CN110882398A - Oral curcumin-nano diamond compound and preparation method thereof - Google Patents
Oral curcumin-nano diamond compound and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of pharmaceutical preparations, and discloses an oral curcumin-nanodiamond compound and a preparation method thereof. According to the invention, firstly, the nano-diamond is subjected to carboxylation and acylchlorination treatment in sequence, then the acylchlorinated nano-diamond is reacted with polyethylene glycol vitamin E succinate to obtain polyethylene glycol vitamin E succinate covalently modified nano-diamond, and then curcumin is entrapped to obtain the curcumin nano-diamond compound. The covalent modification of polyethylene glycol vitamin E succinate effectively solves the problems of dispersibility and in-vivo and in-vitro stability of the nanodiamond serving as a drug delivery carrier, obviously improves the solubility, gastrointestinal retention and permeability of the drug after the curcumin is entrapped, and obviously improves the oral bioavailability of the drug.
Description
The technical field is as follows:
the invention belongs to the field of pharmaceutical preparations, and relates to an oral curcumin-nanodiamond compound and a preparation method thereof.
Background art:
nanodiamonds (NDs) are carbon materials with a truncated octahedral structure, and the particle size of a single particle is 2 to 8 nm. The NDs has the advantages of small size, large specific surface area, stable chemical property, good biocompatibility, easy surface modification and the like, so that the NDs becomes a novel drug delivery carrier with great application prospect. NDs spontaneously form nano-sized clusters in aqueous solution, and the surface of the NDs is rich in a large number of oxygen-containing groups, such as hydroxyl, carboxyl, lactone and the like, so that the drugs can be bound to the surface of the NDs or embedded in the clusters by physical adsorption or covalent connection. Many evidences show that NDs serving as a drug carrier can enhance the cellular uptake of the drug, prolong the retention time in vivo and improve the bioavailability and the medication safety of the drug.
While clustering of NDs is beneficial for drug loading, excessive clustering can reduce the colloidal stability of the formulation, hinder gastrointestinal transport of the carrier, and trigger some cytotoxicity. Although redispersion of NDs can be achieved by sonication, mechanical milling, etc. after addition of a certain amount of surfactant to the solvent, there is still a risk of the NDs re-aggregating during the placement of the formulation, and in the complex physiological environment in the body. In contrast, surface chemical modification is more effective in improving dispersion stability of NDs.
Polyethylene glycol vitamin E succinate (TPGS) is a water-soluble derivative of vitamin E, the amphipathy and the larger molecular surface area of the TPGS enable the TPGS to be an excellent nonionic surfactant, the critical micelle concentration is only 0.02%, the polar PEG head and the nonpolar TOS tail of the TPGS both have huge surface areas, the acting force of the TPGS on an oil-water two-phase is stronger, and the TPGS can be soluble in water and most organic solvents.
Curcumin (curmin, CUR) is a polyphenol substance extracted from rhizomes of plants in the genus of zingiberaceae, has wide source and low toxicity, and has various pharmacological activities such as anti-inflammatory, antioxidant and antitumor. However, CUR is hardly soluble in water (solubility is only 11ng/mL in ph5.0 buffer), degrades rapidly under neutral, alkaline and illumination conditions, is mostly excreted out of the body through feces after oral administration, is rapidly metabolized in the liver and blood after being absorbed by the gastrointestinal tract for a very small part, has obvious first pass effect, has only 1% of oral absolute bioavailability, and belongs to class iv drugs in the Biopharmaceutical Classification System (BCS). How to effectively improve the oral bioavailability of the CUR is a key scientific problem to be urgently solved in the pharmaceutical science community.
Therefore, TPGS is grafted on the surface of NDs in a covalent connection mode to solve the problems of colloidal dispersibility and dispersion stability of NDs, and the CUR is coated in NDs-TPGS nanoclusters in an amorphous state to obtain a CUR @ NDs-TPGS nano compound, so that the solubility, gastrointestinal tract adhesion, permeability and oral bioavailability of the CUR are greatly improved, and the problem of drug forming property of the CUR is solved.
The invention content is as follows:
the first purpose of the invention is to provide a method for synthesizing NDs-TPGS, which enables a product to have higher grafting ratio of TPGS, smaller particle size and good dispersibility by optimizing synthesis conditions, thereby obtaining a drug delivery carrier with wide application prospect.
The invention aims to provide a prescription of a CUR @ NDs-TPGS nano compound and a preparation method thereof, wherein the medicine carrying efficiency, the particle size distribution and the like of the preparation are controlled within a reasonable range by optimizing the prescription process, so that the solubility and the oral bioavailability of the CUR are improved, and the problem of the pharmacy of the CUR is solved.
The invention is realized by the following technical scheme:
carrying out carboxylation and acyl chlorination treatment on NDs in sequence, and further reacting the acyl chlorinated NDs with TPGS to synthesize an NDs-TPGS nano-drug carrier; and (3) encapsulating the CUR in a nanocage formed by NDs-TPGS to obtain the CUR @ NDs-TPGS nano compound.
The NDs-TPGS synthesis method comprises the following steps:
(1) NDs is added into a certain volume of mixed acid at a certain temperature, stirred for a certain time, cooled to room temperature, washed with water and centrifuged; adjusting the pH value to 5-6 by NaOH and HCl; and (4) washing with water, and drying in vacuum to obtain NDs-COOH.
(2) NDs-COOH is ultrasonically dispersed in oxalyl chloride, then dimethylformamide is added, stirring and reacting are carried out for a certain time at a certain temperature, NDs-COCl is washed by anhydrous tetrahydrofuran after centrifugation, and vacuum drying is carried out, so as to obtain the NDs-COCl.
(3) And ultrasonically dispersing NDs-COCl in dimethylformamide, adding TPGS to react for a certain time at a certain temperature, washing the cooled reaction solution with water, and freeze-drying to obtain the NDs-TPGS.
The mixed acid in the step (1) is H2SO4-HNO3、H2SO4-HClO4And H2SO4-one of HCl in a volume ratio of 5:1 to 1:2, preferably in a range of 5:1 to 2: 1.
