-pass metabolism / efflux by rectal / colonic drug delivery, thereby avoiding invasive drug delivery.

[0015] The above factors result in significant differences in the in vivo pharmacokinetic data of drugs absorbed by the rectum / colon and drugs absorbed by the small intestine. If the differentiated pharmacokinetic characteristics are desired for disease treatment, low rectal / colonic drug absorption will make it difficult to achieve this goal. However, the above (1), (2), (3), and (4) are often superimposed and difficult to separate.

[0016] For example, according to J Drug Assess. 2014 Aug 12; 3(1): 43-50, the pharmacokinetic results of different drug delivery sites of ticagrelor are shown in Table 2:

[0017] Table 2. Pharmacokinetic results of different drug delivery sites of ticagrelor Note: CV% in parentheses, or range

[0018] The plasma exposure (AUC0-24) of ticagrelor suspension in the proximal small intestine, distal small intestine, and ascending colon was 0.67, 0.59, and 0.56 pg / mL, respectively, while the plasma exposure (AUC0-24) of the prototype drug and metabolite AR-C124910XX was 0.67, 0.59, and 0.56 pg / mL, respectively.0-t The ratios of the original drug ticagrelor to its metabolite AR-C124910XX were calculated according to the table above for administration to different sites. The results are shown in Table 3 below.

[0019] Table 3. Differences between the parent drug and metabolite AR-C124910XX of ticagrelor administered at different sites

[0020] Based on the above results, it can be seen that the metabolite / original AUC varies depending on the administration site. 0-t The ratios showed significant differences, with the AUC for colonic administration being significantly different. 0-t The lowest ratio, consistent with intravenous administration, indicates that colonic administration bypasses first-pass metabolism. However, if the drug itself has low absorption in the colon / rectum, and there is no corresponding technology to address this, it limits the achievement of this goal. For example, ticagrelor, with an Fc of 1119 / 3781*100% = 29.6%, restricts the development of rectal / colonic formulations.

[0021] To address the absorption challenges of rectal / colonic drug delivery systems, existing technologies such as gastric retention extend the drug's residence time in the stomach, thereby prolonging absorption in the upper digestive tract. Several related products have been successfully marketed, including Glumetza, Janumet XR, Baclofen GRS, and Cifran. However, the gastric retention time of gastric retention technology varies greatly and is easily affected by changes in external conditions, leading to gastric retention failure and premature passage of the drug into the intestines. Subsequent attempts have also focused on controlling the time it takes for a solid dosage form to pass through the pylorus, but this results in significant individual differences in retention time due to variations in pyloric opening size and gastric motility, and also carries the risk of pyloric obstruction.

[0022] Currently, the goal of achieving differentiated drug metabolism can only be partially achieved by changing the route of administration, such as injection or transdermal administration. However, these methods have significant limitations, with some routes of administration resulting in poor patient compliance and a high risk of infection.

[0023] Therefore, there is a great clinical need for a rectal / colonic hyperabsorption drug delivery technology to enable the absorption of low-absorption drugs in the rectum / colon at that site, thereby improving patient compliance, efficacy, and safety.

[0024] Summary of the Invention

[0025] Technical solution of the present invention:

[0026] Supermolecular nanocomplexes for improving the absorption of active substances in the rectum and / or colon, characterized in that the supermolecular nanocomplexes comprise an active substance and as carrier flavonoid glycosides and terpene glycosides, preferably the active substance is a drug, a diagnostic agent, a nutritional supplement, wherein the bioavailability (area under the plasma or serum or whole blood concentration-time curve AUC) after oral administration of an oral formulation of the active substance compared to the bioavailability (area under the plasma or serum or whole blood concentration-time curve AUC) after colonic and / or rectal administration of the oral formulation is F 结肠 or F 直肠 ≤ 60% (preferably ≤ 40%), wherein F 结肠 = AUC 结肠 / AUC 口服 x 100%; F 直肠 = AUC 直肠 / AUC 口服 x 100%.

[0027] Colon, rectum low-absorbing substances (such as drugs, diagnostic agents, nutritional supplements):

[0028] In the case of drugs, colon and / or rectum (or indicated as "rectum / colon" or "colon / rectum") low-absorbing drugs according to the present application mean that the bioavailability (area under the plasma or serum or whole blood concentration-time curve AUC) after colonic or rectal administration of an oral formulation compared to the bioavailability (area under the plasma or serum or whole blood concentration-time curve AUC) after oral administration of the oral formulation is F 结肠 or F 直肠 ≤ 60% (preferably ≤ 40%). F 结肠 = AUC 结肠 / AUC 口服 x 100%; F 直肠 = AUC 直肠 / AUC 口服 x 100%.

[0029] The colon / rectum low absorbed drug is preferably one or more of apixaban, rivaroxaban, edoxaban, ticagrelor, dabigatran, carvedilol, zafirlukast, cyclosporine, nitrendipine, gabapentin, pregabalin, cannabidiol, acitretin, tacrolimus, sirolimus, metformin, imeglimin, afatinib, bosutinib, colchicine, everolimus, ledipasvir, naloxegol, budesonide, bromocriptine, haloperidol, doxorubicin, felodipine, isradipine, fluvastatin, donetumab, ketamine, morphine, pyridostigmine, naloxone, naltrexone, thioridazine, selegiline, tacrine, terbutaline, lidocaine, pazopanib, prucalopride, rifaximin, ranolazine, furosemide, ritonavir, baclofen, captopril, benazepril, sumatriptan, cimetidine, ranitidine, fexofenadine, atenolol, ciprofloxacin, silodosin, prostat, acrivastine, ibutilide, simvastatin, lovastatin, pravastatin, atorvastatin, rosuvastatin, halofuginol, cinnarizine, griseofulvin, paclitaxel, docetaxel, topotecan, nimodipine, nisoldipine, nicardipine, fenofibrate, mycophenolate mofetil, mycophenolic acid, fenofibrate ester, chlorphenesin, probucol, tamsulosin, alfuzosin, finasteride, alendronate, clodronate, etidronate, pamidronate, risedronate, zoledronate.

[0030] Flavonoid glycoside:

[0031] The flavonoid glycoside in the present application is a flavonoid glycoside of the structure of Formula I.

[0032] R1 in the flavonoid glycoside of Formula I is a sugar residue, which is one of monosaccharide, disaccharide, oligosaccharide, polysaccharide, wherein R2, R3, R4 are one of H, OH, OR5, and R5 is one of C1-C10 alkane or one of monosaccharide, disaccharide, oligosaccharide, polysaccharide.

[0033] The flavonoid glycoside of Formula I is preferably one or more of naringin dihydrochalcone, neohesperidin dihydrochalcone, trilobatin, phloridzin, aspartame, 1-(3-beta-D-glucopyranosyl-2,4,6-trihydroxyphenyl)-3-(4-hydroxyphenyl)-1-propanone. The flavonoid glycoside of Formula I is more preferably naringin dihydrochalcone, neohesperidin dihydrochalcone.

[0034] Terpenoid glycoside

[0035] The terpenoid glycoside in the present application is one or more of a terpenoid glycoside of Formula II, a terpenoid glycoside of Formula III, a terpenoid glycoside of Formula IV.

[0036] Terpenoid glycoside of Formula II:

[0037] The terpenoid glycoside of Formula II is a terpenoid glycoside of the following structure.

[0038] R6is a hydroxyl group, a sugar residue, R7is H, a methyl group, an ethyl group, an isopropyl group, an alkali metal ion, or a sugar residue, and the sugar residue is independently one of a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide. The terpene glycoside of Formula II can be used singly or as a mixture.

[0039] The terpene glycoside of Formula II is preferably one or more of stevioside, rebaudioside A, rebaudioside B and salts thereof, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside M, rebaudioside N, rebaudioside O, dulcoside A, dulcoside B, rubusoside, steviol, steviolbioside and salts thereof. More preferably, it is one or more of rebaudioside A, stevioside, rebaudioside B and salts thereof, rebaudioside C.

[0040] The terpene glycoside of Formula III is a terpene glycoside of the following structure.

[0041] The terpene glycoside of Formula III is a terpene glycoside of the following structure.

[0042] R8and R9are H or a sugar residue, and are not H at the same time. The sugar residue is independently one of a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide. The terpene glycoside of Formula III can be used singly or as a mixture.

[0043] The terpene glycoside of Formula III is preferably one or more of mogroside V, mogroside IV, mogroside IIA1, mogroside IIA2, mogroside VIA, mogroside VIB, mogroside III, mogroside I, siamenoside I.

[0044] The terpene glycoside of Formula IV is a terpene glycoside of the following structure.

[0045] The terpene glycoside of Formula IV is a terpene glycoside of the following structure.

[0046] R10is H, an alkali metal ion, or a sugar residue, R11is H, an alkali metal ion, an organic base, a uronic acid, or a sugar residue. R12is H, a glucuronic acid and an alkali metal salt, an ammonium salt, or a sugar residue thereof. The sugar residue is independently one of a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide. The terpene glycoside of Formula IV can be used singly or as a mixture. 10 is H, an alkali metal ion, or a sugar residue, R 11 is H, an alkali metal ion, an organic base, a uronic acid, or a sugar residue. R 12 is H, a glucuronic acid and an alkali metal salt, an ammonium salt, or a sugar residue thereof. The sugar residue is independently one of a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide. The terpene glycoside of Formula IV can be used singly or as a mixture.

[0047] The terpene glycoside of Formula IV is preferably one or more of glycyrrhetic acid-3-O-glucuronide, glycyrrhizic acid, isoglycyrrhizic acid, glycyrrhetic acid, glycyrrhetic acid monoglucuronide, and sodium salt, ammonium salt, potassium salt of the above glycyrrhizic acid derivatives, and more preferably glycyrrhizic acid or ammonium glycyrrhizate.

[0048] The supramolecular nanocomplex, characterized in that the weight ratio of the flavonoid glycoside of Formula I to the colon / rectum low absorption active substance is in the range of A-B, wherein A is 1:0.0125 and B is 1:5; the total weight ratio of the flavonoid glycoside of Formula I to the terpene glycoside of Formula II and / or Formula III and / or Formula IV is in the range of C-D, wherein C is 1:0.25 and D is 1:5. There is no special limitation on the ratio of the terpene glycoside of Formula II, Formula III and Formula IV. The active substance in the supramolecular nanocomplex achieves high absorption in the colon / rectum.

