Curcumin nanocrystal composition as well as preparation method and application thereof

Abstract

The invention relates to a curcumin nanocrystal composition, the curcumin nanocrystal composition is prepared from a curcumin nanocrystal suspension and a piperine suspension, the mass ratio of curcumin to piperine is (5-15): 1, the curcumin nanocrystal suspension contains 20-35% of curcumin nanocrystals, 1-10% of a stereo protective agent and 1-10% of a charge protective agent, the mass ratio of the stereo protective agent to the charge protective agent is (5-15): 1, and the mass ratio of the piperine to the curcumin nanocrystal suspension is (5-15): 1. Wherein the average particle size of the curcumin nanocrystal is 200 to 320 nm. The curcumin nanocrystal composition prepared by the invention can effectively prevent and treat heat stroke and complications thereof, remarkably improve heat tolerance, prolong survival time, reduce the level of inflammatory factors in blood, protect intestinal epithelial cells and intestinal barrier functions, reduce damage caused by heat stroke, prevent and treat body and organ damage caused by heat stroke, and improve the curative effect of the curcumin nanocrystal composition. The preparation has the advantages of being high in drug loading capacity, good in stability, high in bioavailability, safe, effective, convenient to carry and the like.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a curcumin nanocrystal composition, a preparation method thereof and application thereof. BACKGROUND

[0002] Heatstroke is the most severe type of heatstroke, characterized by a rapid rise in core body temperature (usually above 40℃) and central nervous system dysfunction. The pathogenesis of heatstroke is complex: first, dysfunction of the body temperature regulation center (such as hypothalamic damage, peripheral vasodilation and hypotension, etc.), leading to failure of heat dissipation mechanisms (such as sweating, vasodilation) and insufficient circulating blood volume; second, imbalance between heat production and heat dissipation (such as excessive heat production, heat dissipation disorders), leading to rapid increase in heat production and heat accumulation; third, systemic inflammatory response and oxidative stress (such as cytokine storm, oxidative stress injury, etc.), leading to systemic inflammatory response syndrome (SIRS) and exacerbating multiple organ injury; fourth, endothelial cell damage and coagulation dysfunction (such as endothelial damage, disseminated intravascular coagulation (DIC)), leading to tissue edema and microthrombosis, resulting in bleeding or embolism; fifth, organ dysfunction (such as central nervous system damage, rhabdomyolysis, circulatory collapse, liver and kidney function damage), leading to acute kidney injury (AKI), arrhythmia, heart failure, etc.

[0003] Heatstroke is divided into mild heatstroke (body temperature of 38-40℃, with symptoms such as dizziness, nausea, and sweating), moderate heatstroke (body temperature close to 40℃, with symptoms such as confusion, blood pressure drop, etc.), and severe heatstroke (core body temperature > 40℃, with symptoms such as convulsions, coma, multiple organ failure, etc., with a mortality rate of 10%-50%).

[0004] Heatstroke is divided into classic heatstroke (non-exertional heatstroke), exertional heatstroke (EHS), and other types (such as pediatric heatstroke, drug or disease-related heatstroke, and malignant hyperthermia, etc.) according to etiology and pathogenesis.

[0005] Older people (especially those with chronic diseases such as heart failure, diabetes, cognitive impairment), infants, obese people, long-term bedridden patients, etc., are prone to non-exertional heatstroke due to failure of body temperature regulation in high temperature, high humidity, closed, poorly ventilated environments (such as rooms without air conditioning). Non-exertional heatstroke has the characteristics of insidious onset and multiple complications, often accompanied by dehydration, electrolyte imbalance, heart and kidney dysfunction, etc.

