CN118845791B - Crocetin preparation with high bioavailability and application thereof - Google Patents
Crocetin preparation with high bioavailability and application thereof Download PDFInfo
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Abstract
本发明属于医药制剂技术领域，提供了一种高生物利用度的西红花酸制剂及其应用。本发明以西红花酸钠为主要活性成分，首次提出将其制备成生物利用度高、吸收快的口服肠溶制剂，通过优化口服制剂处方工艺，以脂质材料、增溶剂和pH调节剂作为西红花酸钠肠溶制剂的关键成分，显著增加了药物成分的稳定性、溶解度及吸收效率，提高了难溶性西红花酸制剂口服的生物利用度，改善了药物的药代动力学参数及疗效。本发明的口服肠溶制剂能有效预防或治疗心脑血管疾病、代谢疾病，极大扩展了西红花酸的临床价值及成药性，且制备工艺简单、成本低。
The present invention belongs to the technical field of pharmaceutical preparations, and provides a crocetin preparation with high bioavailability and its application. The present invention takes sodium crocetin as the main active ingredient, and proposes for the first time to prepare it into an oral enteric-coated preparation with high bioavailability and fast absorption. By optimizing the oral preparation prescription process, lipid materials, solubilizers and pH regulators are used as key components of sodium crocetin enteric-coated preparations, which significantly increase the stability, solubility and absorption efficiency of the drug ingredients, improve the oral bioavailability of poorly soluble crocetin preparations, and improve the pharmacokinetic parameters and efficacy of the drug. The oral enteric-coated preparation of the present invention can effectively prevent or treat cardiovascular and cerebrovascular diseases and metabolic diseases, greatly expand the clinical value and drugability of crocetin, and the preparation process is simple and the cost is low.
Description
Technical Field
The invention belongs to the technical field of medicinal preparations, and in particular relates to a crocetin preparation with high bioavailability and application thereof.
Background
Stigma croci is a dry stigma of croci Crocus sativus L of Iridaceae, has mild taste, and is effective in promoting blood circulation, dispelling blood stasis, cooling blood, removing toxic substances, resolving stagnation, and tranquilizing mind. Crocetin is one of the active ingredients of crocus sativus, also called crocetin, exists in crocus sativus and gardenia plants, is a natural carotenoid dicarboxylic acid, has a molecular formula of C 20H24O4 and a molecular weight of 328.40g/mol. Crocetin is a hydrophobic compound, insoluble in water and most organic solvents, and its anionic form is soluble in water, readily soluble in dilute aqueous sodium hydroxide or other alkaline aqueous solutions. Because crocetin is a polyene compound with conjugated double bonds and taking isoprene as a residue, the crocetin has the advantages of poor structural stability, low bioavailability and easy degradation, and influences the safety and effectiveness of the medicament. Along with the continuous and deep research of the crocetin, the crocetin has various pharmacological effects of protecting heart, protecting nerves, protecting liver, resisting depression, improving Alzheimer disease, parkinsonism and the like, and has great development potential.
In modern pharmacology, crocetin has various curative effects of improving oxygen diffusivity, preventing atherosclerosis, reducing blood fat, reducing blood pressure, increasing eye blood flow and the like, because crocetin has remarkable antioxidant effect. Research shows that the crocetin can effectively remove free radicals in vivo, activate mitogen activated protein kinase (mitogen-ACTIVATED PROTEIN KINASE, MAPK) and PI3K/ATK channels, reduce Reactive Oxygen Species (ROS) production and myocardial apoptosis, and further play a role in heart failure protection. In clinical experiments, crocetin can inhibit mitochondrial function by scavenging free radicals, activate AMPK pathway, reduce intracellular fat oxidation and expression of cell immune antigens at focus parts, promote glucose uptake, accelerate fat consumption and reverse myocardial injury. Lautenschlager et al demonstrate that crocetin can penetrate the blood brain barrier, which is probably the mechanism by which crocetin exerts its efficacy in the central nervous system, making crocetin have the potential to prevent and treat cardiovascular and cerebrovascular diseases.
However, the crocetin related preparation is limited by factors such as high price, poor solubility and stability, extremely low bioavailability and the like of crocetin, and the bioavailability of the medicament is improved by intravenous injection in clinical experiments at present, but the half life of the medicament is very short. The maximum plasma concentration of crocetin was reached about 0.5 hours after administration, indicating that rapid absorption of crocetin may be due to transport into the blood stream via the portal vein. In clinical experiments of healthy human bodies, the concentration of crocetin at different sampling intervals after single oral administration of 16mg of crocetin ranges from about 0.09 to 0.35 mug/mL. Clinical experimental observation of pharmacokinetics of 10 healthy human bodies shows that 7.5, 15 and 22.5mg of crocetin can be detected in human plasma after being taken for 1h by single administration of the crocetin, the t max range is 4.0-8 h, the t 1/2 ranges from 6.1 h to 7.5h, the average values of C max and AUC 0-24h are respectively 100.9-279.7ng/mL and 556.5-1720.8ng/mL, and the crocetin is dose-dependent and still very low compared with crocetin metabolite-crocetin monoglucuronic acid. The oral bioavailability of crocetin is only 11.25% when administered to rats by gavage at a dose of 25 mg/kg. Due to the rapid absorption and metabolism of crocetin, repeated oral administration of crocetin does not accumulate in blood plasma to increase blood concentration, and the bioavailability of crocetin is still low. Therefore, the problem of improving the bioavailability of crocetin is urgently needed to be solved.
At present, in order to improve the bioavailability and stability of crocetin, a plurality of novel drug delivery systems exist, in the study of Zhou et al, gum arabic is used as a wall material to prepare crocetin microcapsules, half life and stability of crocetin are improved, but the defects that the microcapsules cannot be continuously produced, the production process of microcapsule drugs cannot be continuously realized, and the efficiency is low exist. In addition, the drug release rate of the microencapsulated drug may have an unstable problem. Langroodi et al adopts a W/O/W multiple emulsion method to prepare crocetin PLGA nanoparticles, so that the slow release effect and pharmacological activity of crocetin are improved, but the preparation has the problems of high production cost, preparation complexity and the like. El-Kharrag et al prepared magnetic nanoparticles by co-precipitation with FeCl 2·4H2O、FeCl3 and dextran-containing NaOH solution, and then performing in-situ inclusion. Research shows that the dispersibility and stability of the crocetin in water are improved, but the defect is that the effect of the magnetic nanoparticle medicament is influenced by the strength and the direction of a magnetic field, the production and use cost of the magnetic nanoparticle medicament is relatively high, and the wide clinical application of the magnetic nanoparticle medicament can be limited. Esposito et al uses glycerol monooleate, sodium cholate and sodium caseinate as raw materials to prepare a crocetin nanostructured lipid dispersion, so that the stability and pharmacological activity of crocetin are improved, but the problems of high production cost and the like are faced, and the storage condition of the nanostructured lipid dispersion is more severe.