[0049] The supramolecular nanocomplex, characterized in that it can further contain a surfactant, including one or more of the following: bile salts, vitamin E polyethylene glycol succinate (TPGS), sodium dodecyl sulfate (SLS), sodium docusate, lecithin, Tween 80, Tween 20, polyoxyethylene castor oil (EL35), polyoxyethylene hydrogenated castor oil, poloxamer, polyethylene glycol (15)-hydroxystearate, wherein the bile salts are one or more of the following: sodium taurocholate (STC), sodium glycocholate (SGC), sodium deoxycholate (SDC), sodium taurodeoxycholate (STDC), sodium glycodeoxycholate (SGDC), sodium glycochenodeoxycholate (SGCDC), sodium glycoursodeoxycholate (SGUDC), or free acid or potassium salt thereof. Preferably, one or both of sodium taurodeoxycholate (STDC) and sodium glycodeoxycholate (SGDC). The weight ratio of the flavonoid glycoside of Formula I to the surfactant is in the range of E-F, wherein E is 1:0.05 and F is 1:9.1.

[0050] The supramolecular nanocomplex, characterized in that it can further contain povidone and / or copovidone and / or polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus). The weight ratio of the flavonoid glycoside of Formula I to povidone and / or copovidone and / or polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus) is in the range of G-H, wherein G is 1:0.1 and H is 1:1.1.

[0051] The Zeta potential of the supermolecular nanocomplex is negative or the Zeta potential after in situ assembly with bile salts in the absorption site is negative.

[0052] A pharmaceutical composition comprising the above supermolecular nanocomplex, and a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient, such as excipient, diluent, binder, disintegrant, lubricant, flavoring agent, pH adjuster, osmotic pressure adjuster, thickening agent, plasticizer, colorant, film-forming agent, preservative, solvent, and combinations thereof.

[0053] The supermolecular nanocomplex or the pharmaceutical composition is characterized in that the supermolecular nanocomplex or the pharmaceutical composition is orally administered, and the supermolecular nanocomplex or the pharmaceutical composition can be released from the stomach, or from the duodenum, or from the jejunum, or from the ileum, or from the colon. The composition can be rapidly released or slowly released. The supermolecular nanocomplex or the pharmaceutical composition can be prepared into one or more of enteric or colon-soluble tablets, micro-tablets, capsules, pellets, micro-capsules, granules.

[0054] The supermolecular nanocomplex or the pharmaceutical composition is characterized in that the composition can be administered via the anus with or without a delivery device, and can be directly delivered to the colon or to the rectum; and can be prepared into an enema, a foam, a capsule, a tablet, a suppository, a gel, or an in-situ gel.

[0055] Advantages of the invention

[0056] Compared with the prior art, the supermolecular nanocomplex preparation has one or more of the following advantages:

[0057] (1) greatly improving the absorption of low-absorption substances in the colon / rectum, and facilitating the preparation of long-acting sustained-release preparations;

[0058] (2) improving patient compliance;

[0059] (3) improving drug exposure in the target organ and improving therapeutic efficacy;

[0060] (4) taking full advantage of the differences in microorganisms, enzymes, and transport proteins in the rectum / colon and other intestinal segments to prepare novel preparations different from ordinary release products, with differentiated pharmacokinetic characteristics, to improve the safety and effectiveness of drugs. BRIEF DESCRIPTION OF DRAWINGS

[0061] Figure 1. Comparison of pharmacokinetic curves of different formulations of APXB.

[0062] Figure 2. Cumulative release of F4 enteric capsules and reference formulations in simulated colon fluid.

[0063] Figure 3. Cumulative release of ticagrelor supramolecular nanocomplex F19-F21 in biorelevant media.

[0064] Figure 4. Cumulative release of ibutamautine supramolecular nanocomplex F22-F24 in biorelevant media.

[0065] Figure 5. Cumulative release of nimodipine supramolecular nanocomplex F25-F27 in biorelevant media.

[0066] Figure 6. F19-F27 supramolecular nanocomplex colon permeability coefficient.

[0067] Figure 7. F36-F37 finasteride supramolecular nanocomplex solution rectal dosing plasma, tissue exposure concentration.

[0068] Figure 8. F36-F37 finasteride supramolecular nanocomplex solution rectal dosing tissue / plasma exposure concentration ratio.

[0069] Figure 9. Apixaban, F14, F14-EB infrared spectrograms.

[0070] Figure 10 Apixaban, F14 DSC spectrograms.

[0071] Figure 11. Apixaban, F14 XRPD spectrograms. DETAILED DESCRIPTION

[0072] In order to make the objectives, technical solutions and advantages of the present application clearer, below, combined with specific examples and referring to the drawings, the present application is further described in detail.

[0073] In order to illustrate the features and advantages of the present application, the examples provide some experiments as illustrative examples, but the content of the present application is not limited to the examples. The materials and reagents used in the examples are all ordinary commercially available products.

[0074] The materials used in the present application, codes are as follows:

[0075] Table 4. Material name, role, code and source

[0076] The determination methods used in the present application are as follows:

[0077] (1) Nanoparticle size, PDI:

[0078] Measured by NanoBrook (model: 90Plus PALS) nanoparticle size analyzer. Before measurement, dilute the supramolecular nanocomplex solution to 100 μg / mL, take 50 μL and disperse into 1 mL deionized water, repeat 3 times for each sample.

[0079] (2) Zeta potential measurement:

[0080] The Zeta potential measurement was performed at room temperature 25℃, and the diffraction angle was set at 15℃. The supramolecular nanocomplex solution was diluted to 100 μg / mL before measurement, and 50 μL was dispersed in 1 mL of phosphate buffer (6.8) for measurement. Each sample was repeated 3 times.

[0081] (3) Active ingredient content determination:

[0082] The active ingredient content was determined by high performance liquid chromatography (HPLC) using an external standard method. The chromatographic conditions are shown in Table 5 below.

[0083] Table 5. Active ingredient content determination method

[0084] (4) Active ingredient related substance determination:

[0085] The related substance was detected by high performance liquid chromatography (HPLC). The chromatographic conditions are shown in Table 6 below.

[0086] Table 6. Active ingredient related substance determination method

[0087] (5) Active ingredient tissue sample detection method

[0088] Method for quantitatively detecting apixaban in rat plasma samples:

[0089] A single quadrupole liquid chromatography-mass spectrometry (LC-MS) instrument was used, and the chromatographic and mass spectrometric conditions are shown in Table 7 below:

[0090] Table 7. APXB rat plasma sample detection chromatographic conditions (LC-MS)

[0091] (6) Supramolecular nanocomplex infrared spectrum

[0092] 1) Test instrument: Spectrum 65 Fourier infrared spectrometer

[0093] 2) Test conditions: KBr tabletting method;

[0094] 3) Instrument calibration: According to the Chinese Pharmacopoeia 2020 edition four general rules 0402, polystyrene film was used for instrument calibration, and its spectrum was drawn, and the peaks at 3027 cm -1 , 2851 cm -1 , 1601 cm -1 , 1028 cm -1 , 907 cm-1 The instrument was calibrated with the absorbance peak of the instrument. The scanning range was 400-4000 cm -1 .

[0095] 4) Sample processing

[0096] The supermolecular nanocomplex solution was freeze-dried by freeze-drying method, and the blank carrier freeze-dried powder was prepared by the same process. The raw material drug was ground into a fine powder.

[0097] (7) X-ray powder diffraction (XRPD)

[0098] X-ray powder diffraction (XRPD) data was collected, and the instrument used was D / max-rB, and the test conditions were Cu Kα light source, 40 kV voltage, 40 mA current, slit DS1°, SS1°, RS0.3 mm, sampling step width: 0.02°, scanning speed: 10° / min, and the collection software was MDI Jade 5.0.

[0099] (8) Differential scanning calorimetry (DSC)

[0100] Differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) data were collected, and the instrument used was Mettler-TGA / DSC1 / 1100LF, and the analysis method parameters were temperature range: 30-350℃, scanning rate: 10℃ / min, protective gas: nitrogen, 50 ml / min. The collection software was STARe.

[0101] For example, apixaban, which has an apparent half-life of 12 hours, is theoretically possible to achieve once-daily administration. However, according to FDA review report CLINICAL PHARMACOLOGY AND BIOPHARMACEUTICS REVIEW(S) P98-100: Study CV185007 (Regional GI absorption), apixaban has little absorption in the distal small intestine and ascending colon, for example, when apixaban solution is administered, F 升结肠 = AUC 0-t结肠 / AUC 0-t口服 x 100% = 17.25%, and when the tablet is crushed and administered in the colon, F 升结肠 = AUC 0-t结肠 / AUC 0-t口 服 x 100% = 5.63%. Therefore, the solubility, permeability in the colon, and other factors caused the failure of the development of the apixaban sustained-release preparation of Pfizer / Bristol-Myers Squibb, and the final product on the market Apixaban tablet is administered twice a day, see patent CN102802608A. Therefore, drugs with low absorption in the small intestine and colon, even if the release transport time in the upper gastrointestinal tract is prolonged by existing technologies such as gastric floating and gastric retention, it is difficult to provide and maintain effective systemic exposure and exposure concentration during the drug administration interval, resulting in treatment or development failure.

[0102] Example 1. Supramolecular nanocomplexes F1-F8

[0103] A supramolecular nanocomplex was prepared using the drug with low absorption in the small intestine and colon, apixaban, as the active ingredient.

[0104] (1) Prescription composition:

[0105] Table 8. Prescription composition of supramolecular nanocomplexes F1-F8 (mg)

[0106] (2) Preparation process:

[0107] The carrier and surfactant in the prescription amount were added to 20 mL of pH 6.8 sodium phosphate buffer, heated to completely dissolve the carrier, cooled to room temperature, then 60 mL of acetone was added, stirred to mix evenly, then the prescription amount of APXB was added, heated to 55°C to completely dissolve the APXB, then 14 mL of purified water was added, stirred at 55°C for 1 hour, then the acetone was removed by rotary evaporator and concentrated to a certain volume, and then left to stand overnight. Centrifugation at 25°C / 15000 rpm for 5 min, the supernatant aqueous solution is the APXB supramolecular nanocomplex solution, and the apixaban content is detected by HPLC.

[0108] The above supramolecular nanocomplex solution was placed in a refrigerator at 2-8°C for 48 hours and at room temperature for 24 hours, both of which were clear.

[0109] According to the ELIQUIS (apixaban tablet) drug instruction (https: / / www.accessdata.fda.gov / drugsatfda_docs / label / 2021 / 202155s034lbl.pdf), the solubility of the pharmaceutical crystalline form of apixaban is 0.04 mg / mL within the physiological pH range (pH 1.2-7.4). The concentration (solubility) of apixaban in the supramolecular nanocomplexes F1-F8 prepared from apixaban can reach 7.95 mg / mL to 21.56 mg / mL, which is 199-539 times higher than that of the apixaban crystalline drug, thus successfully solving the problem of solubility, the first key factor for small intestine and colon absorption.

