A cordycepin-v type starch inclusion compound and a preparation method thereof

Abstract

The application discloses a cordycepin-V type starch inclusion compound and a preparation method thereof, wherein the cordycepin is used as a guest to be included, and the V type starch is used as a carrier, and the relative crystallinity of the V type starch is 11.07-25.12%. In the preparation method, the V type starch is prepared by using a glycerol ethanol hot substitution method, and then the cordycepin-V type starch inclusion compound is prepared by weighing the cordycepin and the V type starch according to a mass ratio of 1:80. The prepared inclusion compound can improve the stability of the cordycepin in gastric juice and prolong the intestinal release time of the cordycepin under the premise of ensuring the loading amount of the cordycepin, and provides a new thought and theoretical guidance for effectively improving the bioavailability of the cordycepin.

Description

Technical Field

[0001] This invention relates to the field of cordycepin inclusion complexes, specifically a cordycepin-V type starch inclusion complex and its preparation method. Background Technology

[0002] Cordycepin (3'-deoxyadenosine), mainly produced from Cordyceps militaris, belongs to the purine alkaloid class and is a novel nucleoside antibiotic with anti-inflammatory, antiviral, antitumor, and immunomodulatory effects. It can inhibit the release of inflammatory factors such as TNF-α and IL-1β, reducing inflammatory responses; it can interfere with nucleic acid metabolism in tumor cells, block the G1 / S phase transition of the cell cycle, and induce apoptosis. Therefore, developing cordycepin as a widely applicable new drug or health food is an inevitable trend.

[0003] However, cordycepin's bioavailability is significantly reduced due to the acidic environment and enzymatic hydrolysis of the gastrointestinal tract. The highly acidic environment of human gastric juice catalyzes the breakage of the nucleoside glycosidic bond (N-glycosidic bond) in the cordycepin molecule, causing it to lose its biological activity. Furthermore, adenosine deaminase and nucleoside phosphorylase present on the surface of intestinal epithelial cells and in the intestinal lumen specifically degrade cordycepin, resulting in a very short effective retention time in the intestinal lumen. Therefore, after direct ingestion of cordycepin by the human body, only a very small amount of active cordycepin can successfully reach the intestinal epithelial cells and enter the bloodstream for absorption and utilization.

[0004] Currently, the inclusion technology for cordycepin is not yet mature. This is mainly because cordycepin, as a bioactive ingredient highly sensitive to light, heat, and acid, has high requirements for inclusion materials. Current inclusion materials for cordycepin primarily focus on cyclodextrin, but this involves the need for frequent, high-dose administration, leading to side effects, and also presents high costs, severely limiting the application range of cordycepin. Summary of the Invention

[0005] This invention provides a cordycepin-V type starch inclusion complex and its preparation method, in order to solve the problems of easy side effects and high cost of cordycepin inclusion complexes in the prior art.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A cordycepin-V-type starch inclusion complex, wherein cordycepin is the inclusion object and V-type starch is the carrier, with a loading content of 23.39-34.56 mg / g, wherein the relative crystallinity of V-type starch is 11.07-25.12%.

[0007] Furthermore, the relative crystallinity of the V-type starch is 11.07%, and the corresponding loading content is 23.39 mg / g.

[0008] Furthermore, the relative crystallinity of the V-type starch is 14.20%, and the corresponding loading content is 28.67 mg / g.

[0009] Furthermore, the relative crystallinity of the V-type starch is 25.12%, and the corresponding loading content is 34.56 mg / g.

[0010] A method for preparing the above-mentioned cordycepin-V type starch inclusion complex is as follows: Step 1: Using corn starch as raw material, V-type starch with a relative crystallinity of 11.07-25.12% was prepared by the glycerol-ethanol thermal substitution method. Step 2: Weigh cordycepin and the V-type starch at a mass ratio of 1:80 to prepare a cordycepin-V-type starch inclusion complex with a loading content of 23.39-34.56 mg / g.

[0011] Furthermore, in step 1, the process of preparing V-type starch by the glycerol-ethanol thermal substitution method is as follows: Prepare a 5-10% corn starch glycerol solution, and heat the corn starch glycerol solution at 140-145 ℃ with continuous stirring for 20-40 min to obtain a gelatinized corn starch solution. Then cool the gelatinized corn starch solution to 90-95 ℃. A 40%-100% ethanol aqueous solution is preheated at 90-95℃ for 10-15 minutes. Then, a gelatinized corn starch solution cooled to 90-95℃ is added to a preheated ethanol aqueous solution with a volume four times its own volume. The mixture is then heated at 90-95℃ for 1-2 hours to obtain a mixture. The mixture was cooled to room temperature, then centrifuged to remove the supernatant and washed to obtain a precipitate. The precipitate was then dried at 40-45℃ for 12-24 hours to obtain V-type starch with a relative crystallinity of 11.07-25.12%.