The reaction temperature of the NDs in the step (1) in the mixed acid is 25-100 ℃.
The concentration of the NDs in the mixed acid in the step (1) is 5-55 mg/mL, and the preferable concentration range is 5-40 mg/mL.
The reaction time of the NDs in the step (1) in the mixed acid is 12-48 h.
The concentration of the NDs-COOH in the oxalyl chloride in the step (2) is 2-16 mg/mL, and the preferable concentration range is 2-8 mg/mL.
The concentration of the dimethylformamide in the step (2) is 0.2-1 mg/mL.
The reaction time in the step (2) is 6-48 h.
The reaction temperature in the step (2) is 25-75 ℃, and the preferable temperature is 40-75 ℃.
The concentration of the NDs-COCl in the dimethylformamide in the step (3) is 0.1-10 mg/mL, and the preferable range is 0.1-5 mg/mL.
The concentration of the TPGS in the dimethylformamide in the step (3) is 1-20 mg/mL, and the preferable range is 3-20 mg/mL.
The TPGS in the step (3) is of the type of TPGS400、TPGS1000、TPGS2000、TPGS4000And TPGS6000One kind of (1).
The reaction time in the step (3) is 12-72 h.
The reaction temperature in the step (3) is 40-120 ℃.
Carrying out comprehensive optimization screening by taking the grafting rate, the particle size and the particle size distribution as evaluation indexes, wherein when the concentration of the NDs in the step (1) is 5-30 mg/mL; and the concentration of NDs-COOH in the step (2) is 2-8 mg/mL; and (3) when the concentration of TPGS in the step (3) is 2-10 mg/mL, the excellent nano-carrier with the grafting rate of more than 45%, the particle size of less than 250nm and the particle size distribution of less than 0.25 can be obtained.
The preparation method of the CUR @ NDs-TPGS nano compound comprises the following steps:
(1) adding a certain amount of NDs-TPGS into a proper solvent, and ultrasonically dispersing for a certain time at a certain temperature by using a probe with certain power.
(2) The formula amount of CUR was dissolved in a suitable solvent.
(3) And (3) dripping the CUR solution into the NDs-TPGS suspension, continuing probe ultrasonic treatment (the power is consistent with that in the step (1)), and drying under reduced pressure to remove the organic solvent to obtain the CUR @ NDs-TPGS.
(4) And adding distilled water into the CUR @ NDs-TPGS, and carrying out redispersion by using probe ultrasound to obtain the product.
The concentration of the NDs-TPGS in the step (1) is 1-30 mg/mL, and the preferable concentration range is 5-30.
The NDs-TPGS in the step (1) is NDs-TPGS400、NDs-TPGS1000、NDs-TPGS2000、NDs-TPGS4000One kind of (1).
The dispersion solvent in the step (1) is one of ethanol, 50% ethanol, 75% ethanol, dimethyl sulfoxide, dimethylformamide, acetone and dichloromethane.
The ultrasonic power in the step (1) is 120-600W, and preferably 240-600W.
The ultrasonic time in the step (1) is 5-60 min, and the preferable ultrasonic time is 15-60 min.
The temperature in the step (1) is 0-50 ℃.
The concentration of the CUR in the step (2) is 0.5-3 mg/mL, and the preferable concentration range is 0.5-2 mg/mL.
And (3) the solvent in the step (2) is one of ethanol, dimethyl sulfoxide, dimethylformamide, acetone and dichloromethane.
Comprehensively optimizing a preparation prescription by taking the drug loading efficiency, the particle size and the particle size distribution as evaluation indexes, wherein when the concentration of the NDs-TPGS in the step (1) is 5-20 mg/mL, the NDs-TPGS is NDs-TPGS400、NDs-TPGS1000Or NDs-TPGS2000One of (1); and when the CUR concentration in the step (2) is 0.5-2 mg/mL, the drug loading efficiency of the obtained preparation is more than 50%, the particle size is less than 350nm, and the particle size distribution is less than 0.3.
The invention has the following beneficial effects:
1. the NDs-TPGS synthesized by the method can obviously improve the dispersibility and dispersion stability of the NDs; compared with the conventional physical coating method, the NDs modified by TPGS through a covalent linkage method can further improve the in vivo and in vitro stability and the drug delivery efficiency of the carrier.
2. Compared with a naked NDs-COOH (CUR @ NDs-COOH) and TPGS (CUR @ NDs-COOH/TPGS) physical coating NDs system, after the NDs-TPGS coats the CUR, the particle size of a drug delivery system is reduced, the particle size distribution is more uniform, the drug loading efficiency is improved, the intestinal cell uptake is obviously increased, the distribution, the retention and the permeability of the preparation in the gastrointestinal tract of a rat are further improved, the oral bioavailability is obviously improved, and the sustained release effect is good.
Description of the drawings:
FIG. 1NDs-TPGS Synthesis roadmap
FIG. 2CUR @ NDs-TPGS Transmission Electron microscope picture and structure schematic diagram
FIG. 3 is an X-ray diffraction pattern of CUR @ NDs-TPGS prepared according to example 1
FIG. 4 intestinal Retention study of NDs-TPGS (small animal in vivo imaging plot) (a) coumarin 6(Cou-6) solution, (b) Cou-6@ NDs-COOH/TPGS, (c) Cou-6@ NDs-TPGS
FIG. 5 is a graph showing the time course of administration of the CUR suspension, CUR @ NDs-COOH/TPGS and CUR @ NDs-TPGS after gastric gavage in rats at a dose of 75 mg/kg.
Detailed Description
The present invention will be described in further detail with reference to examples, but the scope of the present invention is not limited by these examples.
EXAMPLE 1 screening of NDs-TPGS Synthesis conditions
(1) NDs is added into mixed acid with a certain volume at a certain temperature, stirred for a certain time, cooled to room temperature, diluted by distilled water and centrifuged; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
(2) 100mg of NDs-COOH was ultrasonically dispersed in 25mL of oxalyl chloride, then 1mL of dimethylformamide was added, the mixture was stirred at a certain temperature and reacted for 24 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried in vacuum for 24 hours for later use.