Chinese patent CN109091458a discloses a method for preparing crocetin microemulsion and freeze-dried powder thereof, based on the principle of microemulsion, the surfactant added in the preparation process increases the solubility of crocetin in water by 800 times, but also affects the stability while improving the solubility. In addition, chinese patent CN112999162a discloses a solid dispersion of crocetin and a preparation method thereof, wherein hydrophilic polymer carrier material and meglumine are dissolved and then mixed, crosslinked with each other to form carrier polymer, and then mixed and dissolved with crocetin, and the solid dispersion is prepared by solvent evaporation method or spray drying method. The foreign patent WO2022/025997A1 discloses a trans-crocetin composition and a treatment scheme, wherein the provided trans-crocetin medicament and the provided administration scheme remarkably increase the solubility of crocetin and improve the stability of crocetin, but the preparation process of a trans-liposome preparation is complex and the production cost is high.
The invention aims to research a preparation which can be produced in a technological way and has low cost. The sodium salt is prepared from crocetin, and then the prescription and the process of the sodium crocetin oral preparation are optimized, so that the problem that crocetin is insoluble in water and sodium crocetin is extremely unstable in dissolution is solved. Finally, the crocetin preparation with high bioavailability and the application thereof are provided, the half life of the medicament is effectively prolonged, the solubility and the solution stability of Gao Xigong florin are improved, the bioavailability of the crocetin preparation is improved, and the clinical curative effects of treating cardiovascular and cerebrovascular diseases, nervous system diseases and the like are further improved. The preparation process has strong operability and low cost, is suitable for industrial expansion production, and has good clinical application prospect.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a crocetin preparation with high bioavailability and application thereof. Specifically, the enteric-coated preparation is prepared by taking sodium crocetin as a main active ingredient, so that the stability and the solubility of the medicine are obviously improved, and the metabolism condition of sodium crocetin in vivo is effectively improved, thereby improving the bioavailability and the absorptivity of the medicine, effectively preventing or treating cardiovascular and cerebrovascular diseases, and greatly expanding the drug property and the clinical value of crocetin.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
In one aspect, the invention provides a crocetin preparation with high bioavailability, which is prepared by preparing crocetin active ingredients into an oral enteric preparation, wherein the crocetin active ingredients are at least one of crocetin and sodium crocetin.
Preferably, the formulation comprises at least one of a pH adjuster, a solubiliser, a lipid material in addition to the saffron active ingredient.
Further preferably, the formulation comprises safflower active ingredient, pH adjuster, solubilizing agent and lipid material.
Still more preferably, the pH adjuster is at least one selected from sodium bicarbonate, sodium alginate, meglumine, sodium hydroxide, and disodium hydrogen phosphate.
Still more preferably, the solubilizing agent is at least one selected from hydroxypropyl methylcellulose, hydroxypropyl beta cyclodextrin, povidone, copovidone, hydroxypropyl cellulose, polyethylene glycol-15 hydroxystearate, vitamin E polyethylene glycol succinate.
Still more preferably, the lipid material is selected from at least one of glyceryl monostearate, glyceryl behenate, hydrogenated castor oil.
Still more preferably, the pH adjuster is sodium bicarbonate.
Still more preferably, the solubilizing agent is at least one selected from hydroxypropyl methylcellulose, vitamin E polyethylene glycol succinate.
Still more preferably, the lipid material is glyceryl behenate.
Still more preferably, the crocetin preparation comprises 1 part by weight of crocus sativus active ingredient, 1-10 parts by weight of a pH regulator, 1-20 parts by weight of a solubilizing agent and 1-20 parts by weight of a lipid material.
Still more preferably, the crocetin preparation comprises 1 part by weight of crocus sativus active ingredient, 1-8 parts by weight of pH regulator, 1-15 parts by weight of solubilizer and 1-15 parts by weight of lipid material.
Still more preferably, the crocetin preparation comprises 1 part by weight of crocus sativus active ingredient, 1-5 parts by weight of pH regulator, 1-10 parts by weight of solubilizer and 1-10 parts by weight of lipid material.
Preferably, the crocetin preparation further comprises a filler, a glidant, a lubricant and an enteric auxiliary material.
Further preferably, the filler is at least one selected from lactose, soluble starch, microcrystalline cellulose, dextrin, dibasic calcium phosphate, mannitol, pregelatinized starch.
Further preferably, the glidant is at least one selected from the group consisting of colloidal silicon dioxide and talc.
Further preferably, the lubricant is at least one selected from magnesium stearate, calcium stearate, micro silica gel, hydrogenated vegetable oil, polyethylene glycol, and sodium dodecyl sulfate.
Further preferably, the weight gain of the enteric auxiliary material after coating is 8% -20%.
Further preferably, the enteric adjuvants may be commercially available products, and can be routinely selected by those skilled in the art to ensure that the formulation does not dissolve in stomach acid but rather dissolves in the small intestine.
Preferably, the formulation is selected from any one of enteric-coated tablets, enteric-coated capsules, enteric-coated granules and enteric-coated pellets.
In still another aspect, the present invention also provides a preparation method of the crocetin preparation, comprising the following steps:
S1, preprocessing, namely mixing the saffron active ingredient and the filler to form a mixture 1, and sieving the mixture 1;
S2, granulating, namely mixing the mixture 1, the slow release agent and the solubilizer to form a mixture 2, dissolving the stabilizer and the other solubilizer in water to form a solution, mixing the mixture 2 and the solution, performing wet granulation to form wet granules, and sieving the wet granules;
s3, drying and granulating, namely drying and sieving wet granules to obtain dry granules;
S4, mixing the glidant, the lubricant and the dry particles to form a total mixture;
s5, tabletting, namely tabletting the total mixture to form a plain tablet;
s6, curing and coating, namely curing, isolating and coating and enteric coating the plain tablets in sequence to obtain the crocetin preparation.
Preferably, in step S1, the mesh number of the sieving is 80 mesh.
Preferably, in step S2, the mesh number of the sieving is 40 mesh.
Preferably, in step S3, the moisture content of the wet granules after drying is less than 2.5%.
Preferably, in step S3, the mesh number of the sieving is 30 mesh.
Preferably, in step S5, the tabletting is performed using a rotary tablet press.
Preferably, in step S5, the hardness of the plain film is 70±20N.
Preferably, in the step S6, the specific condition of curing is that the temperature is 60-80 ℃ and the time is 1-5h.
In still another aspect, the invention also provides application of the crocetin preparation in preparing medicaments for treating cardiovascular and cerebrovascular diseases.
Preferably, the cardiovascular and cerebrovascular diseases comprise coronary heart disease, angina pectoris, heart failure, myocardial infarction, hyperlipidemia and metabolic syndrome.
Compared with the prior art, the invention solves the technical problem of poor crocetin patent medicine, obtains unexpected technical effects, obviously improves bioavailability and pharmacological activity in vivo of oral administration, and has the following obvious beneficial effects:
(1) According to the invention, the enteric preparation is prepared by taking the crocetin as a main active ingredient for the first time, the lipid material, the solubilizer and the pH regulator are taken as key ingredients of the crocetin enteric preparation by optimizing a prescription process, and the lipid material and the solubilizer can both improve the dissolution rate of the crocetin, and under the synergistic effect of the lipid material, the solubilizer and the pH regulator, the dissolution rate of the crocetin can be obviously improved, and the solution stability can also be improved. The prescription process solves the key technical problems of the patentability such as the insolubility of the crocetin in water, poor solution stability and the like.