[0110] (3) Properties, particle size, PDI and zeta potential

[0111] Table 9. Properties, particle size, PDI and Zeta potential of the supramolecular nanocomplexes prepared in Example 1

[0112] According to the above results, the particle size of the supramolecular nanocomplexes prepared in Example 1 is between 6 nm and 17 nm, the solution is a clear and transparent solution, the PDI is not greater than 0.3, and the Zeta potential is negative.

[0113] Example 2. Supramolecular nanocomplexes F9-F18

[0114] (1) Prescription composition:

[0115] Table 10. Prescription composition of nanocomplexes F9-F18

[0116] (2) Preparation process:

[0117] According to the prescription, each component is taken, 10 mL of pH 6.8 sodium phosphate buffer is added for heating and dissolution, and after cooling to room temperature, 30 mL of acetone is added, stirred, APXB is added and heated to 55°C for dissolution, then 7 mL of water is added, stirred, and acetone is removed by rotary evaporation at 50°C. Centrifugation, collection of supernatant, preparation of apixaban supramolecular nanocomplex solution. HPLC detection of supramolecular nanocomplex content.

[0118] (3) Properties, particle size, PDI and Zeta potential

[0119] Table 11. Properties, particle size, PDI and Zeta potential of the supramolecular nanocomplexes prepared in Example 2

[0120] According to the above results, the particle size of the supramolecular nanocomplexes prepared in Example 2 is between 8 nm and 40 nm, the solution is a clear and transparent solution, the PDI is not greater than 0.3, and the Zeta potential is negative. After increasing the high molecular polymer in the prescription, the particle size increases, as shown in F14 to F18, and is positively correlated with the proportion of high molecular polymer in the prescription.

[0121] (4) In vitro colon tissue penetration evaluation

[0122] In vitro intestinal tissue was used for in vitro colon penetration evaluation; Franz diffusion cell method was used for determination. In order to investigate the influence of intestinal flora on carrier metabolism at the same time, the colon tissue of rats was taken, and the corresponding part of the colon contents was collected.

[0123] SD clean grade male rats were fasted overnight, and their body weight was measured (220 ± 10 g). The rectum was about 8 cm from the anus, and the colon was about 10 cm from the rectum. The colon segment was isolated and rinsed with 0.9% normal saline at 4°C. The colon was cut along the mesentery and washed with 4°C normal saline. Under a microscope, the colon segment was cut into 1 cm pieces, avoiding the parts with vesicle cells, and placed on the entrance of the receiving chamber of the Franz diffusion cell with the mucous layer facing up. The donor chamber was placed vertically on the receiving chamber, and the Franz diffusion cell was fixed with a clamp. The magnet was gently inserted into the receiving chamber through the sampling port, and 37°C pH 7.4 phosphate buffer was added as the receiving solution. The liquid level was about the scale line of the sampling port, and the bubbles were slowly released. The receiving chamber was checked for any leaks.

[0124] Comparative Example 1 (D1): The micronized apixaban was prepared into a suspension with 0.25% hydroxypropyl methylcellulose acetate succinate type H (HPMCAS H) in pH 7.4 phosphate buffer. The suspension was centrifuged, and the supernatant was collected as a control. The concentration of the supernatant was 0.42 mg / mL, as determined by HPLC.

[0125] Donor chamber:

[0126] Scheme A: The above-mentioned control, F1-F5, F7, F8, and F14 supramolecular nanocomplex solutions were used as the donor chamber solution.

[0127] Scheme B: 1 g of the collected colon contents was placed in a 10 mL sample tube, 8 mL of nitrogen-saturated pH 7.4 phosphate buffer was added, and the mixture was gently mixed. The control, F1-F3, F6, and F14 supramolecular nanocomplex solutions were diluted one-fold with the colon content suspension or paste to obtain the donor chamber components.

[0128] The circulating water bath temperature was set to 37°C, and the circulating water switch was turned on. After 5 minutes of incubation, the donor chamber was added with the prepared solution, and 3 replicates were set for each formula. The sampling time was 2 hr, 4 hr, 5 hr, 6 hr, and 7 hr.

[0129] The sampling volume was 500 μL, and the same volume of solution was added.

[0130] The sample solution was added with 215 μL of acetonitrile, vortexed for 30 seconds, and centrifuged at 3500 rpm for 10 minutes. The supernatant was collected, and the permeation amount of apixaban in the receiving solution was determined according to the apixaban content determination method.

[0131] The cumulative permeation amount Q (μg) was calculated according to the following formula, and plotted with time (hr) as the horizontal coordinate and Q as the vertical coordinate. The linear regression was performed on the three consecutive points with the largest slope, and the T was calculated by the linear regression equation. lag and J max or J ss The median time (hr) was plotted as the horizontal coordinate, and J as the vertical coordinate. The J after the J reached a steady state with time was taken as J max or J ss The apparent permeation coefficient (P app , cm·s -1 )

[0132] J = dQ / dt (μg / hr －1 ) (2) P app = J / (AC0) (3)

[0133] In the formula:

[0134] Q: cumulative permeation amount (μg)

[0135] Cn: APXB concentration in the receiving chamber at the nth sampling point, μg·mL -1 ;

[0136] J: absorption rate, μg / hr;

[0137] A: effective membrane area (0.19625 cm 2 );

[0138] C0: initial concentration of apixaban in the supply chamber (μg·mL －1 ).

[0139] The results are as follows:

[0140] Table 12. Absorption rate J and apparent permeation coefficient P app Results (N = 3, mean)

[0141] According to the results of the above in vitro colon tissue permeability experiment, the absorption rate of the comparative example 1 + colon content group was basically consistent with the results of the comparative example 1, and the colon content had no effect on the permeability of apixaban.

[0142] The colon content had no effect on the permeation rate and apparent permeation coefficient of apixaban in the F2 supramolecular nanocomplex; the apparent permeation coefficient of the F1 administration group was basically consistent with that of the comparative example 1, and the apparent permeation coefficient of the F1 + colon content group was about 4.61 times that of the comparative example 1; the apparent permeation coefficient of the F3 + colon content group was 1.10 x 10-5 cm·s -1 , which is 10.19 times that of Comparative Example 1 and 2.01 times that of the F3 administration group; the apparent permeation coefficients of the F4 and F8 administration groups are 6.21 and 9.72 times that of Comparative Example 1, respectively, which are significantly higher than that of Comparative Example 1.

[0143] The prescriptions of F14-F15 contain high molecular polymers in addition to flavonoid glycosides and terpene glycosides. The apparent permeation coefficients of the groups containing colonic contents of F14-F15 are significantly higher than those of the groups not containing colonic contents, and are significantly higher than that of Comparative Example 1.

[0144] (5) Rat rectal administration pharmacokinetic comparison

[0145] Test drugs: F1, F3, F4, F7, and F8 prescriptions in Example 1;

[0146] Control prescription: Take micronized apixaban drug substance, first add a small amount of dimethyl sulfoxide to dissolve, then add a pH 7.4 phosphate buffer containing 0.25% HPMCAS H to dissolve into a 10 mg / mL solution as a control sample.

[0147] Each rat was administered 12 mg / kg body weight according to the above prescriptions.

[0148] Experimental animals: Clean grade SD male rats, weighing 210-208 g, purchased from China Food and Drug Inspection Research Institute (Daxing), license number: SCKK(Jing)2022-0002. After purchasing, the rats were alternately adapted to the environment for one week, and were allowed to eat for more than 10 hours before administration, and were allowed to drink freely.

[0149] Before administration, each rat was intraperitoneally injected with 20% urethane 0.35 mL / 100 g, then a soft tube was inserted about 10 cm through the anus, and 0.3 mL of the test drug was administered through the soft tube, and after 30 min of observation without drug leakage, the rats were returned to the cage.

[0150] Sampling time points: Before administration and at 5 min, 30 min, 1 h, 2 h, 4 h, 6 h, 7 h, and 8 h after administration, about 0.5 mL of blood was taken from the inner canthus, EDTA-2K was used as an anticoagulant, 4500 rpm centrifugation for 10 min, plasma was taken, and stored in a -70°C refrigerator until sample detection.

[0151] Plasma sample pretreatment: Take 150 μL of plasma sample, add 10 μL of benadryl (BHLM) internal standard solution (600 ng / mL), vortex for 30 seconds, mix well, add 440 μL of acetonitrile, vortex for 30 seconds, stand for 10 min, centrifuge at 13000 rpm / 4°C for 10 min, take the supernatant for analysis;

[0152] The content of apixaban in plasma and tissue samples was analyzed by liquid chromatography-triple quadrupole mass spectrometry. The concentration of apixaban in plasma samples was calculated by internal standard method. According to the concentration data of apixaban at different time points, the pharmacokinetic parameters were calculated by Phoenix WinNonlin 7.0 to provide AUC 0-t , AUC 0-∞ , Cmax, Tmax and T 1 / 2 , etc. and their average values, and compared with the control. The results are shown in the following table, and the PK curve is shown in Figure 1.

[0153] Table 13. Pharmacokinetic results of colon administration in rats

[0154] From the above results, it can be seen that the nanocomposites F1, F3, F7 and F8 prepared in the examples can greatly improve the absorption of APXB in the colorectal site.

[0155] (6) Supramolecular nanocomplex solidification and redissolution

[0156] Select the supramolecular nanocomplex solutions of F1, F3, F4, F7, F8, F14 and F15 above, and place them in a Westlin bottle with a liquid layer thickness of not more than 0.5 cm. Place the bottle at room temperature in a-80℃ freeze dryer, pre-freeze for about 4 hours, and then freeze dry for more than 24 hours.

[0157] After freeze-drying of the above prescription, F1, F3, F4, F7 and F8 are white, crystalline block-shaped materials that disperse upon gentle touch of a glass rod; F14 and F15 have a foamy appearance after freeze-drying.

[0158] Weigh an appropriate amount of each prescription freeze-dried and finely powdered, and redissolve it in pH 7.4 phosphate buffer. Redissolve it according to the theoretical concentration of 5 mg / mL of apixaban, and detect the apixaban content, particle size, PDI and Zeta potential of the redissolved solution. The results are shown in the following table:

[0159] Table 14. Redissolution of supramolecular nanocomplex solution after solidification and its effect on particle size and PDI

[0160] According to the above results, the redissolution percentage of the supramolecular nanocomplex solution prepared according to Examples 1 or 2 after solidification is more than 85%, and the particle size and PDI are basically unchanged, indicating that the supramolecular nanocomplex prepared according to Examples 1 and 2 has good physical stability.

[0161] (7) Investigation of influencing factors

[0162] Place the sample of the example at a high temperature of 50℃, and take samples at 30 days. Detect the related substances by the method in Table 6. The results are as follows:

[0163] Table 15. Results of the influence factor impurity detection

[0164] From the above results, the chemical stability of the apixaban supramolecular nanocomplex lyophilized powder is good compared with the apixaban drug substance.