(3) 10mg of NDs-COCl was ultrasonically dispersed in 10mL of dimethylformamide, followed by the addition of 50mg of TPGS1000Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
After the synthesis is finished, respectively detecting the weight loss conditions of NDs-COOH, TPGS and NDs-TPGS in the temperature range of 25-550 ℃ by using an Shimadzu TGA-50 thermogravimetric analyzer, and calculating the grafting ratio; 0.05g of NDs-TPGS was weighed, subjected to ice-bath ultrasonic treatment for 30min (ultrasonic power 300w), dispersed in 10mL of distilled water, and the particle size and particle size distribution were measured using a Malvern Zetasizer particle sizer. The best synthesis conditions are that the grafting rate is more than 50 percent, the grain diameter is less than 250nm, and the grain size distribution is less than 0.25; using 30-50% of grafting rate, 250-300 nm of particle size and 0.25-0.3 of particle size distribution as suboptimal synthesis conditions; the screening synthesis process is carried out under the conditions of graft rate less than 30%, particle size greater than 300nm and particle size distribution greater than 0.3. The result shows that the NDs has the best synthetic effect when the concentration range of the NDs in the mixed acid is 5-25 mg/mL, the best NDs are obtained when the concentration range of the NDs is 25-40 mg/mL, the average NDs effect is 40-55 mg/mL, and the poor NDs effect is 55-70 mg/mL; when the mixed acid is H2SO4-HNO3And H2SO4Best synthesis when HCl is used, H2SO4-HClO4The effect is the next time; when the mixed acid ratio is 5: 1-3: 1, the synthesis effect is optimal, the ratio is 3: 1-2: 1, the effect is inferior, and the effect is common when the ratio is 2: 1-1: 2; the reaction time in the mixed acid is 12-48 hThe effect is optimal; the effect is best when the reaction temperature is 40-100 ℃, and the effect is sub-best when the reaction temperature is 25-40 ℃.
TABLE 1 Effect of carboxylation reaction conditions on NDs-TPGS grafting Rate, particle size and particle size distribution
EXAMPLE 2 screening of NDs-TPGS Synthesis conditions
(1) 0.5g of NDs was added to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
(2) Ultrasonically dispersing a certain amount of NDs-COOH in 25mL of oxalyl chloride, then adding a certain amount of dimethylformamide, stirring and reacting for a certain time at a certain temperature, centrifuging, washing NDs-COCl with anhydrous tetrahydrofuran for three times, and drying in vacuum for 24 hours for later use.
(3) 10mg of NDs-COCl was ultrasonically dispersed in 10mL of dimethylformamide, followed by the addition of 50mg of TPGS1000Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
After the synthesis is finished, respectively detecting the weight loss conditions of NDs-COOH, TPGS and NDs-TPGS in the temperature range of 25-550 ℃ by using an Shimadzu TGA-50 thermogravimetric analyzer, and calculating the grafting ratio; 0.05g of NDs-TPGS was weighed, subjected to ice-bath ultrasonic treatment for 30min (ultrasonic power 300w), dispersed in 10mL of distilled water, and the particle size and particle size distribution were measured using a Malvern Zetasizer particle sizer. The best synthesis conditions are that the grafting rate is more than 50 percent, the grain diameter is less than 250nm, and the grain size distribution is less than 0.25; using 30-50% of grafting rate, 250-300 nm of particle size and 0.25-0.3 of particle size distribution as suboptimal synthesis conditions; the screening synthesis process is carried out under the conditions of graft rate less than 30%, particle size greater than 300nm and particle size distribution greater than 0.3. The result shows that the NDs-COOH has the best synthesis effect when the concentration of the NDs-COOH in oxalyl chloride is 2-8 mg/mL, and the best synthesis effect is not obtained when the concentration of the NDs-COOH in oxalyl chloride is 8-16 mg/mL; the best synthetic effect is achieved when the concentration of the dimethylformamide is 0.2-1 mg/mL; the reaction time is 12 to 48 hours, the effect is the best, and the effect is the second time after 6 to 12 hours; the reaction temperature is 40-75 ℃ with the best effect, and the reaction temperature is 25-40 ℃ with the ordinary effect.
TABLE 2 influence of NDs-COCl Synthesis conditions on the grafting ratio, particle size and particle size distribution of NDs-TPGS
EXAMPLE 3 screening of NDs-TPGS Synthesis conditions
(1) 0.5g of NDs was added to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
(2) 100mg of NDs-COOH was ultrasonically dispersed in 25mL of oxalyl chloride, then 1mL of dimethylformamide was added, the mixture was stirred at a certain temperature and reacted for 24 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried in vacuum for 24 hours for later use.
(3) Ultrasonically dispersing a certain amount of NDs-COCl into 10mL of dimethylformamide, then adding a certain amount of TPGS to react for a certain time at a certain temperature, filtering the cooled reaction solution by using a 0.22 mu m microporous filter membrane, washing with water, and freeze-drying to obtain the NDs-TPGS.
After the synthesis is finished, respectively detecting the weight loss conditions of NDs-COOH, TPGS and NDs-TPGS in the temperature range of 25-550 ℃ by using an Shimadzu TGA-50 thermogravimetric analyzer, and calculating the grafting ratio; 0.05g of NDs-TPGS was weighed, subjected to ice-bath ultrasonic treatment for 30min (ultrasonic power 300w), dispersed in 10mL of distilled water, and the particle size and particle size distribution were measured using a Malvern Zetasizer particle sizer. The best synthesis conditions are that the grafting rate is more than 50 percent, the grain diameter is less than 250nm, and the grain size distribution is less than 0.25; the grafting rate is 30-50%, the particle size is 250-300 nm, and the particle size isCloth 0.25-0.3 is a sub-optimal synthesis condition; the screening synthesis process is carried out under the conditions of graft rate less than 30%, particle size greater than 300nm and particle size distribution greater than 0.3. The result shows that the NDs-COCl has the best synthetic effect when the concentration of the NDs-COCl in the dimethylformamide is 0.1-5 mg/mL, and the effect is general when the concentration of the NDs-COCl in the dimethylformamide is 5-10 mg/mL; the effect of TPGS in dimethylformamide is the best when the concentration of TPGS is 3-20 mg/mL, and the effect is sub-optimal when the concentration of TPGS in dimethylformamide is 1-3 mg/mL; the TPGS is TPGS400、TPGS1000And TPGS2000The best effect is TPGS4000The second best effect is TPGS6000The effect is general; the reaction time is 12-72 h, and the result is good; the effect is best when the reaction temperature is 60-120 ℃, and the effect is sub-best when the reaction temperature is 40-60 ℃.