(2) The invention discovers for the first time that the sodium crocetin is prepared into a sample with high water solubility, and the bioavailability of crocetin can be remarkably improved by the administration of duodenum, which is 9.56 times of that of an oral gastric lavage administration group. Further, the bioavailability of the enteric preparation provided by the invention can be improved to more than 30 times of that of oral crocetin, and the enteric preparation is quick to absorb and long in half-life.
(3) The pH regulator, the lipid material and the solubilizer in the enteric preparation prepared by the invention can synergistically promote the dissolution of the crocetin, and the accelerated experiments prove that the dissolution phenomenon is not seen under the condition of six months of acceleration, so that the stability of the crocetin is obviously improved. The drug effect comparison research of the myocardial infarction rat heart failure model is carried out by adopting the conventional enteric preparation and the full-prescription enteric preparation, and the drug substitution and the drug effect of the full-prescription enteric preparation are found to be obviously superior to those of crocetin and are also obviously superior to those of the conventional enteric preparation prescription sample. The full-prescription enteric preparation of the invention can obviously improve the disease development process of heart failure diseases and various indexes of heart, can effectively treat or prevent cardiovascular and cerebrovascular diseases, and greatly expands the clinical value and the drug property of crocetin.
(4) Through extensive literature investigation and analysis, the invention combines years of research experience and continuous experimental exploration of the inventor, finally successfully solves the problems of water-insoluble crocetin, poor solution stability and other patent medicines, breaks through the technical difficulties of the existing nano preparation, liposome and other complex preparations, breaks through the limitation that the crocetin preparation can only be injected and administered, and reduces the cost. The invention discloses a brand new crocetin enteric preparation which is suitable for industrial production and low in cost, and the bioavailability, the treatment effect and the clinical application value of crocetin are obviously improved.
Drawings
FIG. 1 is a graph showing the results of an experiment simulating the cumulative dissolution rate of intestinal juice.
FIG. 2 shows the effect of each group of doses on NT-proBNP in rats in a comparative pharmacodynamics experiment of the enteric crocetin preparation.
FIG. 3 shows the effect of each group of doses on cardiac index of rats in a pharmacodynamic comparison experiment of enteric crocetin formulation.
Detailed Description
The following non-limiting examples will enable those of ordinary skill in the art to more fully understand the application and are not intended to limit the application in any way. The following is merely exemplary of the scope of the application as claimed and many variations and modifications of the application will be apparent to those skilled in the art in light of the disclosure, which are intended to be within the scope of the application as claimed.
The invention is further illustrated by means of the following specific examples. The various chemical reagents used in the examples of the present invention were obtained by conventional commercial means unless otherwise specified. Unless otherwise specified, the contents are mass contents in the following. Unless otherwise indicated, it is understood that it is carried out at room temperature.
Crocetin (CRA) and sodium Crocetin (CRAT) are provided by the preparation of apricot forest chinese medicine technologies (guangzhou) limited.
Example 1 (comparative experiments on pharmacokinetics of sodium saffron different routes of administration)
1. Subject configuration
Preparing a CRAT intravenous administration test sample, namely preparing administration liquid medicine with the concentration of 1.5mg/mL of CRAT by using 5% glucose, and filtering and sterilizing before administration;
the preparation of test products for gastric administration and intestinal intubation of CRAT comprises diluting with 0.5% sodium carboxymethylcellulose (CMC-Na) to prepare administration suspension with CRAT concentration of 1.5 mg/mL;
The preparation of test sample for gastric administration of CRA comprises diluting with CMC-Na of 0.5% to obtain administration suspension with CRA concentration of 1.5 mg/mL.
2. Experimental animal
24 SD rats, SPF grade, weight 223.3-244.9g, age of 6-7 weeks, male and female halves. Is provided by Hunan Laike Jingda laboratory animal Limited, laboratory animal quality certificate number No430727201101012371, laboratory animal production license number SCXK (Hunan) 2019-0004. The raising condition is that the temperature is 20-26 ℃, the humidity is 40-70%, the day and night light and shade alternation time is 12h/12h, and the rats drink water freely. After one week of adaptive feeding, the rats were subjected to experiments.
3. Grouping and administration
The quarantined SFP-grade SD rats were randomly divided into 4 groups according to sex weight, each group of which has 3 male and female animals, and sodium crocetin intravenous administration group (IV), crocetin intragastric administration group (CRA), sodium crocetin intragastric administration group (CRAT), sodium crocetin enteric administration group (DI) (from duodenum) were used for comparative study. The experimental animals were fasted for 12 hours before administration, were free to drink water, were given the corresponding subjects according to the dosing schedule of table 1 the following day, were fasted for 2 hours after administration, and were fasted for 4 hours.
TABLE 1 grouping of subjects and dosing regimen
| Dose group | Group of | Dosage/mg/kg | Test article | Administration mode | Number of animals |
| Intravenous administration group | IV | 5 | CRAT | Intravenous injection | 3 Male and female animals |
| Gastric lavage administration group | CRA | 15 | CRA | Stomach lavage | 3 Male and female animals |
| Gastric lavage administration group | CRAT | 15 | CRAT | Stomach lavage | 3 Male and female animals |
| Intestinal intubation administration group | DI | 15 | CRAT | Duodenal administration | 3 Male and female animals |
The above dosage is the dosage of crocetin.
4. Sample collection
Whole blood was collected at the corresponding time points from about 0.25mL to about 0.25mL of pre-labeled heparinized blood collection tubes in the orbital venous plexus (isoflurane inhalation anesthesia) or jugular vein, wherein blood samples were collected for each group of animals 5min, 10min, 20min, 45min, 1.5h, 3h, 6h, 12h, 24h before and after dosing, respectively. The blood sample is put in an ice box for temporary storage, then centrifuged for 10min at 4500rpm, the plasma is separated, the separated plasma is put in a marked centrifuge tube, and put in an ultra-low temperature refrigerator for temporary storage and test.
5. Detecting blood concentration of each group by high performance liquid chromatograph
The pharmacokinetic parameters of each group of rats were counted and the main pharmacokinetic parameters of CRA in plasma after each administration group are shown in Table 2.
Table 2. Summary of essential pharmacokinetic parameters for each dosing group (n=6)
As can be seen from the results in Table 2, the AUC (0-t) of the CRAT enteral administration group is 77305.3 + -23733.1 μg/L h, the AUC (0-t) of the CRAT intragastric administration group is 8086.2 + -1872.5 μg/L h, the bioavailability of the enteral administration group is 9.56 times that of the intragastric administration group, 32.86 times that of the isodose CRA intragastric administration group, the intravenous administration t 1/2 is 0.67+ -0.17 h, the intragastric administration t 1/2 is 9.64+ -13.09 h, the duodenal administration t 1/2 is 4.65+ -2.62 h, suggesting rapid metabolism after CRA blood infusion, short drug half-life and rapid peak arrival time, resulting in C max of the intravenous administration group being higher than that of the enteral administration group.