[0165] (8) Dissolution test in simulated colonic medium

[0166] Taking the F4 prescription as an example, an appropriate amount of F4 lyophilized powder (denoted as F4-FDP) was taken, and 3# or 0# enteric-coated capsules containing 2.5 mg and 5 mg of apixaban were prepared, respectively. At the same time, apixaban tablets were used as the reference preparation (RLD), with specifications of 2.5 mg / tablet and 5 mg / tablet, and were loaded into 3# or 0# enteric-coated capsules, respectively. The dissolution of the above apixaban preparations in simulated colonic fluid was investigated, with 3 parallel samples for each sample, and the average value was reported.

[0167] 1) Dissolution method: small cup method, paddle method, 75 rpm;

[0168] Dissolution medium: 37°C, simulated colonic fluid

[0169] Sampling time (min): 15, 30, 60, 90, 120, 180

[0170] Medium volume: 50 mL / cup

[0171] Sample pretreatment: 5 mL of sample was taken, centrifuged at 37°C / 14000 rpm for 5 min, 5 mL of blank medium at the same temperature was added, and after centrifugation, the sample was diluted with diluent. The content was detected by HPLC under the apixaban item in Table 5, and the cumulative dissolution was calculated. The results are shown in Figure 2.

[0172] From the cumulative dissolution results in the simulated colonic medium, the cumulative release of apixaban tablets 2.5 mg or 5 mg loaded into enteric-coated capsules was 57.4% and 35.6%, respectively, in 50 mL of simulated colonic fluid for 180 min. However, the F4 supramolecular nanocomplex lyophilized powder loaded into enteric-coated capsules, 2.5 mg and 5 mg, was nearly completely dissolved in 50 mL of simulated colonic fluid for 180 min.

[0173] ​Apixaban is just one case of straight / colon low absorption drugs, according to Connor A, King G, Jones K. Evaluation of human regional bioavailability to assess whether modified release development is feasible, https: / / cdn.technologynetworks.com / TN / Resources / PDF / 510.pdf 2014, review the results of 37 human regional drug absorption studies, about 65% of new chemical entities (NCEs) drugs and 53% of marketed drugs have relative bioavailability F 结肠 less than 60%; about 59% of new chemical entities (NCEs) drugs and 21% of marketed drugs have relative bioavailability F 结肠 less than 30%, the above technical solutions for solving apixaban are also applicable to other marketed and unmarketed straight / colon low absorption drugs.

[0174] Example 3. Supramolecular nanocomplex F19-F27

[0175] After oral administration of Ibrutinib (YBTN), severe liver first-pass metabolism leads to an oral absolute bioavailability of only 2.9%, and the maximum daily dose in the clinic is 420mg or 560mg depending on the indication, in addition, the first generation product Ibrutinib capsules have high intra-individual and inter-individual variability, and are prone to drug interactions with many drugs, causing great distress and safety risks for clinicians and patients.

[0176] Nimodipine (NMDP) is clinically used to prevent and treat ischemic neurological damage caused by cerebral vasospasm after aneurysmal subarachnoid hemorrhage. 30%-50% of patients with subarachnoid hemorrhage have difficulty swallowing, and 20%-30% are in a coma and cannot swallow drugs. Due to the solubility of nimodipine, the existing marketed products are ethanol-containing formulations regardless of injection or oral routes, in addition, due to the severe first-pass metabolism of nimodipine, its oral absolute bioavailability is 5%-15%. Nimodipine has a short elimination half-life, and needs to be taken four times a day, and the existing product has a "spring effect" in the blood concentration curve after oral administration, resulting in a high incidence of side effects of low blood pressure in patients, and the safety and compliance are very poor.

[0177] According to J Drug Assess. 2014 Aug 12; 3(1): 43-50, the metabolite ARC124910XX of Ticagrelor (TGRL) solution intravenous injection or suspension administered at different sites of the gastrointestinal tract has a significantly different proportion of plasma exposure (AUC 0-t ) of the prototype drug, and the proportions of intravenous injection, oral administration, proximal small intestine, distal small intestine and ascending colon are 13.9%, 39.1%,

[0178] 31.4%, 25.8% and 14.0%, respectively. The exposure of the metabolite ARC124910XX in the plasma of the ascending colon is equivalent to that of intravenous injection, and is significantly lower than that of oral administration, proximal and distal small intestine administration, indicating that colon administration can partially bypass the first-pass metabolism. However, if the drug itself has low absorption in the colon and rectum, and there is no corresponding technology to solve it, it will limit the realization of this goal. For example, the F 结肠 of Ticagrelor is 1119 / 3781x100%=29.6%, which limits the development of rectal / colon administration preparations.

[0179] The following are prepared into supramolecular nanocomposites with Ticagrelor, Ibutilide and Nimodipine as active ingredients, respectively.

[0180] (1) Prescription composition:

[0181] Table 16. Prescription composition of supramolecular nanocomposites F19-F27

[0182] (2) Preparation process:

[0183] 1) Ticagrelor

[0184] Weigh the prescription amount of each component, add water 10 mL to heat and dissolve, cool to room temperature, then add ethanol 30 mL, stir evenly, add active ingredient and heat to 55°C to dissolve, then add 7 mL of water, stir evenly, remove ethanol at 50°C using a rotary evaporator, stand overnight, centrifuge, and the supernatant is the supramolecular nanocomposite solution.

[0185] 2) Ibutilide

[0186] Weigh the prescription amount of each component, add water 10 mL to heat and dissolve, cool to room temperature, then add methanol-tetrahydrofuran (1:2, V / V) 25 mL, stir evenly, add active ingredient and heat to 55°C to dissolve, then add 5 mL of water, stir evenly, remove methanol and tetrahydrofuran at 50°C using a rotary evaporator, stand overnight, centrifuge, and the supernatant is the supramolecular nanocomposite solution.

[0187] 3) Nimodipine

[0188] Weigh the prescribed amount of each component, add 10 mL of water to heat and dissolve, cool to room temperature, then add the polymer, stir until completely dissolved, as the carrier solution.

[0189] Weigh 0.1 g of NMDP into 10 mL of ethanol, ultrasonic for 5 minutes to dissolve, as the drug-containing solution;

[0190] While stirring, slowly add the drug-containing solution to the carrier solution, continue stirring for 30 min after the addition is complete, remove the ethanol with a rotary evaporator at 45°C, stand overnight, centrifuge at 25°C / 15000 rpm for 5 min, and the supernatant is the nimodipine supramolecular nanocomplex solution.

[0191] The above supramolecular nanocomplexes were detected for the content of each active ingredient by HPLC.

[0192] (3) Characteristic evaluation

[0193] The results are shown in Table 17.

[0194] Table 17. Particle size, PDI and zeta potential detection results of F19-F27 in Example 3

[0195] The active ingredient structures and prescription compositions of the above supramolecular nanocomplexes are different, and the particle sizes of the prepared supramolecular nanocomplexes will have certain differences. Except for F27, the particle sizes of the supramolecular nanocomplexes prepared by other prescriptions are all less than 100 nm, and the particle size of F27 is 181.5 nm. The zeta potentials of all prescriptions are negative, and the PDI basically meets the requirements.

[0196] (4) Biological relevant dissolution experiment

[0197] 1) Experimental process

[0198] According to Journal of Controlled Release 125 (2008) 77-86, doi: 10.1016 / j.jconrel.2007.10.026, the intestinal flora of rats is closest to that of humans, and according to Scientific Reports | (2021) 11: 19929 | https: / / doi.org / 10.1038 / s41598-021-99379-6, except for the small intestine, the types of intestinal flora in the colon, rectum and cecum are basically the same, only the abundance is slightly different. Therefore, the contents of the rat rectum and colon were collected, mixed uniformly, and then added to the dissolution medium to investigate the dissolution of the above supramolecular nanocomplexes.

[0199] Before starting the dissolution experiment, the contents of the rat's straight and colon parts were collected, and the entire operation and experiment process was carried out in a nitrogen-filled glove box to provide an anaerobic environment for the intestinal flora. After the rat was anesthetized, the abdomen was quickly opened, the straight and colon were separated, and the contents in the straight and colon were collected, mixed, weighed, diluted into a paste according to 1 g of contents plus 10 mL of pH 7.0 nitrogen-filled phosphate buffer, and used, and an anaerobic environment was maintained.

[0200] Control sample:

[0201] Ticagrelor solution: an appropriate amount of ticagrelor fine powder was weighed, dissolved in pH 7.0 phosphate buffer-ethanol (7:2, V / V) containing 0.5% poloxamer 407 and 0.2% SGDC to prepare a solution containing about 5 mg of ticagrelor per 1 mL, which was used as a ticagrelor control solution;

[0202] Ibrutinib solution: an appropriate amount of ibrutinib drug substance was weighed, dissolved in pH 7.0 phosphate buffer containing 0.2% SGDC and 0.25% Soluplus to prepare a solution containing 3.5 mg of ibrutinib per 1 mL, which was used as an ibrutinib control solution;

[0203] Nimodipine control: commercially available nimodipine oral solution (60 mg: 10 mL) was used as a control.

[0204] 2 mL of the above F19-F27 prescription supramolecular nanocomposite solution and control solution were taken, 18 mL of dissolution medium containing the contents of the straight and colon were added, mixed uniformly, sealed, placed in a 37°C shaking bed for 3 hours, and 1 mL of sample was taken at 3, 6, 8, 10, 16, 20 hours after the start of shaking, while supplementing the same temperature and volume of dissolution medium. Each prescription was set up in triplicate.

[0205] 2) Sample pretreatment

[0206] The sample solution was added with 10 times the volume of 70% acetonitrile, vortexed for 30 seconds, and centrifuged at 15000 rpm for 10 minutes. The supernatant was injected according to the injection volume of the active ingredient content determination method, and the active ingredient content was determined, and the cumulative dissolution was calculated. The control sample solution was added with 5 times the volume of 70% acetonitrile, and the other processing steps were the same as the sample solution.

[0207] 3) The cumulative dissolution results are shown in Figures 3-5, respectively.

[0208] (1) From the dissolution curve results of ticagrelor, the cumulative dissolution of ticagrelor solution in the dissolution medium containing the contents of the small intestine and colon was about 12.24% in 24 hours, and the cumulative dissolution of the ticagrelor supramolecular nanocomplex prescriptions F19-F21 was 92.06%, 83.47% and 98.54%, respectively. The F21 prescription was completely released in 24 hours, and the F20 prescription was still in a slow release state although it was not completely released in 24 hours.

[0209] (2) From the dissolution curve results of ibritumomab, the cumulative dissolution of the ibritumomab solution (control) in the dissolution medium containing the contents of the small intestine and colon was about 18.31% in 24 hours, and showed a slow downward trend. The cumulative dissolution of the ibritumomab supramolecular nanocomplex prescriptions F22-F24 was 92.11%, 95.21% and 88.27%, respectively, and the cumulative release of each prescription was more than 85% in 24 hours, but still showed a slow release state.