[0012] Furthermore, the concentration of the ethanol-water solution is 40%, thereby obtaining V-type starch with a relative crystallinity of 11.07%.

[0013] Furthermore, the concentration of the ethanol aqueous solution is 60%, thereby obtaining V-type starch with a relative crystallinity of 25.12%.

[0014] Furthermore, the concentration of the ethanol aqueous solution is 100%, thereby obtaining V-type starch with a relative crystallinity of 14.20%.

[0015] Further, in step 2, the weighed cordycepin and the V-type starch are prepared into a suspension using a 60% ethanol aqueous solution; the suspension is then stirred at a constant temperature of 40-50°C in the dark for 3-4 hours and then rapidly cooled to 4°C to obtain a mixture; the mixture is centrifuged to remove the supernatant and then washed to obtain a precipitate, which is then freeze-dried at -80°C for 48 hours to obtain a cordycepin-V-type starch inclusion complex with a loading content of 23.39-34.56 mg / g.

[0016] In this invention, corn starch is used as raw material. The corn starch is induced to form a V-shaped crystalline structure starch with a relative crystallinity in the range of 11.07-25.12% by the glycerol-ethanol thermal substitution method. The spiral cavity formed by the V-shaped starch is used to achieve the inclusion effect of cordycepin, thereby obtaining a cordycepin-V-shaped starch inclusion complex with enhanced cordycepin stability.

[0017] In the glycerol-ethanol thermal substitution method, the relative crystallinity of V-type starch induced by different concentrations of ethanol-water solutions varies. Specifically, the V-type starch induced by a 40% concentration of ethanol-water solution has a relative crystallinity of 11.07%, with a corresponding cordycepin-V-type starch inclusion complex loading of 23.39 mg / g. The V-type starch induced by a 60% concentration of ethanol-water solution has a relative crystallinity of 25.12%, with a corresponding cordycepin-V-type starch inclusion complex loading of 34.56 mg / g; and the V-type starch induced by a 100% concentration of ethanol-water solution has a relative crystallinity of 14.20%, with a corresponding cordycepin-V-type starch inclusion complex loading of 28.67 mg / g.

[0018] Therefore, the cordycepin-V type starch inclusion complex obtained by the present invention can produce V type starch with a relative crystallinity in the range of 11.07-25.12%, and thus cordycepin-V type starch inclusion complexes with different release capacities can be prepared as needed.

[0019] This invention utilizes V-shaped starch prepared by a glycerol-ethanol thermal substitution method as an encapsulation material, achieving a cordycepin loading capacity of 23.39-34.56 mg / g. By loading inclusion complexes with varying cordycepin content, the stability of cordycepin in the human gastrointestinal tract is effectively enhanced, its release in a simulated gastrointestinal tract is controlled, its bioavailability is improved, and the side effects caused by frequent high-dose administration are avoided.

[0020] In this invention, cordycepin-V-type starch inclusion complexes, formed by loading cordycepin with V-type starch of varying relative crystallinity, exhibit reconstructed internal hydrogen bond networks and enhanced stability. Simultaneously, they control the release of cordycepin in simulated gastrointestinal digestive fluids, delaying the intestinal release time and facilitating its absorption. The controlled release capability of the cordycepin-V-type starch inclusion complexes formed from V-type starch is related to intermolecular forces and stability.

[0021] The inclusion complex prepared by this invention improves the stability of cordycepin in gastric juice and prolongs its intestinal release time while ensuring the loading of cordycepin, providing new ideas and theoretical guidance for effectively improving the bioavailability of cordycepin. Attached Figure Description

[0022] Figure 1 This is a flowchart of the preparation method according to an embodiment of the present invention.

[0023] Figure 2A The XRD diffraction patterns are those of ordinary corn starch NCS and V-type starches (V-40%, V-60%, and V-100%) prepared from 40%, 60%, and 100% ethanol aqueous solutions in Examples 1, 2, and 3, respectively.