TABLE 3 influence of NDs-TPGS Synthesis conditions on grafting Rate, particle size and particle size distribution
Example 4 optimization of the Synthesis conditions of NDs-TPGS
Determining three key factors influencing the NDs-TPGS synthesis as the NDs concentration in the step (1), the NDs-COOH concentration in the step (2) and the TPGS concentration in the step (3) by single factor investigation, and further optimizing the synthesis conditions:
(1) a defined amount of NDs was added to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
(2) Ultrasonically dispersing a certain amount of NDs-COOH in 25mL of oxalyl chloride, then adding 0.5mL of dimethylformamide, stirring and reacting at 60 ℃ for 24h, centrifuging, washing NDs-COCl with anhydrous tetrahydrofuran three times, and drying in vacuum for 24h for later use.
(3) Ultrasonically dispersing 10mg of NDs-COCl in 10mL of dimethylformamide, and then adding a certain amount of TPGS2000Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
After the synthesis is finished, respectively detecting the weight loss conditions of NDs-COOH, TPGS and NDs-TPGS in the temperature range of 25-550 ℃ by using an Shimadzu TGA-50 thermogravimetric analyzer, and calculating the grafting ratio; 0.05g of NDs-TPGS was weighed, subjected to ice-bath ultrasonic treatment for 30min (ultrasonic power 300w), dispersed in 10mL of distilled water, and the particle size and particle size distribution were measured using a Malvern Zetasizer particle sizer. Calculating the average value of different level indexes of different factors, calculating the difference (range) between the lowest value and the highest value of the average value, and evaluating the significance degree of the factors (see table 5).
According to the investigation result, the obvious influence degrees of all factors on the grafting rate, the particle size and the particle size distribution of the synthesized product NDs-TPGS are as follows: step (3) TPGS concentration > step (2) NDs-COOH concentration > step (1) NDs concentration. When the concentration of the NDs in the step (1) is 5-30 mg/mL; and the concentration of NDs-COOH in the step (2) is 2-8 mg/mL; and (3) when the concentration of TPGS in the step (3) is 2-10 mg/mL, the excellent nano-carrier with the grafting rate of more than 45%, the particle size of less than 250nm and the particle size distribution of less than 0.25 can be obtained.
TABLE 4 optimization of NDs-TPGS Synthesis conditions
TABLE 5 analysis of optimization results
Example 5 prescription screening of CUR @ NDs-TPGS nanocomposites
Synthesis method of NDs-TPGS
1.1 Add 0.5g of NDs to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
1.2 ultrasonic dispersion of 100mg NDs-COOH in 25mL oxalyl chloride, then adding 1mL dimethylformamide, stirring at a certain temperature for reaction for 24 hours, centrifuging, washing NDs-COCl with anhydrous tetrahydrofuran three times, and vacuum drying for 24 hours for later use.
1.3 ultrasonic Dispersion of 10mg of NDs-COCl in 10mL of dimethylformamide followed by addition of 50mg of TPGS1000Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
Preparation method of CUR @ NDs-TPGS nano compound
2.1A quantity of NDs-TPGS1000Adding into 10mL of proper solvent, and dispersing for 30min by probe ultrasonic (360w, 0 ℃).
2.2 dissolve the prescription amount of CUR in 2mL of the appropriate solvent.
2.3 dropping the CUR solution into NDs-TPGS1000And (4) continuing probe ultrasonic treatment for 30min (the power is consistent with the step 2.1), and drying under reduced pressure to remove the organic solvent.
2.4 Direction CUR @ NDs-TPGS1000Adding 10mL of distilled water, and carrying out redispersion by probe ultrasonic treatment (360w, 0 ℃) for 10min to obtain the product.
The drug loading efficiency of the preparation is determined by ultraviolet-visible spectrophotometry, and the particle size distribution are determined by a Malvern Zetasizer. The optimal conditions are that the drug loading efficiency is more than 70%, the particle size is less than 300nm, and the particle size distribution is less than 0.3; the drug loading efficiency is 50-70%, the particle size is 300-400 nm, and the particle size distribution is 0.3-0.35, which is a sub-optimal condition; the drug loading efficiency is less than 50%, the particle size is more than 400nm, and the particle size distribution is more than 0.35, which is the screening prescription under the common conditions. The result shows that the NDs-TPGS has the best effect when the concentration is 5-20 mg/mL, the effect is not good when the concentration is 3-5 mg/mL and 20-30 mg/mL, and the effect is general when the concentration is 1-3 mg/mL; the effect is best when the concentration of the CUR is 0.5-1 mg/mL, the effect is sub-optimal when the concentration of the CUR is 1-2 mg/mL, and the effect is common when the concentration of the CUR is 2-3 mg/mL; in the step 2.1, the NDs-TPGS dispersion solvent has the best effect when being one of acetone, dimethylformamide, ethanol and 75% ethanol, has the second best effect when being dimethyl sulfoxide or 50% ethanol, and has the common effect when being dichloromethane; the solvent of step 2.2 works best with acetone or ethanol, less well with dimethyl sulfoxide or dimethylformamide, and generally with dichloromethane.
TABLE 6 prescription screening of CUR @ NDs-TPGS nanocomposites
Example 6 Process screening of CUR @ NDs-TPGS nanocomposites
Synthesis method of NDs-TPGS
1.1 Add 0.5g of NDs to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
1.2 ultrasonic dispersion of 100mg NDs-COOH in 25mL oxalyl chloride, then adding 1mL dimethylformamide, stirring at a certain temperature for reaction for 24 hours, centrifuging, washing NDs-COCl with anhydrous tetrahydrofuran three times, and vacuum drying for 24 hours for later use.