[0210] (3) The control was a commercially available nimodipine oral solution containing ethanol. The cumulative release of the control in the dissolution medium containing the contents of the small intestine and colon was 17.31% in 24 hours. The cumulative release of each of the supramolecular nanocomplex prescriptions F25-F27 was more than 85% in 24 hours, among which the F26 prescription released the slowest, and the F27 prescription released the fastest although it had the largest particle size.

[0211] (5) Investigation of colon permeability

[0212] According to the "in vitro intestinal tissue permeability evaluation" procedure under Example 2, the rat colon tissue was isolated before the experiment, and the contents of the small intestine and colon were collected. The difference from the above was that the entire operation process was carried out in a nitrogen-filled glove box to ensure that the entire experimental process was in an anaerobic environment.

[0213] pH 7.0 phosphate buffer solution was filled with nitrogen;

[0214] The contents of the small intestine and colon were diluted into a paste by adding 8 mL of pH 7.0 phosphate buffer solution to 1 g, and the centrifuge tube was gently rotated to make the mixture uniform, serving as the supply chamber matrix.

[0215] The test solution was the supramolecular nanocomplex solution prepared according to the above control and F19-F27 prescriptions. About 3 mL of the above solution was taken and mixed with about 3 mL of the matrix containing the contents of the small intestine and colon to serve as the supply chamber components.

[0216] The receiving chamber was added with pH 7.4 phosphate buffer solution.

[0217] The temperature of the circulating water bath was set to 37°C, and after incubation for 5 minutes, the supply pool was added with the supply solution / content which had been incubated at 37°C, and 3 replicates were set for each prescription.

[0218] Sampling time points: 60 min, 90 min, 120 min, 150 min, 180 min;

[0219] Sampling volume: 500 μL, supplemented with an equal volume of isothermal solution.

[0220] The sample solution was added with 1-fold volume of acetonitrile solution, vortexed for 30 seconds, and centrifuged at 15000 rpm for 10 minutes. The supernatant was taken, and the content of each active ingredient was detected by HPLC according to the corresponding method in Table 5.

[0221] The permeation rate J and apparent permeation coefficient were calculated according to the formula in the "Evaluation of in vitro intestinal tissue permeability" under Example 2. The results are shown in Table 18 and Figure 6.

[0222] Table 18. J and apparent permeation coefficient P of control and F19-F27 supramolecular nanocomplexes app Results (N = 3, mean)

[0223] According to the permeability results, compared with the respective control, the supramolecular nanocomplexes can increase the apparent permeation coefficient by 2-5 times, as shown in the figure.

[0224] Example 4. Supramolecular nanocomplexes F28-F37

[0225] Flurbiprofen is the prodrug of flurbiprofen. The marketed dosage form is flurbiprofen fatty emulsion for injection, and the indications are postoperative and cancer analgesia. The current clinical administration route is intravenous injection. Although there are flurbiprofen oral sustained-release preparations on the market, due to the irritability of flurbiprofen to the gastrointestinal mucosa, many patients are unwilling to take them. Flurbiprofen ester is directly administered through the colon and / or rectum, and the expression of CES1 specific esterase in the colon / rectum is very low. After the drug is absorbed from the colon / rectum and enters the circulatory system, it is metabolized by esterase CES1 in the plasma or liver to flurbiprofen to exert its effect.

[0226] Kayfeng flurbiprofen ester fatty emulsion, 5 mL: 50 mg, after 50-fold dilution with normal saline for injection, the particle size was 195 nm.

[0227] (1) Prescription composition:

[0228] Table 19. Prescription composition of supramolecular nanocomplexes F28-F37

[0229] (2) Preparation process:

[0230] The components were taken according to the prescription, 10 mL of water was added to heat and dissolve, and after cooling to room temperature, 30 mL of ethanol was added, stirred uniformly, the active ingredient was dissolved, 3 mL of water was added, stirred uniformly, and the ethanol was removed by rotary evaporation at 50°C, and centrifugation was performed to obtain a nanocomposite solution. The content of the active ingredient was detected by HPLC.

[0231] The solubility of the active ingredient is greatly improved after being prepared into a supramolecular nanocomposite.

[0232] (3) Characteristic evaluation

[0233] Table 20. Particle size, PDI and zeta potential detection results of examples 4F28-F37

[0234] The above supramolecular nanosolution is still clear after being placed at room temperature or 2-8°C for 4 weeks.

[0235] (4) PK study of flurbiprofen supramolecular nanocomposite in rats by colon administration

[0236] 1) Experimental animals

[0237] SD male rats, weighing 240-251 g, were administered after being adapted to the environment for 12 hours of day-night alternation.

[0238] 2) Dosing regimen

[0239] All rats were fasted for more than 12 hours before administration, and all rats were randomly divided into four groups, with 5 rats in each group; the administration was performed according to the administration regimen in Table 18.

[0240] Table 21. Dosing regimen

[0241] Group A rats were injected with Kayven (i.e. flurbiprofen) diluted solution 0.25 mL through the tail vein;

[0242] The F28-F30 supramolecular nanocomposite solution was taken in an appropriate amount, diluted one time with pH 7.0 phosphate buffer, and administered into the colon of rats.

[0243] Group B, C, D colon administration: after the rats were anesthetized by inhaling ether, a silicone tube connected to a syringe was inserted through the anus (10 cm from the anus), and 0.25 mL of F28 or F29 or F30 flurbiprofen supramolecular nanosolution was poured into the colon, and then the silicone tube was slowly pulled out, the anus was pressed with hands and the rat's tail was lifted for 30 seconds.

[0244] 3) Bleeding points: 0.08, 0.25, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 8.0 and 16 hr after administration, 0.5 mL of blood was taken from the inner canthus, anticoagulated with sodium heparin, centrifuged at 3000 rpm for 10 min, and the plasma was separated for testing.

[0245] Pre-treatment of plasma samples: 200 uL of plasma sample was taken, 10 uL of internal standard solution (200 ng / mL of benadryl) and 590 uL of methanol were added, vortexed for 30 s, allowed to stand for 10 min, centrifuged at 4°C / 13000 rpm for 10 min, and the supernatant was injected for analysis.

[0246] Target analyte: flurbiprofen.

[0247] 4) Plasma sample analysis method

[0248] Table 22. Liquid chromatography and mass spectrometry conditions for detection of flurbiprofen in biological samples

[0249] Flurbiprofen was detected by internal standard method.

[0250] 3) Experimental results

[0251] Table 23. Pharmacokinetic parameters of flurbiprofen ester supramolecular nanocomplex administered to the colon of rats (n=5)

[0252] From the above results, it can be seen that after administration of flurbiprofen ester supramolecular nanocomplex to the colon, the absolute bioavailability of F28-F30 was 62.3%, 72.7% and 75.2%, respectively, and the elimination half-life of F28, F29 and F30 was about 1.6 times, 2.7 times and 2.7 times that of tail vein injection, respectively. The bioavailability was comparable to that of flurbiprofen sustained-release tablets, but the colon administration of the ester prodrug reduced the irritation of flurbiprofen to the intestinal mucosa, improved patient compliance and drug safety for patients who could not take flurbiprofen orally or tolerate flurbiprofen in the gastrointestinal tract, and reduced the risk of injection administration.

[0253] (5) Tissue distribution study of finasteride supramolecular nanosolution administered to the rectum of rats

[0254] The drugs for treating prostate cancer, cervical cancer, uterine cancer, ovarian cancer, ovarian polycystic, pelvic inflammatory disease, uterine fibroids, urethritis, bladder cancer and the like in clinic, due to the special location of the target organs of these diseases, the exposure of the drugs in the target organs is not high through the conventional oral or injection route of systemic administration, resulting in great systemic toxicity or other non-target organ toxicity, but the efficacy is very limited. For such drugs or diagnostic reagents for such diseases, the exposure of the drugs in the target organs of these diseases can be significantly improved by innovative delivery technology to solve the dissolution and release of the drugs in the rectum, improve the absorption of the drugs in this part, and further significantly improve the efficacy, safety or diagnostic accuracy, reduce the systemic exposure of toxic drugs or diagnostic reagents, and provide PK behavior matching the disease treatment or diagnosis.

[0255] For example, finasteride is widely used in clinic to treat symptomatic benign prostatic hyperplasia, reduce the risk of acute urinary retention, and reduce the risk of transurethral resection of the prostate and prostatectomy. In addition, finasteride is also often used by doctors off-label for female polycystic ovary syndrome. After oral administration of finasteride tablets, the gastrointestinal absorption is rapid, but the concentration of the drug in the treatment target organs / tissues is low, and the elimination is fast, which makes the treatment effect not obvious, and further increasing the systemic dose often causes systemic or other organ toxicity.

[0256] 1) Experimental animals

[0257] 7-week-old Sprague-Dawley male rats (body weight: 195-205 g) were used. The rats were fed for one week under the conditions of free water and feed, room temperature of 23±2℃ / RH 55±10%. The rats were fasted for more than 24 hours before administration, but not deprived of water, and the anal part was lightly touched to make them defecate. The rats were deprived of water 2 hours before administration. Water was restored 2 hours after administration, and food was given 4 hours after administration.

[0258] 2) Test drug:

[0259] Control: Finasteride suspension solution prepared with 0.5% methyl cellulose, concentration of 1 mg / mL;

[0260] Experimental drug: F36, F37 finasteride supramolecular nanocomplex solution, concentrations of 5.12 mg / mL and 4.99 mg / mL, respectively;

[0261] 3) Experimental animal grouping, administration scheme and administration method

[0262] The experimental animals were randomly divided into 3 groups, 12 rats in each group.

[0263] Table 24. Grouping of experimental animals

[0264] The dosing dose of all animals was 5 mg / kg body weight.

[0265] The oral administration group of rats was orally administered 1 mL of finasteride suspension by gavage, and then 1 mL of water.

[0266] Rectal administration: After the rats were anesthetized by inhaling ether, a silicone tube connected to a syringe was inserted into the anus (4 cm from the anus), 0.2 mL of F36 or F37 finasteride supramolecular nanosolution was injected into the rectum, and then the silicone tube was slowly pulled out, the anus was pressed with the hand, and the rat's tail was raised for 30 seconds.

[0267] 4) Sampling and biological sample pretreatment

[0268] Plasma and tissues: 3 rats per sampling point, at 0.5 hr, 1 hr, 6 hr, and 24 hr after administration, the rats were sacrificed after enucleation and blood collection (heparin anticoagulation), the rat's prostate, testis, epididymis, seminal vesicle, and bladder were quickly separated, rinsed with 4°C normal saline, the tissue surface was dried with a water-absorbing paper and weighed, the tissue was cut into small pieces, mixed with normal saline (4°C) at a mass ratio of 1:4, homogenized with a homogenizer, frozen and thawed in liquid nitrogen three times, centrifuged at 4°C and 13000 rpm for 10 minutes, and the homogenate was used as the tissue homogenate.