[0024] Figure 2B This is the XRD diffraction pattern of cordycepin (Cor).

[0025] Figure 3A These are Fourier transform infrared (FTIR) spectra of V-type starch (V-40%, V-60%, and V-100%) prepared with 40%, 60%, and 100% ethanol concentrations in Examples 1, 2, and 3, respectively.

[0026] Figure 3B The Fourier transform infrared (FTIR) spectra of cordycepin (Cor), cordycepin-gelatinized corn starch complex (GS-Cor), and the 40%-Cor, 60%-Cor, and 100%-Cor cordycepin-V type starch inclusion complexes prepared in Examples 1, 2, and 3.

[0027] Figure 4A The contact angles of V-type starches prepared with 40%, 60%, and 100% ethanol in Examples 1, 2, and 3 are V-40%, V-60%, and V-100%, respectively.

[0028] Figure 4B The contact angles are those of the cordycepin-gelatinized corn starch complex (GS-Cor) and the cordycepin-V-type starch inclusion complexes 40%-Cor, 60%-Cor, and 100%-Cor prepared in Examples 1, 2, and 3.

[0029] Figure 5AThese are the TG thermogravimetric curves of cordycepin (Cor), cordycepin-gelatinized corn starch complex (GS-Cor), and the 40%-Cor, 60%-Cor, and 100%-Cor cordycepin-V type starch inclusion complexes prepared in Examples 1, 2, and 3.

[0030] Figure 5B The images show the DTG thermogravimetric curves of cordycepin (Cor), cordycepin-gelatinized corn starch complex (GS-Cor), and the 40%-Cor, 60%-Cor, and 100%-Cor cordycepin-V type starch inclusion complexes prepared in Examples 1, 2, and 3.

[0031] Figure 6A This is a graph showing the loading content of cordycepin-gelatinized corn starch complex (GS-Cor) and cordycepin-V type starch inclusion complexes 40%-Cor, 60%-Cor, and 100%-Cor prepared in Examples 1, 2, and 3.

[0032] Figure 6B The release rates of cordycepin (Cor), cordycepin-gelatinized corn starch complex (GS-Cor), and the 40%-Cor, 60%-Cor, and 100%-Cor of cordycepin-V type starch inclusion complexes prepared in Examples 1, 2, and 3 in simulated gastrointestinal digestive fluids are described. Detailed Implementation

[0033] To enable those skilled in the art to better understand the present invention, the embodiments will be described in detail below with reference to the accompanying drawings and examples. This will allow for a full understanding of how the present invention uses technical means to solve technical problems and achieve corresponding technical effects, and to facilitate its implementation. The embodiments of the present invention and the various features within them can be combined with each other without conflict, and all resulting technical solutions are within the protection scope of the present invention.

[0034] Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort should fall within the scope of protection of the present invention.

[0035] It should be noted that the terms "comprising" and "having," and any variations thereof, in the specification, claims, and accompanying drawings of this invention are intended to cover non-exclusive inclusion. Example 1

[0036] like Figure 1 As shown in the figure, this embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0037] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140 °C for 30 min to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90 °C.

[0038] A 40% ethanol aqueous solution was preheated at 90 °C for 10 min. Then, a gelatinized corn starch solution cooled to 90 °C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90 °C for 1 h to obtain a mixture.

[0039] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40 °C for 12 h to obtain V-type starch with a relative crystallinity of 11.07%, denoted as V-40.

[0040] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0041] The suspension was then stirred at 40°C in the dark for 4 hours, followed by rapid cooling to 4°C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80°C for 48 hours to obtain a cordycepin-V starch inclusion complex with a loading concentration of 23.39 mg / g.

[0042] Therefore, in this embodiment, the cordycepin-V-type starch inclusion complex has a loading content of 23.39 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 11.07%. The cordycepin-V-type starch inclusion complex obtained in this embodiment is denoted as 40%-Cor. Example 2

[0043] This embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0044] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140 °C for 30 min to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90 °C.

[0045] A 60% ethanol aqueous solution was preheated at 90 °C for 10 min. Then, a gelatinized corn starch solution cooled to 90 °C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90 °C for 1 h to obtain a mixture.

[0046] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40 °C for 12 h to obtain V-type starch with a relative crystallinity of 25.12%, denoted as V-60%.

[0047] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0048] The suspension was then stirred at 40 °C in the dark for 4 h, followed by rapid cooling to 4 °C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80 °C for 48 h to obtain a cordycepin-V starch inclusion complex with a loading concentration of 34.56 mg / g.