1.3 ultrasonic Dispersion of 10mg of NDs-COCl in 10mL of dimethylformamide followed by addition of 50mg of TPGS1000Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
Preparation method of CUR @ NDs-TPGS nano compound
2.1 Add 50mg of NDs-TPGS1000Adding into 10mL ethanol, and ultrasonically dispersing for a certain time at a certain temperature by a probe with certain power.
2.2 dissolve 3mg of CUR in 2mL of acetone.
2.3 dropping the CUR solution into NDs-TPGS1000And (4) continuing to perform ultrasonic treatment for 30min by a probe with certain power (the power is consistent with that in the step 2.1), and decompressing and drying to remove the organic solvent.
2.4 Direction CUR @ NDs-TPGS1000Adding 10mL of distilled water, and carrying out redispersion by probe ultrasonic treatment (360w, 0 ℃) for 10min to obtain the product.
The drug loading efficiency of the preparation is determined by ultraviolet-visible spectrophotometry, and the particle size distribution are determined by a Malvern Zetasizer. The optimal conditions are that the drug loading efficiency is more than 70%, the particle size is less than 300nm, and the particle size distribution is less than 0.3; the drug loading efficiency is 50-70%, the particle size is 300-400 nm, and the particle size distribution is 0.3-0.35, which is a sub-optimal condition; the screening process conditions under the common conditions are that the drug loading efficiency is less than 50 percent, the particle size is more than 400nm, and the particle size distribution is more than 0.35. The result shows that the ultrasonic power is 360-600W, the effect is the best, 240-360W is the next best, and 120-240W is the general effect; the ultrasonic temperature is 0-25 ℃ for the best effect, and the effect is inferior at 25-50 ℃; the best effect is obtained when the ultrasonic time is 15-60 min, and the next effect is obtained when the ultrasonic time is 5-15 min.
TABLE 7 Process screening of CUR @ NDs-TPGS nanocomposites
Example 7 Effect of model TPGS on nanocomposite formulation Properties
Synthesis method of NDs-TPGS
1.1 Add 0.5g of NDs to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
1.2 ultrasonic dispersion of 100mg NDs-COOH in 25mL oxalyl chloride, then adding 1mL dimethylformamide, stirring at a certain temperature for reaction for 24 hours, centrifuging, washing NDs-COCl with anhydrous tetrahydrofuran three times, and vacuum drying for 24 hours for later use.
1.3 sonication of 10mg of NDs-COCl in 10mL of dimethylformamide followed by the addition of 50mg of TPGS (TPGS)400Or TPGS1000Or TPGS2000Or TPGS2000) Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
Preparation method of CUR @ NDs-TPGS nano compound
2.1 Add 50mg of NDs-TPGS to 10mL of ethanol and disperse by probe sonication (360w, 0 ℃) for 30 min.
2.2 dissolve 3mg of CUR in 2mL of acetone.
2.3 dripping the CUR solution into the NDs-TPGS suspension, continuing to perform ultrasonic treatment for 30min by a probe with certain power (the power is consistent with the power in the step 2.1), and drying under reduced pressure to remove the organic solvent.
2.4 adding 10mL of distilled water into the CUR @ NDs-TPGS, and carrying out redispersion for 10min by probe ultrasound (360w, 0 ℃ C.).
The drug loading efficiency of the preparation is determined by ultraviolet-visible spectrophotometry, and the particle size distribution are determined by a Malvern Zetasizer. The optimal conditions are that the drug loading efficiency is more than 70%, the particle size is less than 300nm, and the particle size distribution is less than 0.3; the drug loading efficiency is 50-70%, the particle size is 300-400 nm, and the particle size distribution is 0.3-0.35, which is a sub-optimal condition; the TPGS model is screened under the general conditions that the drug loading efficiency is less than 50 percent, the particle size is more than 400nm and the particle size distribution is more than 0.35. As a result, TPGS was obtained under the same process and formulation conditions1000And TPGS2000Is best in effect, TPGS400The second effect of (2), TPGS4000The effect of (2) is general.
TABLE 8 Effect of TPGS type on nanocomposite formulation Properties
Example 8CUR @ NDs-TPGS nanocomposite prescription optimization
Synthesis method of NDs-TPGS
1.1 Add a quantity of NDs to 20mL of mixed acid (H) at 60 deg.C2SO4-HCl in a volume ratio of 3:1), stirring for 24 hours, cooling to room temperature, diluting with distilled water, and centrifuging; adjusting the pH value to 5-6 by using 0.1M NaOH and 0.1M HCl; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
1.2 ultrasonic dispersion of 100mg NDs-COOH in 25mL oxalyl chloride, then adding 1mL dimethylformamide, stirring at a certain temperature for reaction for 24 hours, centrifuging, washing NDs-COCl with anhydrous tetrahydrofuran three times, and vacuum drying for 24 hours for later use.
1.3 sonication of 10mg of NDs-COCl in 10mL of dimethylformamide followed by the addition of 50mg of TPGS (TPGS)400Or TPGS1000Or TPGS2000Or TPGS2000) Reacting at 100 deg.C for 48 hr, filtering the cooled reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS.
Preparation method of CUR @ NDs-TPGS nano compound
2.1A certain amount of NDs-TPGS was added to 10mL of ethanol and dispersed by ultrasonic probe (360w, 0 ℃) for 30 min.
2.2 dissolve an amount of CUR in 2mL of acetone.
2.3 dripping the CUR solution into the NDs-TPGS suspension, continuing to perform ultrasonic treatment for 30min by using a probe, and drying under reduced pressure to remove the organic solvent.
2.4 adding 10mL of distilled water into the CUR @ NDs-TPGS, and carrying out redispersion for 10min by probe ultrasound (360w, 0 ℃ C.).