[0006] The main causes of exertional heat stroke include excessive heat production caused by high-intensity physical activity (such as exercise, military training, heavy physical labor), which exceeds the body's heat dissipation capacity. The high-risk groups of exertional heat stroke include athletes, soldiers, firefighters, construction workers, and other healthy individuals who are not adapted to high-temperature environments (such as new recruits). Exertional heat stroke has the characteristics of rapid onset (rapid deterioration within a few hours), early onset of multiple organ damage (such as the occurrence of rhabdomyolysis, acute kidney injury, liver failure, disseminated intravascular coagulation (DIC), etc.), and rapid body temperature rise (often exceeding 40.5℃, even reaching 42℃). Patients develop central nervous system dysfunction, leaving brain dysfunction and muscle damage, and the mortality rate is 20%-70%. Moreover, if the patient's body temperature is ≥40.5℃ for more than 1 hour, the mortality rate will significantly increase, and the clinical emphasis is on the “golden 1 hour” intervention.

[0007] The key to the treatment of exertional heat stroke is to rapidly reduce the patient's core body temperature and inhibit the inflammatory factor storm, combined with fluid resuscitation, electrolyte correction, organ and function protection, anticoagulation and anti-inflammatory treatment, and management of complications (such as acute kidney injury, rhabdomyolysis, and infection prevention). It requires a therapeutic drug with rapid release and onset, multi-organ (such as brain, liver, and kidney) targeting, etc., to reduce the damage and mortality rate caused by heat stroke to patients.

[0008] Previous studies have shown that the intestinal epithelium is considered the most critical barrier against harmful antigens and pathogens. Oxidative stress in the body can lead to intestinal barrier dysfunction. Under exercise and heat stress, the body's intestinal epithelium and intestinal permeability increase, and endotoxins are translocated to the circulatory system, leading to persistent heat stress, etc., allowing bacterial lipopolysaccharide to enter the portal circulation, which exceeds the clearance capacity of the liver and immune system, and then lipopolysaccharide enters the systemic circulation to cause endotoxemia, leading to acute inflammatory response, diffuse intravascular coagulation (DIC), MODs, and other heat stroke symptoms. Excessive exercise suppresses the immune system and reduces the body's clearance of lipopolysaccharide, exacerbating the damage to multiple organ systems caused by heat stroke. Preventing damage to the intestinal mucosal barrier and maintaining the integrity of the intestinal barrier can effectively prevent the translocation of intestinal bacteria and toxins to capillaries, prevent heat stroke, and reduce mortality.

[0009] The difficulty in the treatment of heat stroke is the lethality of delayed cooling and the resulting multiple organ failure, but existing drug treatment is mainly symptomatic. Therefore, it is necessary to develop new targeted drugs for the pathogenesis and pathological characteristics of heat stroke (such as high fever, oxidative stress, inflammatory response, and multiple organ damage).

[0010] Curcumin has antibacterial, antioxidant, anti-inflammatory, anticancer, antiviral, antidiabetic, anti-allergic asthma, etc. effects, and is used to reduce the damage of heat stroke to multiple organs (such as heart, lung, liver, kidney, intestine, brain, etc.) and its secondary damage, reduce the expression of inflammatory factors, etc. However, curcumin is a poorly soluble drug, and has pH dependence (rapid degradation at pH>7), and is degraded due to sensitivity to light, heat (high temperature (>60℃)) which leads to intramolecular dehydration or ring-opening reaction, and its stability and bioavailability need to be improved.

[0011] (Curcumin)

[0012] Document 1 (CN116172990A) discloses a curcumin nanocrystal suspension, which has the following defects: first, the concentration of curcumin in the nanocrystal suspension is low (such as 2.7%); second, in order to improve the solubility of curcumin, the average particle size of curcumin nanocrystals in the nanocrystal suspension is ≤100 nm (such as 71 nm), which leads to a longer preparation time of the suspension; third, a 6% (m / v) sucrose solution is needed in the preparation to improve the stability of the nanocrystal suspension; fourth, the nanocrystal suspension is not convenient to carry, etc.

