Multi-target anti-cancer composition based on space-time controlled release and synergistic effect and preparation method of multi-target anti-cancer composition

Abstract

The invention discloses a multi-target anticancer composition based on space-time controlled release and synergistic effect and a preparation method thereof, the composition is composed of three oral granules with synergistic functions: immune basic granules used for establishing an immune barrier; the targeted controlled-release particles are used for targeted release of the CD47 inhibitor and the flora prodrug in the intestinal tract; the space-time programming particle is used for carrying out in-vivo epigenetic editing under light control. Through dosage form design and a multi-level controlled release technology, time sequence and space precise treatment under oral administration is achieved, the medicine has the advantages of being high in synergy, high in targeting performance, good in safety and good in patient compliance, and a brand new solution is provided for radical treatment of cancers.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a multi-target anticancer composition based on spatiotemporal controlled release and synergistic effect and a preparation method thereof. BACKGROUND

[0002] Cancer is a major global public health problem. Current treatment methods such as chemotherapy, radiotherapy and targeted therapy often face problems such as drug resistance, systemic toxicity and inability to completely remove tumor stem cells and micrometastases. Although immune checkpoint inhibitors have achieved success in some cancers, the response rate is limited and may cause immune-related adverse reactions.

[0003] There are some oral anticancer supplements in the prior art, such as compositions containing curcumin, resveratrol and other antioxidants, but their mechanisms are single, the bioavailability is low, they lack targeting, and the anticancer effect is weak. In addition, although some studies have explored the use of CRISPR and other gene editing technologies for cancer treatment, their delivery systems (usually intravenous injection) are complex, costly, and have off-target risks, making it difficult to achieve precise regulation in space and time in the body.

[0004] Therefore, there is an urgent need in the art for a system that can overcome the above-mentioned deficiencies, achieve efficient, synergistic, precise and safe systemic anticancer treatment through oral route. SUMMARY

[0005] The purpose of the present application is to provide a multi-target anticancer composition based on spatiotemporal controlled release and synergistic effect and a preparation method thereof, which solves the problems of low efficiency, high toxicity, poor targeting and inability to cure in the prior art.

[0006] The above technical purpose of the present application is achieved by the following technical solution: A multi-target anticancer composition based on spatiotemporal controlled release and synergistic effect, the composition comprising immune basic particles, targeted controlled release particles and spatiotemporal programming particles.

[0007] Further preferably, the immune basic particles comprise a fast-release portion and a slow-release portion, the fast-release portion comprising an immune modulator beta-glucan and a curcumin phospholipid complex, and the slow-release portion comprising a slow-release pellet composed of a hydroxypropyl methylcellulose skeleton, the pellet containing resveratrol and epigallocatechin gallate.

[0008] Further preferably, the targeted controlled release particles are composed of a drug-loaded pellet core and an outer coating layer, the drug-loaded pellet core comprising an immune checkpoint CD47 inhibitor, a metabolic modulator and a probiotic activation prodrug; the outer coating layer is a pH-responsive enteric material.

[0009] Further preferably, the metabolic modulator is metformin.

[0010] Further preferably, the pH-responsive enteric material is Eudragit FS30D.

[0011] Further preferably, the spatiotemporal programming particle comprises an engineered microorganism core and at least two layers of functional coating layers, the engineered microorganism core is carrying a light-controlled CRISPR-dCas9 epigenetic editing system; the coating layers at least include an enzyme-responsive layer and an enteric layer from inside to outside.

[0012] Further preferably, the coating layer further comprises a time delay layer between the enzyme-responsive layer and the enteric layer, the innermost enzyme-responsive layer is composed of a material capable of being degraded by tumor-related specific enzymes, such as guar gum; the intermediate time delay layer is composed of a hydrophilic gel material, such as hydroxypropyl methyl cellulose; and the outermost enteric coating layer is a pH-responsive material.

[0013] Further preferably, the pH-responsive material is Eudragit L100-55.