The drug loading efficiency of the preparation is determined by ultraviolet-visible spectrophotometry, and the particle size distribution are determined by a Malvern Zetasizer. As can be seen from the optimization results, the significant influence of all factors on the drug loading efficiency, the particle size and the particle size distribution is sequentially the type of NDs-TPGS in the step 2.1>Step 2.2CUR concentration>Step 2.1NDs-TPGS concentration. When the concentration of the NDs-TPGS in the step (1) is 5-20 mg/mL, the NDs-TPGS is NDs-TPGS400、NDs-TPGS1000Or NDs-TPGS2000One of (1); and when the CUR concentration in the step (2) is 0.5-2 mg/mL, the drug loading efficiency of the obtained preparation is more than 50%, the particle size is less than 350nm, and the particle size distribution is less than 0.3.
TABLE 9CUR @ NDs-TPGS nanocomposite recipe optimization
TABLE 10 analysis of optimization results
Example 9 preparation of CUR @ NDs-TPGS oral Nanocomposite
(1) NDs-TPGS synthesis method
1) 0.5g of NDs was added to 20mL of mixed acid (H) at 60 deg.C2SO4-HNO3Stirring for 24 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging at 10,000rpm for 15 min; heating NDs particles in 0.1M NaOH at 90 ℃ for 2 hours, and heating in 0.1M HCl at 90 ℃ for 2 hours to reduce the pH value to 5-6; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
2) 0.1g of NDs-COOH was dispersed in 20mL of oxalyl chloride by sonication, 0.5mL of dimethylformamide was added, the reaction was stirred at 60 ℃ for 24 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried in vacuum for 24 hours.
3) 10mg of NDs-COCl was ultrasonically dispersed in 10mL of dimethylformamide, followed by the addition of 50mg of TPGS1000Reacting at 100 deg.C for 48 hr, filtering the reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS1000。
(2) Preparation method of CUR @ NDs-TPGS
1) 50mg of NDs-TPGS1000Dispersed in 10mL ethanol by probe ultrasound (360w, 0 ℃) for 30 min.
2) 3mg of CUR was dissolved in 2mL of acetone.
3) Dropwise adding the CUR solution to NDs-TPGS under ultrasound1000Ultrasonic treating in ethanol dispersion for 30min, and drying under reduced pressure.
4) 50mg of CUR @ NDs-TPGS1000Redispersed in 10mL of distilled water under ultrasound for 10 min.
The resulting CUR @ NDs-TPGS1000The average particle size is 196.33nm, the particle size distribution is 0.19, the potential is-24.52 mV, and the drug loading efficiency is 87.19%.
Example 10 preparation of CUR @ NDs-TPGS oral Nanocomposite
(1) NDs-TPGS synthesis method
1) 0.1g of NDs was added to 20mL of mixed acid (H) at 25 deg.C2SO4-HCl in a volume ratio of 2:1), stirring for 48 hours, cooling to room temperature, diluting with distilled water, and centrifuging at 10,000rpm for 15 min; NDs particles were at 0.Heating the mixture in 1M NaOH at 90 ℃ for 2 hours, and heating the mixture in 0.1M HCl at 90 ℃ for 2 hours to reduce the pH value to 5-6; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
2) 0.32g of NDs-COOH was dispersed in 20mL of oxalyl chloride by sonication, 1mL of dimethylformamide was added, the reaction was stirred at 75 ℃ for 48 hours, and NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation and dried under vacuum for 24 hours.
3) 100mg of NDs-COCl was dispersed in 10mL of dimethylformamide by sonication, followed by the addition of 30mg of TPGS2000Reacting at 120 deg.C for 72 hr, filtering the reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS2000。
(2) Preparation method of CUR @ NDs-TPGS
1) 300mg of NDs-TPGS2000Dispersed in 10mL 75% ethanol by probe ultrasound (600w, 25 ℃) for 60 min.
2) 4mg of CUR was dissolved in 2mL of dimethyl sulfoxide.
3) Dropwise adding the CUR solution to NDs-TPGS under ultrasound2000Ultrasonic treating in the dispersion for 30min, and drying under reduced pressure.
4) 50mg of CUR @ NDs-TPGS400Redispersed in 10mL of distilled water under ultrasound for 10 min.
The resulting CUR @ NDs-TPGS2000The average particle size is 368.07nm, the particle size distribution is 0.37, and the drug loading efficiency is 38.25%.
EXAMPLE 11 preparation of CUR @ NDs-TPGS oral Nanocomposite
(1) NDs-TPGS synthesis method
1) 1.1g of NDs was added to 20mL of mixed acid (H) at 60 deg.C2SO4-HCl in a volume ratio of 5:1), stirring for 12 hours, cooling to room temperature, diluting with distilled water, and centrifuging at 10,000rpm for 15 min; heating NDs particles in 0.1M NaOH at 90 ℃ for 2 hours, and heating in 0.1M HCl at 90 ℃ for 2 hours to reduce the pH value to 5-6; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
2) 0.04g of NDs-COOH was ultrasonically dispersed in 20mL of oxalyl chloride, 0.2mL of dimethylformamide was added, the reaction was stirred at 40 ℃ for 6 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried under vacuum for 24 hours.
3) 1mg of NDs-COCl was dispersed in 10mL of dimethylformamide by sonication, followed by the addition of 200mg of TPGS400Reacting at 100 deg.C for 12 hr, filtering the reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS400。
(2) Preparation method of CUR @ NDs-TPGS
1) 10mg of NDs-TPGS400Dispersed in 10mL 75% ethanol by probe ultrasound (120w, 50 ℃) for 10 min.
2) 1mg of CUR was dissolved in 2mL of ethanol.
3) Dropwise adding the CUR solution to NDs-TPGS under ultrasound400Ultrasonic treating in the dispersion for 30min, and drying under reduced pressure.
4) 50mg of CUR @ NDs-TPGS400Redispersed in 10mL of distilled water under ultrasound for 10 min.
The resulting CUR @ NDs-TPGS400The average particle size is 287.35nm, the particle size distribution is 0.28, and the drug loading efficiency is 60.27%.