[0013] Document 2 (Peng YF et al., Preparation and in vitro and in vivo evaluation of curcumin oral nanocrystal capsules, Journal of China Pharmaceutical University, 2021, 52(2): 211-218) discloses drug-loaded pellets and capsules prepared by solidifying curcumin nanocrystal suspension, which preliminarily solves the problem of inconvenience of carrying curcumin nanocrystal suspension, but still has the following defects: first, the concentration of curcumin in the nanocrystal suspension is low (such as 10%), which leads to a low amount of curcumin loaded by the drug-loaded pellets; second, the average particle size of curcumin nanocrystals is about 200 nm, and the preparation time is long; third, the nanocrystal suspension needs to be mixed with an equal amount of sucrose before being used to solidify and prepare drug-loaded pellets in order to improve its stability; fourth, the flowability of the drug-loaded pellets needs to be improved, etc.

[0014] Piperine ((1E,2E)-1-(1,3-benzodioxol-5-yl)-3-(1-propen-3-yl)-1,3-dihydroisoquinoline-2-one, C 17 H 19Piperine (C17H19NO3) is the main bioactive component extracted from black pepper (Piper nigrum), which has the effects of promoting the absorption of drugs / nutrients, anti-platelet aggregation, anti-inflammatory activity, antioxidant activity, neuroprotective activity, antibacterial, regulating intestinal flora, anti-fatigue, etc. Piperine is slightly soluble in water, and is prone to photo-oxidation under ultraviolet (UV) irradiation to generate degradation products such as dehydro-piperine. It starts to decompose at >80°C, is prone to hydrolysis under neutral / alkaline conditions (pH >7), and is prone to oxidative degradation when in contact with oxygen. It is prone to moisture absorption and caking in a high-humidity (>60%) environment. Therefore, it is necessary to improve its solubility, stability and bioavailability, etc.

[0015] In order to effectively prevent and treat exertional heat illness and the multiple organ system damage caused thereby, it is necessary to develop a rapid-acting targeted drug to meet the clinical needs. SUMMARY

[0016] The present application aims to provide a curcumin nanocrystal composition made from a curcumin nanocrystal suspension and a piperine suspension, wherein the mass ratio of curcumin:piperine is 5-15:1, preferably 8-12:1, and more preferably 10:1.

[0017] In a preferred technical solution of the present application, the curcumin nanocrystal suspension contains 20-35% curcumin nanocrystals, 1-10% steric protective agent and 1-10% charge protective agent, wherein the average particle size of the curcumin nanocrystals is 200-320 nm.

[0018] In a preferred technical solution of the present application, the average particle size of the curcumin nanocrystals is 220-300 nm, and preferably 250-280 nm.

[0019] In a preferred technical solution of the present application, the steric protective agent is selected from any one or a combination of poloxamer, Tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124 and poloxamer 188.

[0020] In a preferred technical solution of the present application, the charge protective agent is selected from any one or a combination of sodium dodecyl sulfate (SDS), sodium deoxycholate and sodium docusate.

[0021] In a preferred technical solution of the present application, the curcumin nanocrystal suspension contains 20-35% curcumin nanocrystals, 1-10% poloxamer 407 and 1-10% sodium dodecyl sulfate in terms of mass volume percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, and preferably 250-280 nm.

[0022] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 30% of curcumin nanocrystals, 5-9% of poloxamer 407 and 1.5-5% of sodium dodecyl sulfate in terms of mass volume percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0023] In the preferred technical scheme of the present application, the preparation of the curcumin nanocrystal suspension comprises the following steps: dissolving a required amount of a steric protective agent and a charge protective agent in water, then adding a required amount of curcumin, controlling the sample feeding speed to be 150-300 g / min, and feeding it into a grinder for grinding ((2000-3500 rpm)*(15-60 min)) to obtain the curcumin nanocrystal suspension.

[0024] In the preferred technical scheme of the present application, the sample feeding speed is 200-250 g / min.

[0025] In the preferred technical scheme of the present application, the grinding condition is (2500-3000 rpm)*(30-50 min).