[0014] Further preferably, the genetically engineered microorganism in the engineered microorganism core is any one of lactic acid bacteria and yeast.

[0015] The application also provides a preparation method of a multi-target anticancer composition based on spatiotemporal controlled release and synergistic effect, characterized in that: S1, the preparation method of the immune-based particle comprises: preparing immediate-release particles and sustained-release particles respectively, and finally physically mixing the two, wherein the immediate-release particles are prepared by a dry granulation process; and the sustained-release particles are prepared by a wet granulation process, using hydroxypropyl methyl cellulose as a skeleton material, and obtaining after drying and granulating; S2, the preparation method of the targeted controlled-release particle comprises: preparing a drug-containing pellet core containing a CD47 inhibitor, metformin and a bacterial flora prodrug by an extrusion-spheronization method; then coating the pellet core with a pH-responsive polymer coating liquid in a fluidized bed coating device, and performing solidification treatment after coating is completed; S3, the preparation method of the spatiotemporal programming particle comprises: first, under sterile conditions, fermenting and culturing the engineered lactic acid bacteria, mixing them with trehalose and mannitol as lyophilization protectants, and preparing lyophilized pellets by drop pill and freeze-drying technology; then, in a fluidized bed coating device, sequentially spraying enzyme-responsive layer coating liquid, time delay layer coating liquid and enteric coating layer coating liquid to form a three-layer controlled-release structure, and the whole process needs to be carried out in a strictly controlled low-temperature environment.

[0016] In summary, the application has the following beneficial effects: Firstly, synergy and systematicity: This invention is the first to integrate three major functions—immune foundation building, targeted regulation, and epigenetic editing—into a single oral regimen. The timing and spatial positioning of the three granules are interconnected, forming a complete "preparation-attack-remodeling" therapeutic loop, resulting in a synergistic effect of "1+1+1>3".

[0017] Secondly, precision and safety: Through pH-responsive, enzyme-responsive, and time-controlled release technologies, precise delivery and release of drugs at the lesion site are achieved. In particular, the introduction of the light-controlled CRISPR system strictly limits gene editing activity to specific times and spaces, greatly reducing off-target risks and systemic toxicity, which is not available in existing technologies.

[0018] Thirdly, the dosage form offers advantages and improves patient compliance: All medications are in granule form, standardizing the dosage and facilitating both production and patient administration. Granules can be easily dissolved in water or mixed with food, making them particularly suitable for patients with swallowing difficulties, significantly improving treatment adherence.

[0019] Fourth, radical cure potential: This program not only targets the primary tumor, but also aims to eliminate latent micrometastases and reverse the malignant phenotype of tumor cells through systematic immune training and epigenetic editing, providing a possible technical path to achieve "functional cure" or "zeroing out" of cancer. Attached Figure Description

[0020] Figure 1 This is a schematic diagram illustrating the synergistic mechanism of the three granules in this invention; Figure 2 This is a cross-sectional view of the multi-layer structure of the spatiotemporal programming particles in this invention. Detailed Implementation

[0021] The present invention will be further described in detail below with reference to the accompanying drawings.

[0022] Example 1: This invention provides a multi-target anticancer composition based on spatiotemporal controlled release and synergistic effects. This composition consists of three separately packaged oral granules: immune-based granules, targeted controlled-release granules, and spatiotemporally programmed granules, such as... Figure 1 As shown, these three granules need to be used sequentially or in combination to produce a synergistic anti-cancer effect.

[0023] The immune foundation granule is a mixed granule composed of immediate-release and sustained-release parts. The immediate-release part contains the immunomodulator β-glucan and the high-bioavailability curcumin phospholipid complex, which is used to quickly activate innate immunity and exert anti-inflammatory effects. The sustained-release part contains sustained-release pellets composed of a hydroxypropyl methylcellulose skeleton, which encapsulates resveratrol and epigallocatechin gallate, to achieve sustained release in the intestine, maintain stable blood drug concentrations, and regulate metabolism and epigenetics. The core function of the granule is to reshape the intestinal flora and train immune cells such as macrophages, laying a solid immune foundation for subsequent therapy.