[0049] Therefore, in this embodiment, the cordycepin-V-type starch inclusion complex has a loading content of 34.56 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 25.12%. The cordycepin-V-type starch inclusion complex obtained in this embodiment is denoted as 60%-Cor. Example 3

[0050] This embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0051] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140 °C for 30 min to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90 °C.

[0052] A 100% ethanol aqueous solution was preheated at 90 °C for 10 min. Then, a gelatinized corn starch solution cooled to 90 °C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90 °C for 1 h to obtain a mixture.

[0053] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40 °C for 12 h to obtain V-type starch with a relative crystallinity of 14.20%, denoted as V-100.

[0054] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0055] The suspension was then stirred at 40 °C in the dark for 4 h, followed by rapid cooling to 4 °C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80 °C for 48 h to obtain a cordycepin-V starch inclusion complex with a loading concentration of 28.67 mg / g.

[0056] Therefore, in this embodiment, the cordycepin-V-type starch inclusion complex has a loading content of 28.67 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 14.20%. The cordycepin-V-type starch inclusion complex obtained in this embodiment is denoted as 100%-Cor. Example 4

[0057] This embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0058] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140 °C for 30 min to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90 °C.

[0059] A 50% ethanol aqueous solution was preheated at 90 °C for 10 min. Then, a gelatinized corn starch solution cooled to 90 °C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90 °C for 1 h to obtain a mixture.

[0060] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40 °C for 12 h to obtain V-type starch with a relative crystallinity of 19.87%.

[0061] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0062] The suspension was then stirred at 40 °C in the dark for 4 h, followed by rapid cooling to 4 °C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80 °C for 48 h to obtain a cordycepin-V starch inclusion complex with a loading concentration of 30.98 mg / g.

[0063] Therefore, in this embodiment, the loading content of the cordycepin-V-type starch inclusion complex is 30.98 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 19.87%. Example 5

[0064] This embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0065] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140°C for 30 minutes to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90°C.

[0066] A 70% ethanol aqueous solution was preheated at 90°C for 10 minutes. Then, a gelatinized corn starch solution cooled to 90°C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90°C for 1 hour to obtain a mixture.

[0067] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40°C for 12 hours to obtain V-type starch with a relative crystallinity of 19.60%.

[0068] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0069] The suspension was then stirred at 40°C in the dark for 4 hours, followed by rapid cooling to 4°C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80°C for 48 hours to obtain a cordycepin-V starch inclusion complex with a loading concentration of 30.24 mg / g.

[0070] Therefore, in this embodiment, the loading content of the cordycepin-V-type starch inclusion complex is 30.24 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 19.60%. Example 6

[0071] This embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0072] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140°C for 30 minutes to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90°C.

[0073] An 80% ethanol aqueous solution was preheated at 90°C for 10 minutes. Then, a gelatinized corn starch solution cooled to 90°C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90°C for 1 hour to obtain a mixture.

[0074] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40°C for 12 hours to obtain V-type starch with a relative crystallinity of 16.16%.

[0075] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0076] The suspension was then stirred at 40°C in the dark for 4 hours, followed by rapid cooling to 4°C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80°C for 48 hours to obtain a cordycepin-V starch inclusion complex with a loading concentration of 29.76 mg / g.

[0077] Therefore, in this embodiment, the loading content of the cordycepin-V-type starch inclusion complex is 29.76 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 16.16%. Example 7

[0078] This embodiment discloses a method for preparing cordycepin-V-type starch inclusion complex, the process of which is as follows: Step 1: Using corn starch (NCS) with A-type crystalline structure as raw material, V-type starch with V-type crystalline structure is prepared by glycerol-ethanol thermal substitution method.

[0079] Specifically, corn starch and glycerol are prepared into a 6% corn starch-glycerol solution. The corn starch-glycerol solution is continuously stirred and heated in a constant temperature stirrer at 140°C for 30 minutes to obtain a gelatinized corn starch solution. Then, the gelatinized corn starch solution is cooled to 90°C.

[0080] A 90% ethanol aqueous solution was preheated at 90°C for 10 minutes. Then, a gelatinized corn starch solution cooled to 90°C was added to a preheated ethanol aqueous solution with a volume four times that of the gelatinized corn starch solution. The mixture was then heated at 90°C for 1 hour to obtain a mixture.