Example 12 preparation of CUR @ NDs-TPGS oral Nanocomposite
(1) NDs-TPGS synthesis method
1) 0.3g of NDs was added to 20mL of mixed acid (H) at 100 deg.C2SO4-HClO4Stirring for 48 hours in a volume ratio of 3:1), cooling to room temperature, diluting with distilled water, and centrifuging at 10,000rpm for 15 min; heating NDs particles in 0.1M NaOH at 90 ℃ for 2 hours, and heating in 0.1M HCl at 90 ℃ for 2 hours to reduce the pH value to 5-6; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
2) 0.16g of NDs-COOH was dispersed in 20mL of oxalyl chloride by sonication, 0.5mL of dimethylformamide was added, the reaction was stirred at 90 ℃ for 36 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried in vacuum for 24 hours.
3) 50mg of NDs-COCl was dispersed in 10mL of dimethylformamide by sonication, followed by the addition of 100mg of TPGS4000Reacting at 120 deg.C for 72 hr, and collecting the reaction solutionFiltering with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS4000。
(2) Preparation method of CUR @ NDs-TPGS
1) 100mg of NDs-TPGS4000Dispersed in 10mL of dimethylformamide with probe ultrasound (240w, 36 ℃) for 20 min.
2) 5mg of CUR was dissolved in 2mL of dimethylformamide.
3) Dropwise adding the CUR solution to NDs-TPGS under ultrasound4000Ultrasonic treating in the dispersion for 30min, and drying under reduced pressure.
4) 50mg of CUR @ NDs-TPGS4000Redispersed in 10mL of distilled water under ultrasound for 10 min.
The resulting CUR @ NDs-TPGS4000The average particle size is 435.09nm, the particle size distribution is 0.35, and the drug loading efficiency is 30.46%.
EXAMPLE 13 preparation of CUR @ NDs-TPGS oral Nanocomposite
(1) NDs-TPGS synthesis method
1) 0.4g of NDs was added to 20mL of mixed acid (H) at 70 deg.C2SO4-HCl in a volume ratio of 3:1), stirring for 36 hours, cooling to room temperature, diluting with distilled water, and centrifuging at 10,000rpm for 15 min; heating NDs particles in 0.1M NaOH at 90 ℃ for 2 hours, and heating in 0.1M HCl at 90 ℃ for 2 hours to reduce the pH value to 5-6; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
2) 0.08g of NDs-COOH was dispersed in 20mL of oxalyl chloride by sonication, 0.2mL of dimethylformamide was added, the reaction was stirred at 70 ℃ for 24 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried in vacuum for 24 hours.
3) 8mg of NDs-COCl was ultrasonically dispersed in 10mL of dimethylformamide, followed by the addition of 60mg of TPGS2000Reacting at 100 deg.C for 36 hr, filtering the reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS2000。
(2) Preparation method of CUR @ NDs-TPGS
1) 40mg of NDs-TPGS2000Dispersed in 10mL of dimethylformamide by probe ultrasound (480w, 10 ℃)And the ultrasonic time is 45 min.
2) 2mg of CUR was dissolved in 2mL of ethanol.
3) Dropwise adding the CUR solution to NDs-TPGS under ultrasound2000Ultrasonic treating in the dispersion for 60min, and drying under reduced pressure.
4) 50mg of CUR @ NDs-TPGS2000Redispersed in 10mL of distilled water under ultrasound for 10 min.
The resulting CUR @ NDs-TPGS2000The average particle size is 164.27nm, the particle size distribution is 0.17, and the drug loading efficiency is 91.47%.
Example 14 preparation of CUR @ NDs-TPGS oral Nanocomposite
(1) NDs-TPGS synthesis method
1) 0.6g of NDs was added to 20mL of mixed acid (H) at 80 deg.C2SO4-HCl in a volume ratio of 3:1), stirring for 48 hours, cooling to room temperature, diluting with distilled water, and centrifuging at 10,000rpm for 15 min; heating NDs particles in 0.1M NaOH at 90 ℃ for 2 hours, and heating in 0.1M HCl at 90 ℃ for 2 hours to reduce the pH value to 5-6; the obtained NDs-COOH was washed with distilled water three times and vacuum-dried for 24 hours for use.
2) 0.15g of NDs-COOH was dispersed in 20mL of oxalyl chloride by sonication, 0.8mL of dimethylformamide was added, the reaction was stirred at 60 ℃ for 30 hours, NDs-COCl was washed three times with anhydrous tetrahydrofuran after centrifugation, and dried under vacuum for 24 hours.
3) 6mg of NDs-COCl was ultrasonically dispersed in 10mL of dimethylformamide, followed by addition of 40mg of TPGS1000Reacting at 100 deg.C for 40 hr, filtering the reaction solution with 0.22 μm microporous membrane, washing with water, and freeze drying to obtain NDs-TPGS1000。
(2) Preparation method of CUR @ NDs-TPGS
1) 40mg of NDs-TPGS1000Dispersed in 10mL of dimethylformamide by probe sonication (600w, 0 ℃) for 30 min.
2) 1mg of CUR was dissolved in 2mL of acetone.
3) Dropwise adding the CUR solution to NDs-TPGS under ultrasound1000Ultrasonic treating in the dispersion for 30min, and drying under reduced pressure.
4) 50mg of CUR @ NDs-TPGS2000Redispersed in 10mL of distilled water under ultrasound for 10 min.
The resulting CUR @ NDs-TPGS2000The average particle size is 141.44nm, the particle size distribution is 0.18, and the drug loading efficiency is 93.33%.