[0024] The targeted controlled-release granule is composed of a drug-loaded pellet core and an outer coating layer. The drug-loaded pellet core contains an immune checkpoint CD47 inhibitor, a metabolic regulator metformin, and a prodrug compound that can be metabolically activated by specific intestinal flora. The outer coating layer is composed of a pH-responsive polymer material, preferably Eudragit FS30D, which ensures that the granule does not release in the stomach and duodenum, but precisely dissolves and releases the contents in the neutral to weakly alkaline environment of the ileum and colon. The function of this granule is synergistic: the CD47 inhibitor removes the "don't eat me" signal of cancer cells, while the macrophages pre-trained by the immune foundation granule can more effectively perform phagocytosis; at the same time, the flora prodrug is metabolized locally into a highly active anticancer substance, achieving a "local explosion" of the tumor microenvironment; metformin enhances the sensitivity of cancer cells to treatment through metabolic remodeling.

[0025] The spatiotemporal programming granule is the most core innovation of the present application, as shown in Figure 2 The engineered microorganism core is a genetically engineered lactic acid bacterium or yeast that stably carries a light-controlled CRISPR-dCas9 gene editing system. This system can be activated under specific wavelength light to reprogram the epigenetic state of target cells, such as activating tumor suppressor genes. The three-layer coating layer includes: the innermost enzyme-responsive layer, composed of materials such as guar gum that can be degraded by tumor-associated specific enzymes; the middle time-delay layer, composed of hydrophilic gel materials such as hydroxypropyl methylcellulose, for controlled-release time; and the outermost enteric coating layer, composed of pH-responsive materials such as Eudragit L100-55. After oral administration, the granule releases engineered microorganisms at a predetermined time sequence at a specific site in the intestine, which are taken up by immune cells and carried to the whole body. At the predetermined time point, the gene editing function is remotely and accurately activated by in vitro non-invasive specific wavelength light, thereby "resetting" the circulating tumor cells or micrometastases at the epigenetic level.

[0026] Example II, a method for preparing a multi-target anticancer composition based on spatiotemporal controlled release and synergistic action, comprising the following steps: S1, Preparation of immune-based granules Take 2000 grams of yeast beta-glucan, 500 grams of curcumin phospholipid complex, and 475 grams of microcrystalline cellulose, and mix them in a three-dimensional mixer for 15 minutes. Use a dry granulation machine to compress, crush and granulate to obtain immediate-release granules. Take 150 grams of resveratrol-cyclodextrin inclusion compound, 100 grams of EGCG, 750 grams of hydroxypropyl methylcellulose K100M, and anhydrous lactose to 1500 grams, use purified water as the binder, and pass through a high-speed shearing granulator, fluidized bed drying and granulation to obtain sustained-release granules. Mix the immediate-release granules and sustained-release granules in a ratio of 7:3 by weight into a double-cone mixer and mix for 20 minutes. Use aluminum-plastic composite film to package, 5 grams per bag.

[0027] S2, Preparation of targeted controlled-release granules Take 50 grams of CD47 inhibitory polypeptide, 500 grams of metformin hydrochloride, 100 grams of MDA-01, and microcrystalline cellulose to 1000 grams, mix them, and then pass them through an extrusion-spheronization machine to form pellet cores with a particle size of 0.8 to 1.2 millimeters. Prepare a coating liquid composed of Eudragit FS30D 300 grams, triethyl citrate 60 grams, and talc 90 grams. Coat the pellet cores in a fluidized bed bottom spraying device, and control the coating weight gain at 25%. After coating, solidify at 40 degrees Celsius for 2 hours, and package, 3 grams per bag.