[0269] Plasma: 200 μL of plasma sample and 100 μL of betamethasone internal standard solution (5 ng / mL) were placed in a 4 mL centrifuge tube, vortexed for 30 seconds, 50 μL of 0.1M sodium hydroxide solution and 1.5 mL of extraction solvent (ethyl acetate-n-hexane, 1:1, V / V) were added, vortexed for another 10 seconds, centrifuged at 4°C and 3000 rpm for 1 min, frozen on dry ice, the organic phase was separated and placed in a clean centrifuge tube, the sample was blown dry with nitrogen flow at 40°C, 100 μL of diluent (acetonitrile-0.1% formic acid water, 90:10, V / V) was added for reconstitution, vortexed for 30 seconds, and 20 μL of supernatant was analyzed by LC / MS / MS.

[0270] Tissue sample processing: 200 μL of tissue homogenate was taken, 100 μL of internal standard solution was added, vortexed for 30 seconds, 50 μL of 0.1M sodium hydroxide solution and 1.5 mL of extraction solvent (ethyl acetate-n-hexane, 1:1, V / V) were added, vortexed for another 10 seconds, centrifuged at 4°C and 13000 rpm for 3 min, frozen on dry ice, the organic phase was separated and placed in a clean centrifuge tube, the sample was blown dry with nitrogen flow at 40°C, 100 μL of diluent (acetonitrile-0.1% formic acid water, 90:10, V / V) was added for reconstitution, vortexed for 30 seconds, and 20 μL of supernatant was analyzed by LC / MS / MS.

[0271] The above tissue samples can be further diluted with diluent according to the measured concentration.

[0272] 5) Detection method of finasteride in biological samples

[0273] The content of finasteride in plasma and tissue samples was analyzed by liquid chromatography-triple quadrupole mass spectrometry. The liquid chromatography and mass spectrometry conditions are shown in Table 25.

[0274] Table 25. Liquid chromatography and mass spectrometry conditions for detection of finasteride in biological samples

[0275] The concentration of finasteride in plasma and different tissue samples was detected by internal standard method and compared with the control group.

[0276] 6) Test results

[0277] From the results of animal tissue distribution, it can be seen that after oral gavage administration of the control solution, the plasma and each tissue rapidly reached the peak at about 1 hour, and the drug detection level in most tissues at 6 hours was already very low, and the concentration at 24 hours was close to the detection limit. After rectal administration of F36 and F37 supramolecular nanosolutions, due to the action of microbial enzymes in the rectum, the drug was slowly released and absorbed, so the concentration of F36 and F37 in the plasma and tissues at 6 hours and 24 hours after administration was still significantly higher than that of the control group, which was very critical for maintaining drug efficacy.

[0278] According to FIGS. 7-8, the concentration ratio of F36-F37 in tissues / plasma after administration was 2-8 times higher than that of the control group, and the ratio in the prostate was the highest.

[0279] The exposure level of finasteride in different tissues / organs after rectal administration of the drug can be known, and compared with systemic administration, rectal administration can significantly improve the exposure of the drug in target organs / tissues, and by optimizing the administration dose, the systemic exposure can be significantly reduced, the toxicity to other non-target organs can be reduced, and similar drugs or diagnostic reagents for treating other diseases are also applicable.

[0280] Example 5. Supramolecular nanocomplexes F38-F43

[0281] Tacrolimus has a high immunosuppressive effect for kidney transplant patients, but its systemic administration causes toxic side effects, premature failure of the transplanted organ, complications, a narrow therapeutic window, drug interactions, high PK variability within and between individuals, and the need for frequent monitoring of blood drug concentrations during treatment, which is a huge clinical pain point for doctors and patients. According to Journal of Pharmacological Sciences 139 (2019) 65-71, a model animal of cynomolgus monkeys was used to study the results of intramuscular injection or oral administration of tacrolimus. The results showed that the intramuscular injection of tacrolimus had significantly lower PK variability within and between individuals than oral administration; the Ctrough, Cmax and AUC variability after repeated intramuscular administration were also significantly lower than oral administration, indicating that the first-pass metabolism caused by digestive enzymes, drug solubility, food effect, drug interactions, etc. are the main reasons for the high variability of tacrolimus by oral administration.

[0282] In addition to tacrolimus, other drugs for treating autoimmune diseases also have the above-mentioned pain points. Traditional intravenous injection or oral administration of immunosuppressive agents such as cyclosporine, sirolimus, budesonide, methotrexate, and glucocorticoids also have the above-mentioned problems.

[0283] The present application takes cyclosporine, tacrolimus and sirolimus in calcineurin inhibitors as an example to prepare a supramolecular nanocomposite.

[0284] (1) Prescription and preparation process

[0285] Table 26. Prescription composition and content results of F38-F43 in Example 5

[0286] Preparation process: according to the prescription amount of active ingredients, flavonoid glycosides, terpene glycosides, high molecular polymers or surfactants, add 20 mL of ethanol, then add 15 mL of water for injection, heat to 60°C to dissolve, stir for 1 hour, remove ethanol with a 50°C rotary evaporator, and concentrate to about 10 mL. Let stand overnight at room temperature, centrifuge at 15000 rpm for 10 minutes, and the supramolecular nanocomposite is obtained. F38 is a clear liquid at room temperature, and becomes a gel after being placed at 2-8°C for 48 hours, and becomes a solution again after being restored to room temperature. The rest of the prescriptions do not change in nature whether they are placed at room temperature or 2-8°C for two weeks.

[0287] (2) Characteristic evaluation

[0288] Table 27. Particle size, PDI and Zeta potential detection results of F38-F43

[0289] The above-mentioned supramolecular nanocomposite solution is still clear after being placed at room temperature or 2-8°C for 2 weeks.

[0290] (3) Rat pharmacokinetic comparison

[0291] Test drug: F43 supramolecular nanocomplex solution was taken, and was freeze-dried in a freeze dryer for 48 hours to obtain a freeze-dried powder. The control was sirolimus raw material.

[0292] Animals: Wistar rats, 8 weeks old, male, body weight 180-220 g.

[0293] Twenty rats were randomly divided into two groups (T1, T2), 10 rats in each group, and were fasted for more than 12 hours before administration, and were allowed to drink water freely, and the water supply was stopped 2 hours before administration.

[0294] Dose: Sirolimus tablets for human oral administration, once a day (1 mg / day, about 0.0143 mg / kg). The equivalent dose of rats and humans is 0.09 mg / kg, but when administered at the equivalent dose, the blood concentration is too low to be detected, so the dose is increased by 50 times, and administered at 4.5 mg / kg. The amount of administration was calculated according to the body weight of the rats, and the F43 freeze-dried powder and sirolimus raw material were respectively loaded into No. 9 capsules.

[0295] Administration method: Sirolimus raw material capsules were administered directly by using a rat oral gavage device, and 1 mL of water was given after administration. Two hours after administration, the rats were allowed to drink water freely. The F43 freeze-dried powder capsules were delivered to the colon site through the anus using a dosing device, and the dosing device was slowly pulled out.

[0296] Blood, 0.5 mL, was taken from the inner canthus at 0.5, 1, 1.5, 2, 3, 4, 5, 7, 9, 12 and 24 hours after administration, and 3% heparin sodium was used as an anticoagulant. Centrifugation at 4°C / 3000 rpm for 10 minutes, separation of plasma, and storage in a -20°C refrigerator.

[0297] The content of sirolimus in the plasma was determined by LC / MS / MS detection (see above).

[0298] Table 28. Sirolimus rat pharmacokinetic results

[0299] According to the above results, the bioavailability of sirolimus supramolecular nanocomplex colon administration is greatly improved compared with oral administration of raw material.

[0300] The above prepared supramolecular nanocomplex solution can be directly prepared into different pharmaceutical composition preparations according to the needs of treating diseases, or can be freeze-dried, and the obtained freeze-dried powder is added to other pharmaceutically acceptable excipients to prepare a preparation.

[0301] The administration route can be oral or rectal administration. For oral administration, such as oral enteric preparations include immediate-release preparations and sustained-release preparations: enteric capsules, enteric granules, enteric pellets, enteric tablets, etc.; oral colon-specific preparations such as colon-specific capsules, colon-specific pellets, colon-specific tablets, etc.; enema, suppository, rectal / colon gel, rectal / colon foam, etc. for rectal administration.

[0302] Example 6. Supramolecular nanocomplex F44-66 and comparative examples D2-D7

[0303] (1) F44-F52

[0304] 1) Supramolecular nanocomplex prescription:

[0305] Table 29. Supramolecular nanocomplex prescription of F44-F52

[0306] 2) Preparation process:

[0307] F44-F52: Dissolve the prescribed amount of surfactant and cyclosporine in an appropriate amount of ethanol, and prepare for use. Add water or pH buffer solution 14 mL to the prescribed amount of carrier and high molecular polymer, heat to dissolve, cool to room temperature, adjust the pH to the target pH, slowly add the aqueous phase to the organic phase while stirring, continue to stir at room temperature for 1 hour, recover ethanol at 50°C under reduced pressure to 10 mL, hydrate at 37°C / 250 rpm for 0.5 hours, centrifuge at room temperature / 13000 rpm for 10 minutes, and take the supernatant to detect the content after standing for 24 hours.

[0308] (2) F53-F58 and D2-D4

[0309] 1) Supramolecular nanocomplex and D2-D4 and F53-F58 prescription:

[0310] Table 30. Supramolecular nanocomplex prescription of D2-D4, F53-F58

[0311] 2) Preparation process:

[0312] F53-F58: Dissolve the prescribed amount of surfactant and cyclosporine in an appropriate amount of ethanol, and prepare for use. Add water or pH buffer solution 10 mL to the prescribed amount of carrier and high molecular polymer, heat to dissolve, cool to room temperature, adjust the pH to the target pH, slowly add the aqueous phase to the organic phase while stirring, continue to stir at room temperature for 1 hour, evaporate at 50°C under reduced pressure, hydrate at 37°C / 250 rpm for 0.5 hours, centrifuge at room temperature / 13000 rpm for 10 minutes, and take the supernatant to detect the content after standing for 24 hours.

[0313] 3) Results and discussion

[0314] According to the comparative examples D2, D3, D4 and examples F53, it can be seen that the content of the nanocomposites prepared by terpene glycosides alone or terpene glycosides + high molecular excipients, flavone glycosides alone is very low, while the measured content of the supramolecular nanocomposites prepared by the prescriptions F53-F55 used in combination is basically consistent with the theoretical feeding amount, close to complete encapsulation. After increasing the theoretical feeding amount of the active ingredients, the encapsulation rate of the supramolecular nanocomposites prepared by the prescription F56 is also close to 85%, and the encapsulation rate of the prescriptions F57-F58 is close to 100%, i.e. 19.37 mg / mL and 10 mg / mL. Compared with the comparative examples D2-D4, the content of the prepared supramolecular nanocomposite solution is increased by about 28 times.