[0026] Another object of the present application is to provide a preparation method of a curcumin nanocrystal composition,

[0027] The curcumin nanocrystal composition is prepared from the curcumin nanocrystal suspension and the piperine suspension, wherein the mass ratio of curcumin to piperine is 5-15:1, and the preparation method of the curcumin nanocrystal composition is mixing the curcumin nanocrystal suspension and the piperine suspension according to the mass ratio of curcumin to piperine being 5-15:1.

[0028] In the preferred technical scheme of the present application, the mass ratio of curcumin to piperine is 8-12:1, preferably 10:1.

[0029] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 20-35% of curcumin nanocrystals, 1-10% of a steric protective agent and 1-10% of a charge protective agent, wherein the average particle size of the curcumin nanocrystals is 200-320 nm.

[0030] In the preferred technical scheme of the present application, the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0031] In the preferred technical scheme of the present application, the steric protective agent is selected from any one or a combination of poloxamer, Tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124 and poloxamer 188.

[0032] In the preferred technical solution of the present application, the charge protective agent is selected from any one or combination of sodium dodecyl sulfate (SDS), sodium deoxycholate and sodium docusate.

[0033] In the preferred technical solution of the present application, the curcumin nanocrystal suspension contains, in terms of mass volume percentage, 20-35% curcumin nanocrystals, 1-10% poloxamer 407 and 1-10% sodium dodecyl sulfate, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0034] In the preferred technical solution of the present application, the curcumin nanocrystal suspension contains, in terms of mass volume percentage, 30% curcumin nanocrystals, 5-9% poloxamer 407 and 1.5-5% sodium dodecyl sulfate, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0035] In the preferred technical solution of the present application, the preparation of the curcumin nanocrystal suspension comprises the following steps: dissolving a required amount of the steric protective agent and the charge protective agent in water, adding a required amount of curcumin, controlling the sample feeding speed to be 150-300 g / min, and feeding it into a grinder for grinding ((2000-3500 rpm)*(15-60 min)) to obtain the curcumin nanocrystal suspension.

[0036] In the preferred technical solution of the present application, the sample feeding speed is 200-250 g / min.

[0037] In the preferred technical solution of the present application, the grinding condition is (2500-3000 rpm)*(30-50 min).

[0038] The present application aims to provide a curcumin nanocrystal composition, which is prepared by drying and solidifying the curcumin nanocrystal suspension onto a blank core, wherein the curcumin nanocrystal suspension contains, in terms of mass volume percentage, 20-35% curcumin nanocrystals, 1-10% steric protective agent and 1-10% charge protective agent, and the average particle size of the curcumin nanocrystals is 200-320 nm.

[0039] In the preferred technical solution of the present application, the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0040] In the preferred technical solution of the present application, the steric protective agent is selected from any one or combination of poloxamer, Tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124 and poloxamer 188.

[0041] In the preferred technical scheme of the present application, the charge protective agent is selected from any one or combination of sodium dodecyl sulfate (SDS), sodium deoxycholate and sodium docusate.

[0042] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 20-35% curcumin nanocrystals, 1-10% poloxamer 407 and 1-10% sodium dodecyl sulfate by mass percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0043] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 30% curcumin nanocrystals, 5-9% poloxamer 407 and 1.5-5% sodium dodecyl sulfate by mass percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0044] In the preferred technical scheme of the present application, the preparation of the curcumin nanocrystal suspension comprises the following steps: dissolving a required amount of the steric protective agent and the charge protective agent in water, adding a required amount of curcumin, controlling the sample feeding speed to be 150-300 g / min, and feeding it into a grinder for grinding ((2000-3500 rpm)*(15-60 min)) to obtain the curcumin nanocrystal suspension.

[0045] In the preferred technical scheme of the present application, the sample feeding speed is 200-250 g / min.

[0046] In the preferred technical scheme of the present application, the grinding condition is (2500-3000 rpm)*(15-30 min).