[0081] The mixture was then cooled to room temperature, centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. The precipitate was then dried at 40°C for 12 hours to obtain V-type starch with a relative crystallinity of 15.02%.

[0082] Step 2: Weigh 5 mg of cordycepin (Cor) and 400 mg of the V-type starch obtained in Step 1 at a mass ratio of 1:80. Prepare a suspension of the weighed cordycepin and V-type starch using a 60% ethanol aqueous solution.

[0083] The suspension was then stirred at 40°C in the dark for 4 hours, followed by rapid cooling to 4°C to obtain a mixture. The mixture was then centrifuged to remove the supernatant, and washed three times with anhydrous ethanol to obtain a precipitate. Finally, the precipitate was freeze-dried at -80°C for 48 hours to obtain a cordycepin-V starch inclusion complex with a loading concentration of 29.05 mg / g.

[0084] Therefore, in this embodiment, the loading content of the cordycepin-V-type starch inclusion complex is 29.05 mg / g, wherein cordycepin is the included guest, V-type starch is the carrier, and the relative crystallinity of V-type starch is 15.02%. Contrast

[0085] Weigh out corn starch and prepare a 10% corn starch aqueous solution. Then, heat the corn starch aqueous solution in a constant temperature stirrer at 95 °C for 30 min to obtain a gelatinized corn starch solution. Subsequently, freeze-dry the solution to obtain gelatinized corn starch.

[0086] Weigh 5 mg of cordycepin and 400 mg of gelatinized corn starch, and prepare a suspension by mixing the weighed cordycepin and gelatinized corn starch with 60% ethanol aqueous solution. Then, stir at 40 °C in the dark for 4 h and then quickly cool to 4 °C to obtain a mixture.

[0087] The mixture was centrifuged and the precipitate was washed three times with anhydrous ethanol to obtain the precipitate. The obtained precipitate was freeze-dried at -80 °C for 48 h to obtain the cordycepin-gelatinized corn starch complex, denoted as GS-Cor. Experimental results

[0088] The following experimental results compare the performance of 40%-Cor obtained in Example 1, 60%-Cor obtained in Example 2, 100%-Cor obtained in Example 3, and GS-Cor obtained in the comparative example.

[0089] The V-type starch inclusion complexes (i.e., 40%-Cor, 60%-Cor, and 100%-Cor in Examples 1, 2, and 3) provided by this invention for improving the gastrointestinal stability of cordycepin are obtained by modifying ordinary corn starch (NCS) with A-type crystal characteristic diffraction peaks (15.2°, 17.1°, 18.0°, 23.1°) into V-type starch with V-type crystal characteristic diffraction peaks (7.9°, 13.7°, 21.1°), and having different relative crystallinity. Figure 2A As shown, the relative crystallinity of V-type starches V-40%, V-60%, and V-100% prepared from the 40% ethanol aqueous solution in Example 1, the 60% ethanol aqueous solution in Example 2, and the 100% ethanol aqueous solution in Example 3 are 11.07%, 25.12%, and 14.20%, respectively. Figure 2B As shown, the positions of the characteristic diffraction peaks of the unrecombined cordycepin (Cor) crystal structure are at 9.5°, 11.6°, 15.6°, 19.0°, and 26.4°, which are significantly different from the positions of the characteristic peaks of the V-type starch prepared in Examples 1, 2, and 3.

[0090] The V-type starch inclusion complexes (i.e., 40%-Cor, 60%-Cor, and 100%-Cor in Examples 1, 2, and 3) provided by this invention for improving the gastrointestinal stability of cordycepin show the following FTIR spectra: Figure 3B As shown. Figure 3A As shown, the tensile vibration of the -OH group in V-type starch (V-40%, V-60%, V-100%) occurs at 3416 cm⁻¹. -1 Nearby, while the unreacted cordycepin (Cor) was at 3330.48, 2923.51, 1667.46, and 1606.02 cm⁻¹. -1 There are distinct characteristic peaks at this point, including one at 1606.02 cm⁻¹. -1 The characteristic peaks at these locations are the signature fingerprint peaks of the cordycepin molecule, corresponding to the vibrations of its purine ring aromatic skeleton. The complexes (40%-Cor, 60%-Cor, 100%-Cor) show peaks in the range of 2961.01–2965.80 cm⁻¹. -1 The characteristic peaks appearing at 3400 and 1640 cm⁻¹ indicate the presence of cordycepin in the system. Simultaneously, the peaks at 3400 and 1640 cm⁻¹... -1 The shift of nearby characteristic peaks further illustrates that the introduction of cordycepin leads to the reconstruction of the hydrogen bond network inside the sample.