[0315] (3) F59-F68 and D5-D7

[0316] 1) Supramolecular nanocomposites and prescriptions D5-D7:

[0317] Table 31. Prescriptions of supramolecular nanocomposites D5-D7, F59-F66 Note: * is the result of detection under the condition of storage at 2-8°C for 14 days.

[0318] 2) Preparation process:

[0319] Dissolve tacrolimus and surfactant in an appropriate amount of ethanol solution.

[0320] Weigh the prescribed amount of carrier and high molecular polymer, add 14 mL of phosphate buffer solution with pH 5.3, mix well, and drop the tacrolimus-containing ethanol solution into the above-mentioned carrier-containing solution while stirring. Stir magnetically at room temperature for 1 hour. Remove ethanol to 10 mL at 50°C under reduced pressure, hydrate at 37°C / 250 rpm for 0.5-1 hour, and centrifuge at room temperature / 13000 rpm for 10 minutes. Take the supernatant.

[0321] Use the method of Table 5 to detect the content by HPLC, and respectively place at room temperature or 2-8°C to observe the physical stability. The content detection results of prescriptions F59, F60, F61, F62, F63, F64, F67, F68 and D7 stored at 2-8°C for 14 days are basically consistent with those of 0 days, and the solutions are still clear and uniform; D7 has a large amount of precipitation, and the content detection result after 14 days is 4.21 mg / mL.

[0322] (4) Particle size, PDI and potential

[0323] Table 32. Particle size, PDI and zeta potential detection results of F44-F68 and D2-D7

[0324] Example 7. Preparation of compositions

[0325] The following are examples of several pharmaceutical composition preparations:

[0326] (1) Preparation of enema

[0327] F16C: Prepare 10 mL of the supramolecular nanocomplex solution according to the prescription of F16, dilute with purified water to 100 mL, filter with 0.45 μm and 0.22 μm filter membranes, and aliquot to obtain an enema of 1.2 mg / mL.

[0328] F24C: Prepare 100 mL of the supramolecular nanocomplex solution according to the prescription of F24, filter with 0.45 μm and 0.22 μm filter membranes, and aliquot to obtain an enema of 33 mg / mL.

[0329] (2) Preparation of gel

[0330] F43C: Prepare the supramolecular nanocomplex solution according to the prescription of F43, and add purified water to prepare a 5 mg / mL solution for standby; take an appropriate amount of sodium hyaluronate, add purified water to prepare a 6% gel for standby; take the above solution and sodium hyaluronate gel, mix them uniformly at a ratio of 1:1 (V / V), aliquot, and prepare a 0.25% gel.

[0331] (3) Preparation of foam

[0332] F41C: Prepare 20 mL of the supramolecular nanocomplex solution according to the prescription of F41, dilute with purified water to 100 mL, pour into a pressure tank, flush into a propellant such as HFA-134a, isobutane, etc., and install a quantitative nozzle to obtain a foam.

[0333] (4) Preparation of enteric / colonic tablets

[0334] Table 33. Supramolecular nanocomplex F69-F71 prescription composition (mg)

[0335] The carrier and surfactant in the prescribed amount were added to 20 mL of sodium phosphate buffer at pH 6.8, and the carrier was completely dissolved by heating. After cooling to room temperature, 60 mL of acetone was added, and the mixture was stirred until uniform. The APXB in the prescribed amount was added, and the APXB was completely dissolved by heating to 55°C. Then, 14 mL of purified water was added, and the mixture was stirred at 55°C for 1 hour. The acetone was removed by a rotary evaporator, and the volume was concentrated. The aqueous solution was collected, centrifuged at 25°C / 15000 rpm for 5 min, and the supernatant was freeze-dried and finely ground to obtain the freeze-dried powder of the APXB supramolecular nanocomplex. The content of the freeze-dried powder was detected, and the content of F69 was 12.1%, the content of F70 was 11.7%, and the content of F71 was 11.6%. The APXB supramolecular nanocomplex solution was prepared according to the prescription of F1, freeze-dried, finely ground, and the content of the freeze-dried powder was detected, which was 10.2% (w / w).

[0336] The TGRL supramolecular nanocomplex solution was prepared according to the prescription of F19, freeze-dried, finely ground, and the content of the freeze-dried powder was detected, which was 11.8% (w / w).

[0337] The YBTN supramolecular nanocomplex solution was prepared according to the prescription of F23, freeze-dried, finely ground, and the content of the freeze-dried powder was detected, which was 12.4% (w / w).

[0338] The above freeze-dried powder was taken and tablets were prepared according to the following prescription:

[0339] Table 34. Tablet prescription of F1, F19, F23, F69-F70 supramolecular complex

[0340] F1 ET, F1 CT preparation process: The F1 nanocomplex freeze-dried powder, microcrystalline cellulose PH102, and cross-linked polyvinylpyrrolidone were mixed uniformly, and then magnesium stearate was added and mixed uniformly. The tablets were compressed using a φ7 mm round punch, and the tablet weight was 130 mg. The tablet hardness was controlled to be 30-60 N, and the disintegration time was 0.5-5 min. The above tablets were coated with an enteric or colon soluble pre-coating agent, respectively, to obtain F1 enteric tablets (ET) and F1 colon soluble tablets (CT).

[0341] F19 CT: The F19 nanocomplex freeze-dried powder, microcrystalline cellulose PH101, and half of the prescribed amount of cross-linked polyvinylpyrrolidone were mixed uniformly, and then half of the prescribed amount of magnesium stearate was added and mixed uniformly. The mixture was dry granulated, and the remaining cross-linked polyvinylpyrrolidone and magnesium stearate were added and mixed uniformly. The tablets were compressed using a 12 mm round punch, and the tablet weight was 450 mg. The tablet hardness was controlled to be 60-120 N, and the disintegration time was 0.5-5 min. The above tablets were coated with a colon soluble pre-coating agent to obtain F19 CT colon soluble tablets.

[0342] F23CT: Take F23 nanocomposite lyophilized powder, microcrystalline cellulose PH101, 1 / 2 prescription amount of crosslinked polyvidone, mix evenly, then add 1 / 2 prescription amount of magnesium stearate, mix evenly, dry granulation, add the remaining crosslinked polyvidone and magnesium stearate, mix evenly, use 16*9mm capsule-shaped punch to press tablets, tablet weight 510mg, control tablet hardness 60-120N, disintegration time 0.5-5min. Take the above tablets, coat with a colon-specific pre-mixed coating agent, and F23CT colon-specific tablets are obtained.

[0343] F69CT, F70CT, F71CT: Take F69, F70, F71 nanocomposite lyophilized powder, microcrystalline cellulose PH102, crosslinked polyvidone, mix evenly, then add magnesium stearate, mix evenly, respectively use φ9mm, φ7mm, φ7mm round punch to press tablets, tablet weight 210mg, 130mg, 130mg, respectively control tablet hardness 60-120N, 30-60N, 30-60N, disintegration time 0.5-5min. Take the above tablets, respectively coat with a colon-specific pre-mixed coating agent, and F69CT, F70CT, F71CT are obtained.

[0344] (5) Preparation of colon-specific capsules

[0345] Prepare APXB supramolecular nanocomposite solution according to the F17 prescription, freeze-dry, and grind finely to obtain nanocomposite lyophilized powder. The content of the lyophilized powder is 10.1% (w / w).

[0346] Prepare NMDP supramolecular nanocomposite solution according to the F25 prescription, freeze-dry, and grind finely to obtain nanocomposite lyophilized powder. The content of the lyophilized powder is 8.6% (w / w).

[0347] Prepare sirolimus supramolecular nanocomposite solution according to the F42 prescription, freeze-dry, and grind finely to obtain nanocomposite lyophilized powder. The content of the lyophilized powder is 6.6% (w / w).

[0348] Take the nanocomposite lyophilized powder and prepare capsules according to the following prescription:

[0349] Table 35. Capsule prescription of F17, F25, F42 supramolecular nanocomposites

[0350] F17CC preparation process: Take F17 nanocomposite lyophilized powder, microcrystalline cellulose PH101, crosslinked polyvidone, 2 / 3 prescription amount of magnesium stearate, mix evenly, dry granulation, then add silicon dioxide and the remaining magnesium stearate, mix evenly, fill 0# colon-specific hollow capsules with a capsule filling machine, control the content loading to be in the range of 300mg-320mg, and pack in HDPE bottles to obtain colon-specific capsules (CC).

[0351] F25CC Preparation Process: Take F25 nanocomposite lyophilized powder, microcrystalline cellulose PH101, croscarmellose sodium, 2 / 3 of the prescription amount of magnesium stearate, mix uniformly, dry granulation, then add silicon dioxide, the remaining magnesium stearate, mix uniformly, fill into 0# colon-void capsules with capsule filling machine, control the content loading within the range of 290mg-310mg, fill into HDPE bottles, and the colon-void capsules are obtained.

[0352] F42CC Preparation Process: Take F42 nanocomposite lyophilized powder, microcrystalline cellulose PH101, crospovidone, 2 / 3 of the prescription amount of magnesium stearate, mix uniformly, dry granulation, then add silicon dioxide, the remaining magnesium stearate, mix uniformly, fill into 3# colon-void capsules with capsule filling machine, control the content loading within the range of 105mg-115mg, aluminum plastic packaging, and the colon-void capsules are obtained.

[0353] Experimental Example 8. Infrared spectrum, DSC and XRPD characterization of supramolecular nanocomposites

[0354] F14 blank supramolecular nanocomposites (without apixaban, denoted as F14-EB) were prepared according to the F14 prescription in Table 7 (without apixaban), and the preparation process was completely consistent with that of the F14 prescription with drug. The particle size, PDI and Zeta potential were detected, and the lyophilization was performed using the same lyophilization process as the F14 prescription.

[0355] The apixaban bulk drug powder was finely ground.

[0356] (1) Infrared spectrum analysis

[0357] The F14 lyophilized powder, F14-EB and apixaban bulk drug were subjected to infrared spectrum analysis, respectively, as shown in Figure 9.

[0358] The particle size detection results of F14-EB were 22.6±0.3nm, PDI was 0.279±0.003, and Zeta potential was -8.78±0.89mV.