In addition, the water solubility of crocetin is improved after salification, the bioavailability is improved by about 4 times compared with that of crocetin, but the absolute bioavailability of CRA and CRAT gastric administration groups is respectively 1.62 percent and 5.58 percent, both are extremely low, and the absolute bioavailability of CRAT duodenal administration group is 53.39 percent which is obviously higher than that of the gastric administration group. It can be seen that although CRAT has improved water solubility, conventional gastric-soluble formulations still suffer from insufficient drug-forming properties. According to comprehensive analysis, sodium crocetin is more suitable for being clinically made into an enteric preparation, so that the invention further researches the enteric preparation.
Example 2 (examination of the pH adjustor by crocetin enteric preparation)
Through long-time preparation research and dissolution investigation, the sodium saffron or the crocetin is prepared into samples by adopting a conventional prescription process, and the dissolution rate of the qualified preparation meeting the dissolution rate requirement of the pharmacopoeia preparation is not achieved, and the dissolution rate of the qualified preparation in a conventional neutral or slightly acidic medium is extremely low. Crocetin has extremely low solubility in neutral and acidic environments and cannot be dissolved out. In the neutral and slightly acidic environment, the particles show that sodium crocetin is extremely easy to hydrolyze into crocetin, so that sodium crocetin and crocetin samples cannot be dissolved out smoothly in a conventional medium. In view of this, in addition to reducing the particle size of the drug substance and increasing the dispersion of the drug substance to the greatest extent, an alkaline adjuvant is introduced into the formulation for increasing the pH of the drug substance particle microenvironment, thereby increasing the dissolution of the drug substance, and the effect of the pH adjuster on the enteric preparation is examined first, and the formulation is shown in table 3.
TABLE 3 formulation of enteric crocetin formulations
The "/" indicates that no addition was made, as follows.
The preparation process of prescriptions 1-5 is as follows:
s1, preprocessing, namely crushing crocetin and lactose for 30 seconds by a high-shear crusher, and sieving the crushed mixture with a 80-mesh sieve;
S2, granulating, namely placing the mixture, microcrystalline cellulose and crospovidone into a 5L wet granulating pot, stirring at 300rpm, cutting at 2000rpm, and mixing for 5min. Adding pH regulator into purified water for dissolution, pouring the solution into a wet granulating pot for wet granulating, stirring at 300rpm and cutting at 2000rpm, granulating for 3min, and sieving the obtained soft material with a 40 mesh sieve;
s3, drying and granulating, namely placing wet granules in a blast drying oven at 60 ℃ for drying for 2 hours, controlling the moisture of the granules to be within 2.5%, and granulating the obtained dry granules by using a 30-mesh sieve;
s4, total mixing, namely adding colloidal silicon dioxide, magnesium stearate and the dry particles into a square conical mixer barrel 5L, and mixing for 5min to obtain a total mixture.
S5, tabletting, namely tabletting the total mixture by adopting a rotary tablet press, wherein the hardness is controlled to be 100+/-20N, and thus the plain tablets are obtained.
The in vitro dissolution rates (high performance liquid chromatography) of prescriptions 1-5 were determined and the detection results were as follows;
The in vitro dissolution method comprises taking phosphate buffer solution with pH of 6.0 as dissolution medium, measuring volume of the dissolution medium at 900mL, rotating at 50 r/min, performing dissolution experiment by slurry method, sampling and supplementing 10mL each time, measuring dissolution result by high performance liquid chromatography, and calculating cumulative dissolution rate, wherein the result is shown in Table 4.
TABLE 4 cumulative dissolution rate
| Time/min | 30 | 60 | 120 | 240 |
| Prescription 1 | 2.13% | 2.21% | 2.30% | 2.41% |
| Prescription 2 | 5.44% | 5.49% | 5.55% | 5.62% |
| Prescription 3 | 7.14% | 7.40% | 7.54% | 7.49% |
| Prescription 4 | 2.31% | 2.28% | 2.30% | 2.21% |
| Prescription 5 | 2.82% | 2.81% | 2.84% | 2.83% |
As can be seen from comparison of prescriptions 1-5, crocetin is basically not dissolved in a medium with pH of 6.0, the surface of the raw material medicine particles is easy to acidify and hydrolyze to crocetin, so that the raw material medicine is basically not dissolved, the final dissolution rate is only about 2%, and after common alkaline pH regulator auxiliary materials (meglumine, sodium bicarbonate, sodium alginate and sodium hydroxide) are added into the prescriptions, the solubility is improved, but the degree of improvement is greatly different. The results show that when sodium bicarbonate is used as an alkaline auxiliary material, the solubilization effect is most obvious, the meglumine effect is slightly poor, and sodium hydroxide and sodium alginate have little effect.
Example 3 (examination of the solubilizer by the enteric-coated crocetin preparation)
The amount of dissolution change in the intestinal pH environment is limited only by the addition of pH modifiers. The dissolution rate of sodium crocetin needs to be further improved, and 10 prescriptions are planned to be studied by considering that crocetin has long hydrophobic conjugated double bonds and two hydrophilic terminal carboxyl groups and has the property similar to a surfactant, so the invention considers that a solubilizer is introduced into a prescription, screens different solubilizers, and compares available solubilizers by analyzing the characteristics and the result of a future experiment of the composition, and particularly shown in table 5.
TABLE 5 prescription of enteric crocetin formulation
The preparation process of the prescription 6-11 is as follows:
s1, preprocessing, namely crushing sodium saffron and lactose for 30 seconds by a high-shear crusher, and sieving the crushed mixture with a 80-mesh sieve;
S2, granulating, namely placing the mixture, microcrystalline cellulose, crosslinked povidone and solubilizer (hydroxypropyl methylcellulose, hydroxypropyl beta cyclodextrin, povidone VA064, copovidone, hydroxypropyl cellulose and polyethylene glycol 6000) into a 5L wet granulating pot, stirring at 300rpm and cutting at 2000rpm, and mixing for 5min. Pouring purified water into a wet granulating pot for wet granulating, wherein the stirring speed is 300rpm, the cutting speed is 2000rpm, granulating is carried out for 3min, and the obtained soft material is sieved by a 40-mesh sieve;
s3, drying and granulating, namely placing wet granules in a blast drying oven at 60 ℃ for drying for 2 hours, controlling the moisture of the granules to be within 2.5%, and granulating the obtained dry granules by using a 30-mesh sieve;
s4, total mixing, namely adding colloidal silicon dioxide, magnesium stearate and the dry particles into a square conical mixer barrel 5L, and mixing for 5min to obtain a total mixture.
S5, tabletting, namely tabletting the total mixture by adopting a rotary tablet press, wherein the hardness is controlled to be 100+/-20N, and thus the plain tablets are obtained.