[0047] In the preferred technical scheme of the present application, the preparation of the curcumin nanocrystal composition comprises the following steps:

[0048] (1) preheat the fluidized bed to a set temperature of 40-60℃, and feed in the blank pellet cores;

[0049] (2) under the conditions of an inlet air temperature of 45-55℃, an atomization pressure of 1.8-2.2 bar, a liquid spraying speed of 60-90 g / h and an inlet air volume of 30-55 m³ / h, spray the curcumin nanocrystal suspension through an atomizer onto the fluidized blank pellet cores;

[0050] (3) after the liquid spraying is completed, perform air drying (40-70℃)*(10-40 min) to dry and solidify the suspension on the blank pellet cores to obtain the drug-loaded pellets.

[0051] In the preferred technical scheme of the present application, the water content of the blank pellet cores in step (1) is <5%.

[0052] In the preferred technical scheme of the present application, the drying condition in step (3) is (50-60℃) *(20-30min).

[0053] In the preferred technical scheme of the present application, the particle size of the blank pellet core is 0.3-0.5mm.

[0054] In the preferred technical scheme of the present application, the blank pellet core is selected from any one of cellulose pellet core, lactose pellet core.

[0055] In the preferred technical scheme of the present application, the cellulose pellet core is selected from any one of microcrystalline cellulose blank pellet core, Avicel PH-101 / PH-102 blank pellet core, Avicel PH-301 / PH-302 blank pellet core, hydroxypropyl methyl cellulose (HPMC) blank pellet core, Methocel E5 / E15, sodium carboxymethyl cellulose (CMC-Na) blank pellet core, low-substituted CMC-Na blank pellet core, high-substituted CMC-Na blank pellet core, methyl cellulose (MC) blank pellet core, MC-L (low viscosity) methyl cellulose blank pellet core, MC-H (high viscosity) methyl cellulose blank pellet core, microcrystalline cellulose + lactose (such as Cellactose 80) blank pellet core, microcrystalline cellulose + spray-dried lactose (such as Prosolv SMCC) blank pellet core or a combination thereof.

[0056] In the preferred technical scheme of the present application, the curcumin nanocrystal composition optionally contains piperine.

[0057] In the preferred technical scheme of the present application, the mass ratio of curcumin to piperine in the curcumin nanocrystal composition is 5-15:1-2, preferably 8-12:1-2, and more preferably 10:1.

[0058] In the preferred technical scheme of the present application, the combination of curcumin nanocrystal and piperine is selected from any one of combination, simultaneous administration, and sequential administration.

[0059] In the preferred technical scheme of the present application, the composition is an oral preparation.

[0060] In the preferred technical scheme of the present application, the oral preparation is selected from any one of capsule, tablet, and granule.

[0061] In the preferred technical scheme of the present application, the prepared curcumin nanocrystal composition or curcumin nanocrystal drug-loaded pellet is filled into a capsule to prepare a curcumin nanocrystal capsule.

[0062] Another object of the present application is to provide a preparation method of a curcumin nanocrystal composition, which is prepared by drying and solidifying a curcumin nanocrystal suspension onto a blank core, wherein the curcumin nanocrystal suspension contains 20-35% of curcumin nanocrystals, 1-10% of a steric protective agent and 1-10% of a charge protective agent by mass percentage, and wherein the average particle size of the curcumin nanocrystals is 200-320 nm, and the preparation of the curcumin nanocrystal composition comprises the following steps:

[0063] (1) preheat the fluidized bed to a set temperature of 40-60°C, and then put in the blank core;

[0064] (2) under the conditions of an inlet air temperature of 45-55°C, an atomization pressure of 1.8-2.2 bar, a liquid spraying speed of 60-90 g / h, and an inlet air volume of 30-55 m³ / h, spray the curcumin nanocrystal suspension through an atomizer onto the fluidized blank core;

[0065] (3) after the spraying is completed, perform air drying (40-70°C) for (10-40) min to dry and solidify the suspension on the blank core, thereby preparing the drug-loaded pellets.

[0066] In a preferred technical solution of the present application, the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0067] In a preferred technical solution of the present application, the steric protective agent is selected from any one or a combination of poloxamer, Tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124 and poloxamer 188.