[0359] According to the infrared spectrum of apixaban bulk drug and the infrared spectrum of F14 supramolecular nanocomposites, since apixaban is wrapped in the middle of the carrier, most of the characteristic peaks of apixaban are covered by the carrier peaks. According to the infrared spectra of F14-EB and F14, after drug loading, the following characteristic peaks from the bulk drug appear in the infrared spectrum of F14, such as 607.12cm -1 , 704.42cm -1 , 757.64cm -1 , while the characteristic peaks of 1493.8cm -1 , 1710.78cm -1 in the infrared spectrum of F14-EB disappear; 1246.74cm -1 , 1370.38cm-1 and 1658.99 cm -1 Characteristic peak shift +4 cm in F14 infrared spectrum -1 .

[0360] (2) DSC analysis

[0361] According to the DSC test results of APXB bulk drug, APXB bulk drug has an endothermic peak at 243.6℃, while the endothermic peak of F14 supramolecular nanocomplex at this position disappears, and has a relatively wide endothermic peak at 76.9℃. The DSC spectra of APXB bulk drug and supramolecular nanocomplex are shown in Figure 10.

[0362] (3) XRPD analysis

[0363] The XRPD spectrum of APXB bulk drug presents characteristic diffraction peaks of crystalline drugs, while the XRPD spectrum of F14 supramolecular nanocomplex has no obvious characteristic diffraction peaks. The XRPD spectra of APXB bulk drug and supramolecular nanocomplex are shown in Figure 11.

[0364] The above specific embodiments further illustrate the purpose, technical solutions and beneficial effects of the present application. It should be understood that the above description is only for specific embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. Supramolecular nanocomplexes, characterized in that, The nanocomplex comprises an active substance and a carrier, the carrier comprising flavonoid glycoside and terpene glycoside, wherein the bioavailability of the oral formulation after oral administration compared to the bioavailability of the oral formulation after oral administration of the active substance is the ratio F of the bioavailability after colonic or rectal administration to the bioavailability after oral administration 结肠 or F 直肠 ≤ 60 %, wherein F 结肠 = AUC 结肠 / AUC 口服 x 100 %; F 直肠 = AUC 直肠 / AUC 口服 x 100 %, preferably the active substance is a drug, a diagnostic agent or a nutritional supplement, wherein the bioavailability is expressed as the area under the plasma or serum or whole blood concentration-time curve AUC of the active substance.

2. The supramolecular nanocomplex of claim 1, wherein, wherein the ratio F of the bioavailability after colonic or rectal administration of the oral formulation to the bioavailability after oral administration of the oral formulation compared to the bioavailability after oral administration of the active substance 结肠 or F 直肠 ≤ 40%.

3. The supramolecular nanocomplex of claim 1 or 2, wherein, The Zeta potential of the supramolecular nanocomplex is negative or is negative after in situ assembly with bile salts in the absorption site.

4. The supramolecular nanocomplex according to any one of claims 1 to 3, characterized in that, The active substance is one or more of apixaban, rivaroxaban, edoxaban, ticagrelor, dabigatran, carvedilol, zafirlukast, cyclosporine, nitrendipine, gabapentin, pregabalin, cannabidiol, acipimox, tacrolimus, sirolimus, metformin, imeglimin, afatinib, bosutinib, colchicine, everolimus, ledipasvir, naloxegol, budesonide, bromocriptine, haloperidol, morphine, pyridostigmine, naloxone, naltrexone, thioridazine, selegiline, tacrine, terbutaline, lidocaine, pazopanib, prucalopride, rifaximin, ranolazine, furosemide, ritonavir, baclofen, captopril, benazepril, sumatriptan, cimetidine, ranitidine, fexofenadine, atenolol, ciprofloxacin, silodosin, prostat, acrivastine, ibutilide, simvastatin, lovastatin, pravastatin, atorvastatin, rosuvastatin, halofuginol, cinnarizine, griseofulvin, paclitaxel, docetaxel, topotecan, nimodipine, nisoldipine, nicardipine, fenofibrate, mycophenolate mofetil, mycophenolic acid, fenofibrate ester, chlorphenesin, probucol, tamsulosin, alfuzosin, finasteride, alendronate sodium, clodronate sodium, etidronate sodium, pamidronate sodium, risedronate sodium, zoledronate sodium.

5. The supramolecular nanocomplex of any one of claims 1-4, wherein, The flavonoid glycoside is a flavonoid glycoside of the structure of Formula I. R1 is a sugar residue, being one of monosaccharide, disaccharide, oligosaccharide, polysaccharide; R2, R3, R4 are one of H, OH, OR5; R5 is one of C1-C10 alkane or is one of monosaccharide, disaccharide, oligosaccharide, polysaccharide. terpene glycoside is one or more of a terpene glycoside of structure II, a terpene glycoside of structure III, a terpene glycoside of structure IV, R6 is hydroxyl, sugar residue, R7 is H, methyl, ethyl, isopropyl, alkali metal ion or sugar residue; R8, R9 are H or sugar residue, and are not H at the same time in the terpene glycoside of formula III; R 10 is H, alkali metal ion or sugar residue, R 11 is H, alkali metal ion, organic base, sugar acid, sugar residue, R 12 is H, glucuronic acid and its alkali metal salt, ammonium salt or sugar residue; R6, R7, R8, R9, R 10 , R 11、 R 12 The sugar residue is independently one of monosaccharide, disaccharide, oligosaccharide and polysaccharide, The terpene glycoside in the supramolecular nanocomplex is a single compound of the terpene glycoside shown in the structures of formula II, formula III and formula IV, or is a mixture of the terpene glycoside shown in the structures of formula II, formula III and formula IV.

6. The supramolecular nanocomplex of claim 5, wherein, the flavonoid glycoside of Formula I is one or more of Naringin dihydrochalcone, Neo-Hesperidin dihydrochalcone, Phloridzin, Rutin, Aspartame, 1-(3-beta-D-glucopyranosyl-2,4,6-trihydroxyphenyl)-3-(4-hydroxyphenyl)-1-propanone; the terpene glycoside of Formula II is one or more of Stevioside, Rebaudioside A, Rebaudioside B and salts thereof, Rebaudioside C, Rebaudioside D, Rebaudioside E, Rebaudioside F, Rebaudioside M, Rebaudioside N, Rebaudioside O, Dulcoside A, Dulcoside B, Rubusoside, Steviol, Steviolbioside and salts thereof; the terpene glycoside of Formula III is one or more of Mogroside V, Mogroside IV, Mogroside IIA1, Mogroside IIA2, Mogroside VIA, Mogroside VIB, Mogroside III, Mogroside I, Siamenoside I; the terpene glycoside of Formula IV is one or more of Glycyrrhizic acid, Sodium salt of Glycyrrhizic acid, Ammonium salt of Glycyrrhizic acid, Potassium salt of Glycyrrhizic acid.

7. The supramolecular nanocomplex of any one of claims 1-6, wherein, the flavonoid glycoside of Formula I is in a weight ratio to the active substance in the range of A-B, wherein A is 1:0.0125 and B is 1:5; the flavonoid glycoside of Formula I is in a total weight ratio to the terpene glycoside represented by Formula II and / or Formula III and / or Formula IV in the range of C-D, wherein C is 1:0.25 and D is 1:

5.

8. The supramolecular nanocomplex of any one of claims 1-7, wherein the supramolecular nanocomplex further comprises a surfactant selected from one or more of a bile salt, Vitamin E polyethylene glycol succinate (TPGS), sodium lauryl sulfate (SLS), sodium docusate, lecithin, Tween 80, Tween 20, polyoxyethylene castor oil (EL35), polyoxyethylene hydrogenated castor oil (Rh40), poloxamer, polyethylene glycol (15)-hydroxystearate, wherein the bile salt is one or more of Sodium taurocholate (STC), Sodium glycocholate (SGC), Sodium deoxycholate (SDC), Sodium taurodeoxycholate (STDC), Sodium glycodeoxycholate (SGDC), Sodium glycochenodeoxycholate (SGCDC), Sodium glycoursodeoxycholate (SGUDC), or a free acid or potassium salt thereof, preferably one or both of Sodium taurodeoxycholate (STDC), Sodium glycodeoxycholate (SGDC); preferably the flavonoid glycoside is in a weight ratio to the surfactant in the range of E-F, wherein E is 1:0.05 and F is 1:9.

1.

9. The supramolecular nanocomplex of claims 1-8, wherein The supramolecular nanocomplex further comprises povidone and / or copovidone and / or polyvinylcaprolactam-polyvinylacetate-polyethylene glycol graft copolymer (Soluplus), preferably the flavonoid glycoside of formula I and povidone and / or copovidone and / or polyvinylcaprolactam-polyvinylacetate-polyethylene glycol graft copolymer (Soluplus) are in a weight ratio in the range of G-H, wherein G is 1 :0.1 and H is 1 :1.

1.

10. A composition, preferably a pharmaceutical composition, comprising the supramolecular nanocomplex according to any one of claims 1 to 9, and an excipient.

11. The supramolecular nanocomplex or composition of any one of claims 1-10, wherein, The supramolecular nanocomplex or composition is in a form suitable for oral administration, after oral administration the active substance is released from the stomach, or from the duodenum, or from the jejunum, or from the ileum, or from the colon or from the rectum.

12. The supramolecular nanocomplex or composition of any one of claims 1-10, wherein, The supramolecular nanocomplex or composition is in a form suitable for rectal administration, preferably delivered to the colon or rectum.

13. The composition according to any one of claims 10 to 12, characterized in that, The composition is in a form for fast release or slow release.

14. The composition according to claim 13, characterized in that, It is in a form suitable for rectal, oral administration, for example, the composition is in the form of a tablet (preferably enteric or colon soluble tablet), microtablet, capsule, pellet, microcapsule, enema, foam, suppository, gel (such as in situ gel), or granule. The supramolecular nanocomplex further comprises povidone and / or copovidone and / or polyvinylcaprolactam-polyvinylacetate-polyethylene glycol graft copolymer (Soluplus), preferably the flavonoid glycoside of formula I and povidone and / or copovidone and / or polyvinylcaprolactam-polyvinylacetate-polyethylene glycol graft copolymer (Soluplus) are in a weight ratio in the range of G-H, wherein G is 1 :0.1 and H is 1 :1.

1.

10. A composition, preferably a pharmaceutical composition, comprising the supramolecular nanocomplex according to any one of claims 1 to 9, and an excipient. The supramolecular nanocomplex or composition is in a form suitable for oral administration, after oral administration the active substance is released from the stomach, or from the duodenum, or from the jejunum, or from the ileum, or from the colon or from the rectum. The supramolecular nanocomplex or composition is in a form suitable for rectal administration, preferably delivered to the colon or rectum. The composition is in a form for fast release or slow release. It is in a form suitable for rectal, oral administration, for example, the composition is in the form of a tablet (preferably enteric or colon soluble tablet), microtablet, capsule, pellet, microcapsule, enema, foam, suppository, gel (such as in situ gel), or granule.

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