[0091] The V-type starch inclusion complexes (i.e., 40%-Cor, 60%-Cor, and 100%-Cor in Examples 1, 2, and 3) provided by this invention for improving the gastrointestinal stability of cordycepin show the following contact angle test spectra: Figure 4B As shown. Figure 4A As shown, the contact angles of V-40%, V-60%, and V-100% are all less than 90°, indicating hydrophilicity. This effectively avoids the phenomenon of hydrophobic materials being easily adsorbed by proteins in the intestine. Among them, the V-60% sample has the weakest hydrophilicity with a contact angle of 40.39°, which is because this sample has the most hydrophobic helical cavity structures. Meanwhile, after cordycepin is combined with V-type starch, the 60%-Cor complex has the smallest contact angle of 43.40°, exhibiting the strongest hydrophilicity. This is because more of the cordycepin hydrophobic structures can be fixed in the helical cavities, exposing a certain amount of hydrophilic structure, effectively inhibiting the decrease in surface hydrophilicity caused by the dehydration of the complex by anhydrous ethanol.

[0092] The V-type starch inclusion complexes (i.e., 40%-Cor, 60%-Cor, and 100%-Cor in Examples 1, 2, and 3) provided by this invention for improving the gastrointestinal stability of cordycepin, have TGA spectra as shown below. Figure 5A and 5B As shown, the maximum weight loss temperature of unreinforced cordycepin was 283.24 °C, with a weight loss rate of 1.99148% / °C. After recombination, the maximum weight loss temperatures of the 40%-Cor, 60%-Cor, and 100%-Cor complexes increased to 303.06–311.90 °C, indicating enhanced resistance to decomposition of cordycepin. Furthermore, compared to the cordycepin-gelatinized corn starch complex (GS-Cor), the cordycepin-V-type starch inclusion complexes (40%-Cor, 60%-Cor, 100%-Cor) exhibited higher maximum weight loss temperatures (305.34–311.90 °C) and slower weight loss rates (2.91058–3.15260% / °C), suggesting that V-type starch is superior to ordinary starch in improving the stability of cordycepin. Among them, the 60%-Cor sample exhibited the best stability with a maximum weight loss temperature of 311.90 ℃ and a weight loss rate of 2.91058% / ℃.

[0093] The V-type starch inclusion complexes (i.e., 40%-Cor, 60%-Cor, and 100%-Cor in Examples 1, 2, and 3) provided by this invention for improving the gastrointestinal stability of cordycepin exhibit the following effects on the loading content of cordycepin and its release effect in simulated gastrointestinal digestive fluids: Figure 6A , Figure 6B The results showed that the GS-Cor complex had a cordycepin loading of only 8.74 mg / g, while the 40%-Cor, 60%-Cor, and 100%-Cor complexes had cordycepin loadings of 23.39-34.56 mg / g. This indicates that V-type starch has an advantage over gelatinized corn starch in cordycepin loading, because the pre-formed helical cavities in V-type starch provide space for cordycepin attachment. Furthermore, according to... Figure 6BThe results showed that unreinforced cordycepin was completely released within 120 min and could not remain stable in simulated gastric juice. In contrast, the incorporation complexes (40%-Cor, 60%-Cor, 100%-Cor) exhibited a release rate of no more than 25% within 120 min, demonstrating significantly enhanced stability in simulated gastric juice. Compared to the cordycepin-gelatinized corn starch complex (GS-Cor), the 40%-Cor, 60%-Cor, and 100%-Cor complexes effectively controlled the release of cordycepin in simulated gastrointestinal juice, primarily by maintaining the stability of the cordycepin structure in simulated gastric juice, ensuring a release rate of no more than 25% during this stage, while achieving a release rate exceeding 90% in simulated intestinal juice. Furthermore, encapsulating cordycepin in a V-shaped starch structure can regulate its release kinetics and provide a certain degree of anti-degradation protection, allowing cordycepin to be transported into the small intestine in a more complete form, thus improving its bioavailability.

[0094] In summary, the 40%-Cor, 60%-Cor, and 100%-Cor prepared in Examples 1, 2, and 3 were rapidly released in simulated intestinal fluid after 120 min, and the release rate of cordycepin reached 90.30-91.20% after 300 min, which effectively improved the stability of cordycepin and prolonged its release in the gastrointestinal tract.