The preparation process of the prescription 12-15 is consistent with the prescription 6-11 except the granulating process, and the granulating process is as follows:
The granulation parameters used remained the same as in formulas 6-11, but the solubilizers (sodium dodecyl sulfate, tween 80, polyethylene glycol-15 hydroxystearate, vitamin E polyethylene glycol succinate) were first dissolved in purified water separately and wet granulated.
The in vitro dissolution rate of the prescription 6-15 is measured, and the detection result is as follows;
The in vitro dissolution method comprises taking phosphate buffer solution with pH of 6.0 as dissolution medium, measuring volume of the dissolution medium at 900mL, rotating at 50 r/min, performing dissolution experiment by slurry method, sampling and supplementing 10mL each time, measuring dissolution result by high performance liquid chromatography, and calculating cumulative dissolution rate, wherein the result is shown in Table 6.
TABLE 6 cumulative dissolution rate
| Time/min | 30 | 60 | 120 | 240 |
| Prescription 6 | 11.88% | 11.07% | 10.21% | 8.75% |
| Prescription 7 | 32.12% | 11.74% | 8.47% | 3.45% |
| Prescription 8 | 9.76% | 8.84% | 7.49% | 7.27% |
| Prescription 9 | 8.44% | 6.42% | 5.55% | 4.62% |
| Prescription 10 | 7.28% | 7.02% | 6.98% | 6.75% |
| Prescription 11 | 6.24% | 6.04% | 5.89% | 5.92% |
| Prescription 12 | 4.42% | 1.45% | 1.87% | 1.54% |
| Prescription 13 | 4.25% | 1.57% | 1.38% | 1.70% |
| Prescription 14 | 18.45% | 18.14% | 16.74% | 15.58% |
| Prescription 15 | 24.41% | 23.89% | 23.14% | 22.12% |
Compared with the prescription 6-15, the hydroxypropyl methylcellulose, the hydroxypropyl beta cyclodextrin, the polyethylene glycol-15 hydroxystearate and the vitamin E polyethylene glycol succinate are used as solubilizers to have a more remarkable solubilization effect on sodium saffron, and other auxiliary materials have a slightly solubilization effect, so that the sodium dodecyl sulfate and the Tween 80 cannot be compatibilized, but can accelerate the precipitation of the raw materials. The hydroxypropyl beta cyclodextrin can obviously improve the dissolution rate of the raw material medicine in a short time, the concentration of the hydroxypropyl beta cyclodextrin is too high to promote the raw material medicine to be subjected to severe hydrolysis, so that the concentration of the final dissolved solution is lower, and then the concentration of the raw material medicine is not obviously reduced in the dissolution process of a sample prepared from vitamin E polyethylene glycol succinate and polyethylene glycol-15 hydroxystearate, and the two can form dark red emulsion drops with the dissolved crocetin, so that crocetin micelle is presumably formed. The macromolecular compound has a certain solubilization effect because most of the macromolecules have a plurality of hydrophilic hydroxyl groups, and can reduce the activity of the raw material medicine molecules, wherein the hydroxypropyl methylcellulose is the most obvious.
Example 4 (examination of lipid materials by enteric crocetin formulations)
The dissolution rate of sodium crocetin in the meta-acidic medium can be improved by adding a pH regulator or a solubilizer in the prescription of the preparation, but the dissolution rate is still lower, and the cumulative dissolution rate is not more than 30%. The sample still fails to realize the cumulative dissolution rate reaching more than 50% through the preparation process and prescription adjustment. The dissolution results of all samples in comparative examples 2 and 3 show that the samples reach the dissolution end point within 30min and are partially hydrolyzed, which is unfavorable for the absorption of the drug in vivo, so the invention finds that the lipid material can provide a tiny space for the dissolution of sodium crocetin due to the insolubility in the dissolution medium through exploratory studies, and considers whether the characteristic can be used or not to improve the dissolution rate and the solution stability of sodium crocetin. Thus, in order to further improve the dissolution rate and solution stability of crocetin, each lipid material characteristic was analyzed, and lipid materials which could achieve the object were finally selected by pre-test, and re-study of the prescription process was performed, and specific designs are shown in table 7.
TABLE 7 formulation of enteric crocetin formulations
The preparation process of prescriptions 16-23 is as follows:
s1, preprocessing, namely crushing sodium saffron and lactose for 30 seconds by a high-shear crusher, and sieving the crushed mixture with a 80-mesh sieve;
S2, granulating, namely placing the mixture, lipid materials (glyceryl monostearate, glyceryl behenate and hydrogenated castor oil) and hydroxypropyl methylcellulose into a 5L wet granulating pot, wherein the stirring speed is 300rpm, the cutting speed is 2000rpm, and mixing for 5min. Dissolving sodium bicarbonate and vitamin E polyethylene glycol succinate in purified water, pouring the solution into a wet granulation pot for wet granulation, stirring at 300rpm, cutting at 2000rpm, granulating for 3min, and sieving the obtained soft material with 40 mesh sieve;
s3, drying and granulating, namely placing wet granules in a blast drying oven at 60 ℃ for drying for 2 hours, controlling the moisture of the granules to be within 2.5%, and granulating the obtained dry granules by using a 30-mesh sieve;
s4, total mixing, namely adding colloidal silicon dioxide, magnesium stearate and the dry particles into a square conical mixer barrel 5L, and mixing for 5min to obtain a total mixture.
S5, tabletting, namely tabletting the total mixture by adopting a rotary tablet press, wherein the hardness is controlled to be 70+/-20N, and thus the plain tablets are obtained.
The in vitro dissolution rates and contents of the formulations 16-23 were determined (high performance liquid chromatography) and the detection results were as follows;
The in vitro dissolution method comprises taking phosphate buffer solution with pH of 6.0 as dissolution medium, measuring volume of the dissolution medium at 900mL, rotating at 50 r/min, performing dissolution experiment by slurry method, sampling and supplementing 10mL each time, measuring dissolution result by high performance liquid chromatography, and calculating cumulative dissolution rate, wherein the result is shown in Table 8.
TABLE 8 cumulative dissolution rate
As can be seen from the results of the dissolution test in Table 8, comparison of the formulations 18, 20, and 22 with the formulation 16 shows that the lipid material can improve the dissolution of crocetin, but the problem of low dissolution cannot be solved by adding only the lipid material. From the previous examples, the addition of only both pH adjustor and solubilizer did not meet the dissolution requirements. The dissolution results of the prescriptions 19, 21 and 23 meet the pharmacopoeia requirements, and are obviously higher than those of all other prescription process samples, which shows that the lipid materials in the prescriptions have synergistic effect together with the pH regulator and the solubilizer, and the solubility and the solution stability of sodium crocetin can be obviously improved. Whether the bioavailability and the drug effect can be obviously improved or not, and the further study is needed.
In addition, in view of the fact that glyceryl behenate can serve as a fat-soluble material to provide a stable microenvironment for dissolution of bulk drugs, the fat-soluble material is unevenly distributed in a freshly compressed tablet, so that the formed microenvironment is uneven or unstable, and in view of the fact that the melting point of the glyceryl behenate is 65-70 ℃, the tablet is cured for 2 hours under the condition of 70 ℃ so that the distribution of the tablet is more even, and the formed microenvironment is smaller, more uniform and more stable. The results of in vitro elution (elution method is the same as above) of the cured sample prepared in the formulation 21 are shown in Table 9.