Example 15 in vivo pharmacokinetic Studies in rats
Selection of CUR @ NDs-TPGS prepared according to example 91000Selecting 0.5% sodium carboxymethylcellulose suspension of CUR and CUR @ NDs-COOH/TPGS as tested preparation1000(physical modification group) is a reference formulation. Male Wistar rats (weighing 220 + -20 g) were randomly divided into three groups of six, and fasted and freely drunk water 12h before administration. Orally administered at a dose of 75mg/kg by gavage, by taking blood from the orbit at 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 5, 6, 8 and 12 hours after administration, placing in a heparin-coated centrifuge tube, centrifuging at 10,000 Xg for 5min, separating plasma, and placing in a refrigerator at-20 ℃ for use. The plasma treatment method comprises precisely measuring 200 μ l of plasma sample, adding 50 μ l of internal standard emodin, extracting with 200 μ l of ethyl acetate, vortexing for 3min, centrifuging for 5min at 10,000 Xg, separating organic layer to another centrifuge tube, blowing with nitrogen, dissolving with 200 μ l of mobile phase, vortexing for 3min, centrifuging for 5min at 10,000 Xg, and collecting supernatant and injecting into high performance liquid chromatograph. The chromatogram and peak area were recorded and the blood concentration and the major pharmacokinetic parameters in the samples were calculated at each time point (see table 7).
TABLE 11 pharmacokinetic parameters of orally administered CUR formulations in rats
As a result, C of CUR @ NDs-TPGSmaxIs 5.71 and 1.21 times of the CUR suspension and the CUR @ NDs-COOH/TPGS group, AUC0-tIs 16.15 times and 1.37 times of the group of the CUR suspension and the CUR @ NDs-COOH/TPGS, obviously improves the oral bioavailability of the CUR, and shows good slow release effect.
Claims (10)
1. An oral curcumin-nanodiamond composite, characterized by being prepared by the following method: sequentially carrying out carboxylation and acylchlorination on the nano-diamond, and further reacting the acylchlorinated nano-diamond with vitamin E polyethylene glycol succinate to synthesize a nano-diamond-vitamin E polyethylene glycol succinate nano-drug carrier; and (3) coating curcumin in a nanocage formed by the nanodiamond-vitamin E polyethylene glycol succinate to obtain the oral curcumin-nanodiamond compound.
2. An oral curcumin-nanodiamond composite according to claim 1, wherein said nanodiamond-vitamin E polyethylene glycol succinate is synthesized by the following method:
(1) adding the nano-diamond into a certain volume of mixed acid at a certain temperature, stirring for a certain time, cooling to room temperature, washing with water and centrifuging; adjusting pH to 5-6 with sodium hydroxide and hydrochloric acid; and washing with water, and drying in vacuum to obtain the carboxyl nano-diamond.
(2) Ultrasonically dispersing carboxyl nano-diamond in oxalyl chloride, adding dimethylformamide, stirring and reacting for a certain time at a certain temperature, centrifuging, washing the carboxyl nano-diamond with anhydrous tetrahydrofuran, and drying in vacuum to obtain the acyl chloride nano-diamond.
(3) Ultrasonically dispersing acyl chloride nano-diamond in dimethylformamide, adding vitamin E polyethylene glycol succinate, reacting at a certain temperature for a certain time, washing the cooled reaction solution with water, and freeze-drying to obtain the nano-diamond-vitamin E polyethylene glycol succinate.
3. An oral curcumin-nanodiamond composite according to claim 2, wherein the mixed acid in step (1) is one of sulfuric acid-nitric acid, sulfuric acid-perchloric acid and sulfuric acid-hydrochloric acid, and the volume ratio is 5:1 to 1:2, and preferably ranges from 5:1 to 2: 1.
4. An oral curcumin-nanodiamond composite according to claim 2, wherein the concentration of the nanodiamond in the mixed acid in step (1) is 5-55 mg/mL, preferably in the range of 5-40 mg/mL; the reaction temperature of the nano-diamond in the mixed acid is 25-100 ℃.
5. An oral curcumin-nanodiamond composite according to claim 2, wherein the concentration of the carboxynanodiamond in oxalyl chloride in step (2) is 2-16 mg/mL, preferably in the range of 2-8 mg/mL.
6. An oral curcumin-nanodiamond composite according to claim 2, wherein the concentration of the acid chloride nanodiamond in dimethylformamide in step (3) is 0.1-10 mg/mL, preferably in the range of 0.1-5 mg/mL; the concentration of the vitamin E polyethylene glycol succinate in the dimethylformamide in the step (3) is 1-20 mg/mL, and the preferable range is 3-20 mg/mL.
7. An oral curcumin-nanodiamond complex according to claim 2, wherein said vitamin E polyethylene glycol succinate of step (3) is one of vitamin E polyethylene glycol 400 succinate, vitamin E polyethylene glycol 1000 succinate, vitamin E polyethylene glycol 2000 succinate, vitamin E polyethylene glycol 4000 succinate and vitamin E polyethylene glycol 6000 succinate.
8. An oral curcumin-nanodiamond composite according to claim 1, wherein said oral curcumin-nanodiamond composite is prepared by the following method:
(1) adding a certain amount of nano diamond-vitamin E polyethylene glycol succinate into a proper solvent, and ultrasonically dispersing for a certain time at a certain temperature by using a probe with certain power;
(2) dissolving a prescribed amount of curcumin in a suitable solvent;
(3) dripping the curcumin solution into the nanodiamond-vitamin E polyethylene glycol succinate suspension, continuing to perform probe ultrasonic treatment, and drying under reduced pressure to remove the organic solvent to obtain an oral curcumin-nanodiamond compound;
(4) adding distilled water into the oral curcumin-nanodiamond compound, and performing redispersion by using a probe to obtain the curcumin-nanodiamond compound.
9. An oral curcumin-nanodiamond composite according to claim 8, wherein the concentration of said nanodiamond-TPGS in step (1) is 1-30 mg/mL, preferably in the range of 5-30,
the nano-diamond-vitamin E polyethylene glycol succinate is one of nano-diamond-vitamin E polyethylene glycol 400 succinate, nano-diamond-vitamin E polyethylene glycol 1000 succinate, nano-diamond-vitamin E polyethylene glycol 2000 succinate and nano-diamond-vitamin E polyethylene glycol 4000 succinate.
10. An oral curcumin-nanodiamond complex according to claim 8, wherein the concentration of curcumin in step (2) is 0.5-3 mg/mL, preferably in the range of 0.5-2 mg/mL, and the solvent in step (2) is one of ethanol, dimethyl sulfoxide, dimethylformamide, acetone and dichloromethane.
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