[0095] The preferred embodiments of the present invention have been described in detail above with reference to the accompanying drawings. These embodiments are merely descriptions of preferred embodiments and are not intended to limit the scope or concept of the invention. The specific technical features described in the above embodiments can be combined in any suitable manner without contradiction. Such combinations, as long as they do not violate the spirit of the present invention, should also be considered as part of this disclosure. To avoid unnecessary repetition, the present invention will not further describe the various possible combinations.

[0096] This invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of this invention and without departing from the design idea of ​​this invention, all modifications and improvements made by those skilled in the art to the technical solutions of this invention should fall within the protection scope of this invention. The technical content for which protection is sought in this invention has been fully described in the claims.

Claims

1. A cordycepin-V-type starch inclusion complex, characterized in that, The cordycepin is used as the encapsulated object, and V-type starch is used as the carrier, with a loading content of 23.39-34.56 mg / g, wherein the relative crystallinity of V-type starch is 11.07-25.12%.

2. The cordycepin-V type starch inclusion complex according to claim 1, characterized in that, The relative crystallinity of the V-type starch is 11.07%, and the corresponding loading content is 23.39 mg / g.

3. The cordycepin-V type starch inclusion complex according to claim 1, characterized in that, The relative crystallinity of the V-type starch is 14.20%, and the corresponding loading content is 28.67 mg / g.

4. The cordycepin-V type starch inclusion complex according to claim 1, characterized in that, The relative crystallinity of the V-type starch is 25.12%, and the corresponding loading content is 34.56 mg / g.

5. A method for preparing the cordycepin-V type starch inclusion complex as described in any one of claims 1-4, characterized in that, The process is as follows: Step 1: Using corn starch as raw material, V-type starch with a relative crystallinity of 11.07-25.12% was prepared by the glycerol-ethanol thermal substitution method. Step 2: Weigh cordycepin and the V-type starch at a mass ratio of 1:80 to prepare a cordycepin-V-type starch inclusion complex with a loading content of 23.39-34.56 mg / g.

6. The method for preparing a cordycepin-V type starch inclusion complex according to claim 5, characterized in that, In step 1, the process of preparing V-type starch by the glycerol-ethanol thermal substitution method is as follows: Prepare a 5-10% corn starch glycerol solution, and heat the corn starch glycerol solution at 140-145 ℃ with continuous stirring for 20-40 min to obtain a gelatinized corn starch solution. Then cool the gelatinized corn starch solution to 90-95 ℃. A 40%-100% ethanol aqueous solution is preheated at 90-95℃ for 10-15 minutes. Then, a gelatinized corn starch solution cooled to 90-95℃ is added to a preheated ethanol aqueous solution with a volume four times its own volume. The mixture is then heated at 90-95℃ for 1-2 hours to obtain a mixture. The mixture was cooled to room temperature, then centrifuged to remove the supernatant and washed to obtain a precipitate. The precipitate was then dried at 40-45℃ for 12-24 hours to obtain V-type starch with a relative crystallinity of 11.07-25.12%.

7. The method for preparing a cordycepin-V type starch inclusion complex according to claim 6, characterized in that, The concentration of the ethanol-water solution was 40%, thereby obtaining V-type starch with a relative crystallinity of 11.07%.

8. The method for preparing a cordycepin-V type starch inclusion complex according to claim 6, characterized in that, The concentration of the ethanol-water solution was 60%, thereby obtaining V-type starch with a relative crystallinity of 25.12%.

9. The method for preparing a cordycepin-V type starch inclusion complex according to claim 6, characterized in that, The concentration of the ethanol-water solution was 100%, thereby obtaining V-type starch with a relative crystallinity of 14.20%.

10. The method for preparing a cordycepin-V type starch inclusion complex according to claim 5, characterized in that, In step 2, the weighed cordycepin and the V-type starch were prepared into a suspension using a 60% ethanol aqueous solution. The suspension was then stirred at a constant temperature of 40-50°C in the dark for 3-4 hours, and then rapidly cooled to 4°C to obtain a mixture. The mixture was centrifuged to remove the supernatant, and then washed to obtain a precipitate. The precipitate was then freeze-dried at -80°C for 48 hours to obtain a cordycepin-V-type starch inclusion complex with a loading content of 23.39-34.56 mg / g.

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