TABLE 9 cumulative dissolution rate of samples prepared in prescription 21 after maturation
| Time/min | 30 | 60 | 120 | 240 | Content of finished product |
| Prescription 21-after curing | 21.41% | 48.79% | 84.49% | 99.73% | 100.31% |
Example 5
Conventional enteric coated tablets were prepared as in example 2, formulation 1, and the enteric coated tablet formulation did not contain pH adjuster, solubilizer, lipid material.
The tablet of the prescription 1 of the example 2 is firstly subjected to isolation coating by using the Ophiopogon 88A180040-CN, the weight is increased by about 3 percent, then the tablet is subjected to enteric coating by using the Ophiopogon 93O65142-CN, and the weight is increased by about 10 percent, thus obtaining the enteric-coated tablet.
Example 6
An enteric coated tablet (containing a pH adjuster) was prepared as in example 2, formulation 3, which did not contain a solubilizing agent, lipid material.
The tablet of the prescription 3 of the example 2 is firstly subjected to isolation coating by using the Ophiopogon 88A180040-CN, the weight is increased by about 3 percent, then the tablet is subjected to enteric coating by using the Ophiopogon 93O65142-CN, and the weight is increased by about 10 percent, thus obtaining the enteric-coated tablet.
Example 7
An enteric coated tablet (containing pH adjuster and solubilizer) was prepared as in example 4, formulation 15, which did not contain lipid material.
The tablet of the prescription 15 of the example 4 is firstly subjected to isolation coating by using the Ophiopogon 88A180040-CN, the weight is increased by about 3 percent, then the tablet is subjected to enteric coating by using the Ophiopogon 93O65142-CN, and the weight is increased by about 10 percent, thus obtaining the enteric-coated tablet.
Example 8
An enteric coated tablet (whole formulation) containing a pH adjuster, a solubilizing agent, and a lipid material was prepared as in example 4 formulation 21.
The tablet of the prescription 21 of the example 4 is cured for 2 hours at 70 ℃, the cured tablet is firstly subjected to isolation coating by using the Ophio 88A180040-CN, the weight gain is about 3 percent, then the tablet is subjected to enteric coating by using the Ophio 93O65142-CN, and the weight gain is about 10 percent, thus obtaining the enteric tablet.
Example 9
An enteric-coated tablet of crocetin containing a pH adjustor, a solubilizer and a lipid material was prepared as in the formulation (whole formulation) of the formulation 21 of example 4, substituting crocetin for sodium crocetin as a raw material.
S1, preprocessing, namely crushing sodium saffron and lactose for 30 seconds by a high-shear crusher, and sieving the crushed mixture with a 80-mesh sieve;
s2, granulating, namely placing the mixture, the glyceryl behenate and the hydroxypropyl methylcellulose into a 5L wet granulating pot, stirring at 300rpm, cutting at 2000rpm, and mixing for 5min. Dissolving vitamin E polyethylene glycol succinate in purified water, pouring the solution into a wet granulating pot for wet granulating, stirring at 300rpm, cutting at 2000rpm, granulating for 3min, and sieving with 40 mesh sieve;
s3, drying and granulating, namely placing wet granules in a blast drying oven at 60 ℃ for drying for 2 hours, controlling the moisture of the granules to be within 2.5%, and granulating the obtained dry granules by using a 30-mesh sieve;
s4, total mixing, namely adding colloidal silicon dioxide, magnesium stearate and the dry particles into a square conical mixer barrel 5L, and mixing for 5min to obtain a total mixture.
S5, tabletting, namely tabletting the total mixture by adopting a rotary tablet press, wherein the hardness is controlled to be 70+/-20N, and thus the plain tablets are obtained.
S6, baking the slices, namely placing the slices at 70 ℃ and drying for 2 hours to obtain cured slices.
S7, the cured plain tablet is subjected to isolation coating by using the Ophiopogon 88A180040-CN, the weight is increased by about 3%, then the cured plain tablet is subjected to enteric coating by using the Ophiopogon 93O65142-CN, and the weight is increased by about 10%.
Example 10
Enteric pellets were prepared according to example 4, prescription 21 (all-round).
S1, preprocessing, namely crushing sodium saffron and lactose for 30 seconds by a high-shear crusher, and sieving the crushed mixture with a 80-mesh sieve;
S2, granulating, namely placing the mixture, the glyceryl behenate and the hydroxypropyl methylcellulose into a 5L wet granulating pot, stirring at 300rpm, cutting at 2000rpm, and mixing for 5min. Pouring sodium bicarbonate and vitamin E polyethylene glycol succinate solution into a wet granulating pan for wet granulating at stirring speed of 300rpm and cutting speed of 2000rpm, granulating for 3min to obtain soft material;
S3, pelleting, namely pouring the prepared soft material into an extrusion spheronizer, installing an extrusion screen with the thickness of 0.8mm, and starting the extrusion spheronizer to prepare pellets;
S4, drying and finishing, namely transferring the pellets into a fluidized bed, drying at the air inlet temperature of 60 ℃, putting the pellets into a 10-mesh screen for finishing after the materials are dried, and removing fine powder by a 40-mesh screen to obtain dry pellets;
S5, coating, namely transferring the pellets into a fluidized bed, spraying the Ophio 88A180040-CN for isolation coating, increasing the weight by about 3%, and then using the Ophio 93O65142-CN for enteric coating, wherein the weight is increased by about 10%.
Example 11
Enteric coated granules were prepared according to example 4, prescription 21 (all-round).
S1, preprocessing, namely crushing sodium saffron and lactose for 30 seconds by a high-shear crusher, and sieving the crushed mixture with a 80-mesh sieve;
s2, placing the mixture, the glyceryl behenate and the hydroxypropyl methylcellulose into a 5L wet granulation pot, stirring at 300rpm, cutting at 2000rpm, and mixing for 5min. Pouring sodium bicarbonate and vitamin E polyethylene glycol succinate solution into a wet granulating pan for wet granulating at stirring speed of 300rpm and cutting speed of 2000rpm, granulating for 3min to obtain soft material;
s3, granulating, namely pouring the prepared soft material into a rotary granulator, and granulating by adopting a sieve with the aperture of 0.8mm and a rotating speed of 20 rpm;
S4, drying and finishing, namely transferring the particles into a fluidized bed, drying at the air inlet temperature of 60 ℃, putting the dried materials into a 10-mesh screen for finishing after the materials are dried, and removing fine powder by a 40-mesh screen to obtain dry particles;
S5, coating, namely transferring the particles into a fluidized bed, spraying the Ophio 88A180040-CN for isolation coating, increasing the weight by about 3%, and then using the Ophio 93O65142-CN for enteric coating, wherein the weight is increased by about 10%.