[0068] In a preferred technical solution of the present application, the charge protective agent is selected from any one or a combination of sodium dodecyl sulfate (SDS), sodium deoxycholate and sodium docusate.

[0069] In a preferred technical solution of the present application, the curcumin nanocrystal suspension contains 20-35% of curcumin nanocrystals, 1-10% of poloxamer 407 and 1-10% of sodium dodecyl sulfate by mass percentage, and the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0070] In a preferred technical solution of the present application, the curcumin nanocrystal suspension contains 30% of curcumin nanocrystals, 5-9% of poloxamer 407 and 1.5-5% of sodium dodecyl sulfate by mass percentage, and the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0071] In the preferred technical scheme of the present application, the preparation of the curcumin nanocrystal suspension comprises the following steps: dissolving a required amount of steric protection agent and charge protection agent in water, then adding a required amount of curcumin, controlling the sample injection speed to be 150-300 g / min, and grinding it in a grinder ((2000-3500 rpm)*(15-60 min)) to obtain the curcumin nanocrystal suspension.

[0072] In the preferred technical scheme of the present application, the sample injection speed is 200-250 g / min.

[0073] In the preferred technical scheme of the present application, the grinding condition is (2500-3000 rpm)*(15-30 min).

[0074] In the preferred technical scheme of the present application, the water content of the blank pellet core in step (1) is <5%.

[0075] In the preferred technical scheme of the present application, the drying condition in step (3) is (50-60 ℃)*(20-30 min).

[0076] In the preferred technical scheme of the present application, the particle size of the blank pellet core is 0.3-0.5 mm.

[0077] In the preferred technical scheme of the present application, the blank pellet core is selected from any one of cellulose pellet cores, lactose pellet cores.

[0078] In the preferred technical scheme of the present application, the cellulose pellet core is selected from any one of microcrystalline cellulose blank pellet core, Avicel PH-101 / PH-102 blank pellet core, Avicel PH-301 / PH-302 blank pellet core, hydroxypropyl methyl cellulose (HPMC) blank pellet core, Methocel E5 / E15, sodium carboxymethyl cellulose (CMC-Na) blank pellet core, low-substituted CMC-Na blank pellet core, high-substituted CMC-Na blank pellet core, methyl cellulose (MC) blank pellet core, MC-L (low viscosity) methyl cellulose blank pellet core, MC-H (high viscosity) methyl cellulose blank pellet core, microcrystalline cellulose+lactose (such as Cellactose 80) blank pellet core, microcrystalline cellulose+spray-dried lactose (such as Prosolv SMCC) blank pellet core, or a combination thereof.

[0079] In the preferred technical scheme of the present application, the curcumin nanocrystal composition optionally contains piperine.

[0080] In the preferred technical scheme of the present application, the mass ratio of curcumin to piperine in the curcumin nanocrystal composition is 5-15:1-2, preferably 8-12:1-2, and more preferably 10:1.

[0081] In the preferred technical scheme of the present application, the combination of curcumin nanocrystals and piperine is selected from any one of the following: a composition, simultaneous administration, and sequential administration.

[0082] In the preferred technical scheme of the present application, the composition is an oral preparation.

[0083] In the preferred technical scheme of the present application, the oral preparation is selected from any one of the following: a capsule, a tablet, and a granule.

[0084] In the preferred technical scheme of the present application, the prepared curcumin nanocrystal composition or curcumin nanocrystal drug-loaded pellet is filled into a capsule to obtain a curcumin nanocrystal capsule.

[0085] The present application aims to provide a curcumin nanocrystal suspension, which contains 20-35% curcumin nanocrystals, 1-10% steric protective agent, and 1-10% charge protective agent, wherein the average particle size of the curcumin nanocrystals is 200-320 nm.

[0086] In the preferred technical scheme of the present application, the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0087] In the preferred technical scheme of the present application, the steric protective agent is selected from any one of the following: poloxamer, Tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124, and poloxamer 188, or a combination thereof.