[0028] S3, Preparation of space-time programming granules In the A-level clean area under the B-level background, cultivate the engineered lactic acid bacteria to the logarithmic growth phase, and centrifuge to harvest the bacterial cells. Mix the wet bacterial cells with a freeze-drying protection solution containing 800 grams of trehalose and 500 grams of mannitol. Use a precision pellet dropping device to drop the bacterial suspension into liquid nitrogen for instant molding, then transfer it to a freeze dryer for main drying and analytical drying to obtain live bacteria freeze-dried pellets. In the fluidized bed coating equipment, spray guar gum solution to form an enzyme-responsive layer, then spray hydroxypropyl methylcellulose E5 solution to form a time-delay layer, and then spray Eudragit L100-55 solution to form an enteric layer. Finally, the final product is packaged in an aluminum foil bag under low temperature conditions and dry agent is added.

[0029] This specific embodiment is only an explanation of the present application, which is not a limitation of the present application. Those skilled in the art can make modifications to this embodiment without creative contribution after reading this specification, but as long as it is within the scope of the claims of the present application, it is protected by the patent law.

Claims

1. A multi-target anticancer composition based on spatiotemporal controlled release and synergism, characterized in that: The composition comprises immune base particles, targeted controlled release particles and space-time programming particles.

2. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 1, characterized in that: The immune base particles comprise a fast release part and a slow release part, the fast release part contains an immune modulator β-glucan and a curcumin phospholipid complex, and the slow release part contains a slow release pellet composed of a hydroxypropyl methyl cellulose skeleton, which encapsulates resveratrol and epigallocatechin gallate.

3. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 2, characterized in that: The targeted controlled release particles are composed of a drug-loaded pellet core and an outer coating layer, the drug-loaded pellet core contains an immune checkpoint CD47 inhibitor, a metabolic regulator and a flora activating prodrug; the outer coating layer is a pH-responsive enteric material.

4. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 3, characterized in that: The metabolic regulator is metformin.

5. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 4, characterized in that: The pH-responsive enteric material is Eudragit FS30D.

6. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 5, characterized in that: The space-time programming particles comprise an engineered microorganism core and at least two layers of functional coating layers, the engineered microorganism core is a light-controlled CRISPR-dCas9 epigenetic editing system; the coating layers at least include an enzyme-responsive layer and an enteric layer from inside to outside.

7. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 6, characterized in that: The coating layer also includes a time delay layer between the enzyme-responsive layer and the enteric layer, the innermost enzyme-responsive layer is composed of a material that can be degraded by tumor-related specific enzymes, such as guar gum; the middle time delay layer is composed of a hydrophilic gel material, such as hydroxypropyl methyl cellulose; the outermost enteric coating layer is a pH-responsive material.

8. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 7, characterized in that: The pH-responsive material is Eudragit L100-55.

9. The multi-target anticancer composition based on spatiotemporal controlled release and synergy according to claim 8, characterized in that: The genetically engineered microorganism in the engineered microorganism core is any one of lactic acid bacteria and yeast.

10. The preparation method of the multi-target anti-cancer composition based on space-time controlled release and synergy according to claim 9, characterized in that: S1, the preparation method of the immune base particles comprises: preparing fast release particles and slow release particles respectively, and finally physically mixing the two, wherein the fast release particles are prepared by dry granulation process; the slow release particles are prepared by wet granulation process, using hydroxypropyl methyl cellulose as the skeleton material, and obtaining after drying and granulating; S2, the preparation method of the targeted controlled release particles comprises: preparing a drug-loaded pellet core containing a CD47 inhibitor, metformin and a flora prodrug by extrusion and spheronization method; then using a pH-responsive polymer coating liquid to coat the pellet core in a fluidized bed coating device, and performing solidification treatment after coating is completed; S3, the preparation method of the space-time programming particles comprises: first, under sterile conditions, fermenting and culturing the engineered lactic acid bacteria, mixing them with the freeze-drying protectant trehalose and mannitol, and preparing freeze-dried pellets by drop pill and freeze-drying technology; then, in a fluidized bed coating device, sequentially spraying enzyme-responsive layer coating liquid, time delay layer coating liquid and enteric coating layer coating liquid to form a three-layer controlled release structure, and the whole process needs to be carried out in a strictly controlled low-temperature environment.

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