Results and discussion
1. Experimental for simulating accumulated dissolution rate of intestinal juice
The in vitro dissolution was measured according to the method 1 for measuring dissolution and release of enteric preparation of the pharmacopoeia 2020 edition of the people's republic of China, 750mL of hydrochloric acid solution of 0.1mol/L was used as a dissolution medium, the rotation speed was 50 revolutions per minute, 0.2mol/L of sodium phosphate solution of 250mL preheated to 37 ℃ plus or minus 0.5 ℃ was immediately added after 120 minutes, the rotation speed was 50 revolutions per minute, the sample was supplemented by 10mL each time of sampling, the dissolution result was measured by high performance liquid chromatography, the formal sampling was started from the time before the addition of sodium phosphate solution, and the cumulative dissolution rate was calculated, and the results are shown in Table 10.
TABLE 10 cumulative dissolution rate
| Time/min | 0 | 30 | 60 | 120 | 240 | 480 |
| Prescription 1 | 0.23% | 1.54% | 1.56% | 1.62% | 1.66% | 1.67% |
| Prescription 21 | 0.33% | 4.34% | 4.25% | 4.75% | 4.74% | 4.77% |
| Example 5 | 0.00% | 3.12% | 3.15% | 3.11% | 3.08% | 3.10% |
| Example 6 | 0.00% | 7.98% | 8.52% | 8.48% | 8.45% | 8.58% |
| Example 7 | 0.00% | 43.47% | 44.51% | 42.12% | 41.08% | 40.72% |
| Example 8 | 0.00% | 30.66% | 59.57% | 92.24% | 99.54% | 99.78% |
| Example 9 | 0.00% | 23.76% | 42.82% | 65.62% | 74.24% | 75.21% |
| Example 10 | 0.00% | 38.43% | 65.48% | 99.48% | 100.20% | 100.42% |
| Example 11 | 0.00% | 44.25% | 78.41% | 98.45% | 98.47% | 98.50% |
The above experimental results show that the preparation of the sample recipe processes of recipe 1, recipe 21 and examples 5-7 do not add the key ingredient combination of the present invention at the same time, and the dissolution rate is too low. The enteric-coated tablet, the enteric-coated pellet and the enteric-coated granule of the crocetin of the examples 8-11 are tested by acid resistance and dissolution, and all reach the aim of achieving the slow release that crocetin can be dissolved rapidly after reaching intestinal tracts, so that the prepared enteric-coated preparation achieves the expected effect of releasing crocetin, namely, the crocetin is completely kept in a specified acid medium (gastric acid) for 2 hours, the medicine is not released or is hardly released, and the dissolution in intestinal juice can almost reach 100 percent, thereby achieving the aim of achieving the slow release that the medicine can not be released in stomach but can be dissolved rapidly after reaching intestinal tracts, and remarkably improving the bioavailability of crocetin for oral administration and the concentration of crocetin in intestinal tracts.
2. Accelerated stability test
The whole enteric-coated tablets prepared in example 8 and example 9 were placed in a stabilizing box with a temperature of 40.+ -. 2 ℃ and a relative humidity of 75% + -5% for 6 months, and were sampled at months 0, 1,3 and 6, respectively, and the contents of the three samples and the dissolution rate of 60min were measured, and the measurement results are shown in Table 11.
TABLE 11 accelerated stability test results
The accelerated stability test result shows that the sodium crocetin enteric preparation and the crocetin enteric preparation prepared by the invention have good stability under the accelerated condition, and the dissolution rate and the content are not obviously changed under the accelerated condition of six months through content measurement and detection of related substances and 60min dissolution rate, and all detection indexes meet the regulations.
3. Pharmacokinetic comparative experiments of enteric crocetin preparation
In order to observe the dissolution and absorption behaviors in different prescriptions, the drug substitution force experiment in the animal body is combined for verification in the prescription screening process.
3.1 Sample for sample
Example 2 sodium crocetin tablets prepared in prescription 1, sodium crocetin enteric-coated tablets prepared in examples 5, 7, 8 and 9, and sodium crocetin enteric-coated pellets prepared in example 10.
3.2, Laboratory animals
Common grade Beagle dogs, weight of 11.84-13.56kg, age of 16-24 months, male and female halves, and quality qualification number of experimental animals of No.110334210100040471 and No.370825220100019471. The temperature of the raising environment is 16-26 ℃ and the humidity is 40-70%. The day and night light and shade alternate time is 12h/12h. The animals in this batch were subjected to quarantine observation for at least 14 days in the specified feeding area of the center, observed 1 time a day, and the experiment was started after no obvious abnormality was found.
3.3 Grouping and dose design
The groups were divided into 6 groups of 4 to 6 male and female halves according to Table 12. The administration dosage is 100 mg/dog, and the administration mode is oral feeding.
TABLE 12 grouping dosing and dosage design
3.4 Animal administration and sample collection
The oral cavity and esophagus of the dogs are wetted with 15mL of purified water before each group of drugs is administered, then the drugs are placed at the tongue root of the dogs, and after the dogs swallow, the drugs are taken with 25mL of purified water. Blood samples were collected from the canine limb saphenous vein at corresponding time points before and after dosing in about 2mL of whole blood to the pre-labeled heparinized blood collection tube, and blood samples were collected once before and after dosing for each group of animals at 0.5h, 1h, 2h, 3h, 4h, 5h, 6h, 8h, 10h, 12h, 16h, 20 h.
3.5, Detection results
The calculation of each group of pharmacokinetic parameters was performed, and the specific parameters of the main pharmacokinetics of CRA in plasma after each administration group was shown in tables 13 and 14.
TABLE 13 results of drug-substitution parameter comparison study for example 5 sample group A animals
TABLE 14 results of drug parameter comparison study for example 2 prescription 1 sample group B animals
| Parameters (parameters) | AUC(0-t) | AUC(0-∞) | MRT(0-t) | MRT(0-∞) | t1/2 | Tmax | V | CL | Cmax |
| Unit (B) | μg/L*h | μg/L*h | h | h | h | h | L | L/h | μg/L |
| B1 | 5341.9 | 5343.4 | 11.655 | 11.659 | 1.727 | 4 | 18.652 | 7.486 | 598.6 |
| B2 | 3692.7 | 3692.7 | 4.123 | 4.123 | 0.955 | 4 | 14.924 | 10.832 | 936.2 |
| B3 | 2830.2 | 2832.7 | 6.834 | 6.838 | 1.618 | 3 | 7.668 | 5.854 | 766.7 |
| B4 | 1348.6 | 1348.6 | 5.458 | 5.458 | 0.912 | 2 | 9.844 | 4.479 | 498.8 |
| mean | 3303.4 | 3304.4 | 7.02 | 7.02 | 1.30 | 3.25 | 12.77 | 7.16 | 700.1 |
| SD | 1668.6 | 1668.9 | 3.28 | 3.29 | 0.43 | 0.96 | 4.96 | 2.74 | 192.4 |
As can be seen from the experimental results in tables 13 and 14, the enteric-coated tablets of example 5 of the same formulation and process had a bioavailability 3 times that of the uncoated plain tablets of example 2 formulation 1. Thus, to further verify the inventive, clinical value of the whole formulation of the present invention, beagle canine drug substitution comparison studies were performed according to the samples of examples 7, 8, 9, 10 and 5 in table 12, in conjunction with in vitro dissolution experiments.
Table 15. Summary of the results of the comparison study of the pharmacokinetic parameters of Beagle dogs for each example (n=6)
Based on the above results, the comparative examples of the conventional prescription CRAT tablets of example 2 show that the enteric-coated formulations of the present invention significantly improve the bioavailability of crocetin due to the lower AUC (0-t)、t1/2 and C max compared to the sodium crocetin enteric-coated formulations of examples 7-10. In addition, the sodium crocetin enteric coated tablet of example 5 without the pH regulator, the solubilizer and the lipid material and the sodium crocetin enteric coated tablet of example 7 without the lipid material can improve the bioavailability and half-life of the drug, which is consistent with the conclusion of example 1. The sodium crocetin enteric-coated tablet of example 8, the sodium crocetin enteric-coated tablet of example 9 and the sodium crocetin enteric-coated pellet of example 10, which contain the whole prescription, all significantly improve AUC (0-t) and t max. The experimental result shows that the bioavailability of the sodium crocetin enteric-coated preparation prepared by the invention is about 3-4 times of that of the prescription 1 of the example 2. Compared with gastric-soluble preparations, the enteric-coated preparation prepared by the invention can obviously accelerate absorption speed, effectively prolong the half life of the drug and improve the bioavailability in vivo under the same administration dosage; compared with the conventional enteric preparation, the invention can obviously improve the oral bioavailability of the insoluble crocetin preparation.
4. Pharmacodynamics comparison experiment of crocetin enteric preparation
4.1, Experimental sample
Group A, sodium crocetin bulk drug, provided by Xinglin traditional Chinese medicine science and technology (Guangzhou) Co., ltd;
Group B samples prepared in example 5;
Group C the whole enteric pellets prepared in example 10.
4.2, Laboratory animals and methods
SPF-grade male SD rats, weighing 170.6-258.1g, were subjected to coronary ligation except for 6 sham groups, and were routinely fed for 8 weeks after chronic heart failure (HFrEF) model operation in which the ejection fraction of the rats was reduced due to coronary ligation. At week 8, echocardiography was examined and animals with Ejection Fraction (EF) <50% were randomly divided into 4 groups, namely, a model control group, group A (sodium crocetin drug substance), group B (common sodium crocetin enteric-coated tablet), and group C (full Fang Xigong sodium crocetin enteric-coated pellet), each group of 6, each group being intragastric 1 time per day for 5 weeks.
4.3 Grouping and dose design
The total of 5 groups was divided according to Table 16, and 6 doses of the drug were shown in Table 16, each of which was administered by gavage at a dose of 10mL/kg body weight.
TABLE 16 grouping dosing and dosage design
| Group of | Test article | Dosage of | Administration mode | Number of animals |
| False operation group | Blank solvent | - | Oral administration | 6 Pieces of |
| Model control group | Blank solvent | - | Oral administration | 6 Pieces of |
| Group A | Sodium crocetin bulk drug | 11.3 | Oral administration | 6 Pieces of |
| Group B | Common sodium crocetin enteric-coated tablet | 11.3 | Oral administration | 6 Pieces of |
| Group C | Full Fang Xigong sodium florate enteric-coated pellet | 11.3 | Oral administration | 6 Pieces of |
4.4, Animal sample collection and detection
Detecting ejection fraction (Ejection Fraction, EF) in echocardiography at week 0 and week 6 of administration, collecting blood by sublingual vein after anaesthetizing HFrEF rat at week 5, standing at room temperature for about 1h, centrifuging at 4000rpm for 10min, separating serum, and detecting N-terminal brain natriuretic peptide precursor (NT-proBNP);
the heart of the animal was taken, pre-chilled saline was rinsed, filter paper was blotted dry, rapidly weighed, and heart index (CI) was calculated.
4.5, The detection results are as follows
NT-proBNP is a marker of myocardial damage, the content of NT-proBNP is proportional to the degree of myocardial damage, EF is called ejection fraction, the systolic and diastolic functions of the heart of a human body can be evaluated, and the effective ejection amount per minute can be evaluated, and the detection results of NT-proBNP, cardiac index and EF of each group of administration groups are shown in Table 17 and FIGS. 2 and 3.
TABLE 17 results of cardiac functional tests for each administration set
Note that vs sham group ##, p <0.01, vs model control group **, p <0.01;
As can be seen from Table 17, at week 5 of administration, the serum NT-proBNP content and cardiac index of the model control group were both increased (p < 0.01) and the serum NT-proBNP and cardiac index of group A, group B and group C were both decreased (p < 0.01) compared to the sham-operated group, indicating that crocetin could improve heart failure to some extent, protect myocardial cells, alleviate myocardial oxidative stress injury, and improve myocardial fibrosis degree. In addition, there was an increase in EF% after 5 weeks of administration (p < 0.01) in groups a, B and C compared to the model control group, indicating that crocetin improved cardiac ejection function in rats with heart failure, whereas the enteric pellets of group C had a significant effect of improving ejection function. From the results, the crocetin can be used for treating heart failure diseases, and the effect of the full Fang Xigong crocetin enteric-coated pellets of the group C containing the pH regulator, the solubilizer and the lipid material is obviously stronger than that of the common sodium crocetin enteric-coated tablets of the group B and the bulk drug of the group A, so that the full Fang Xigong crocetin enteric-coated pellets have the best treatment effect on HFrEF. The crocetin enteric preparation prepared by the invention can effectively prevent and treat cardiovascular and cerebrovascular diseases, and greatly expands the clinical value and the drug property of crocetin.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (3)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410811217.4A CN118845791B (en) | 2024-06-21 | 2024-06-21 | Crocetin preparation with high bioavailability and application thereof |
| PCT/CN2024/107289 WO2025260444A1 (en) | 2024-06-21 | 2024-07-24 | Crocetin preparation with high bioavailability, and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202410811217.4A CN118845791B (en) | 2024-06-21 | 2024-06-21 | Crocetin preparation with high bioavailability and application thereof |
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| Publication Number | Publication Date |
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| CN118845791B true CN118845791B (en) | 2025-06-27 |
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| CN113398077A (en) * | 2021-06-04 | 2021-09-17 | 浙江九如堂生物科技有限公司 | Composition for improving bioavailability of crocetin and oral preparation thereof |
| CN118924914A (en) * | 2024-01-18 | 2024-11-12 | 杏林中医药科技(广州)有限公司 | A sodium crocetin nanocomplex preparation with high bioavailability, preparation method, medicine and application |
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| CN113398077A (en) * | 2021-06-04 | 2021-09-17 | 浙江九如堂生物科技有限公司 | Composition for improving bioavailability of crocetin and oral preparation thereof |
| CN118924914A (en) * | 2024-01-18 | 2024-11-12 | 杏林中医药科技(广州)有限公司 | A sodium crocetin nanocomplex preparation with high bioavailability, preparation method, medicine and application |
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