[0088] In the preferred technical scheme of the present application, the charge protective agent is selected from any one of the following: sodium dodecyl sulfate (SDS), sodium deoxycholate, and sodium docusate, or a combination thereof.

[0089] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 20-35% curcumin nanocrystals, 1-10% poloxamer 407, and 1-10% sodium dodecyl sulfate, in terms of mass percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0090] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 30% curcumin nanocrystals, 5-9% poloxamer 407, and 1.5-5% sodium dodecyl sulfate, in terms of mass percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0091] In the preferred technical scheme of the present application, the preparation of the curcumin nanocrystal suspension comprises the following steps: dissolving a required amount of the steric protective agent and the charge protective agent in water, then adding a required amount of curcumin, controlling the sample feeding speed to be 150-300 g / min, and feeding it into a grinder for grinding ((2000-3500 rpm)*(15-60 min)) to obtain the curcumin nanocrystal suspension.

[0092] In the preferred technical scheme of the present application, the sample feeding speed is 200-250 g / min.

[0093] In the preferred technical scheme of the present application, the grinding condition is (2500-3000 rpm)*(15-30 min).

[0094] Another object of the present application is to provide a preparation method of a curcumin nanocrystal suspension, the curcumin nanocrystal suspension containing 20-35% of curcumin nanocrystals, 1-10% of a steric protective agent and 1-10% of a charge protective agent, wherein the average particle size of the curcumin nanocrystals is 200-320 nm, and the preparation of the curcumin nanocrystal suspension comprises the following steps: dissolving a required amount of the steric protective agent and the charge protective agent in water, then adding a required amount of curcumin, controlling the sample feeding speed to be 150-300 g / min, and feeding it into a grinder for grinding ((2000-3500 rpm)*(15-60 min)) to obtain the curcumin nanocrystal suspension.

[0095] In the preferred technical scheme of the present application, the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0096] In the preferred technical scheme of the present application, the steric protective agent is selected from any one or combination of poloxamer, tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124 and poloxamer 188.

[0097] In the preferred technical scheme of the present application, the charge protective agent is selected from any one or combination of sodium dodecyl sulfate (SDS), sodium deoxycholate and sodium docusate.

[0098] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 20-35% of curcumin nanocrystals, 1-10% of poloxamer 407 and 1-10% of sodium dodecyl sulfate in terms of mass volume percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0099] In the preferred technical scheme of the present application, the curcumin nanocrystal suspension contains 30% curcumin nanocrystals, 5-9% poloxamer 407 and 1.5-5% sodium dodecyl sulfate in terms of mass volume percentage, wherein the average particle size of the curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

[0100] In the preferred technical scheme of the present application, the sample injection speed is 200-250 g / min.

[0101] In the preferred technical scheme of the present application, the grinding condition is (2500-3000 rpm)*(15-30 min).

[0102] Another object of the present application is to provide the use of the curcumin nanocrystal composition or the curcumin nanocrystal suspension of the present application for preparing a medicament for preventing and treating exertional heat stroke and multiple organ injury caused thereby or complications thereof.

[0103] Unless otherwise specified, when the present application relates to the percentage between liquids, the percentage is volume / volume percentage; when the present application relates to the percentage between liquids and solids, the percentage is volume / weight percentage; when the present application relates to the percentage between solids and liquids, the percentage is weight / volume percentage; and the rest is weight / weight percentage.

[0104] Unless otherwise specified, the particle size of the nanocrystals is detected by a nanoparticle size analyzer (Nano-ZS90, Malvern, UK).

[0105] The content of curcumin is detected by high performance liquid chromatography: the chromatographic column is Aglient TC-C18 (250 mm*4.6 mm, 5 μm); the mobile phase is acetonitrile:4% glacial acetic acid solution (40:60); the flow rate is 1.0 mL / min; the column temperature is 30℃; the wavelength is 264 nm; and the injection amount is 10 μL. 20 mg of curcumin nanocrystal pellets are taken, placed in a 100 mL volumetric flask, diluted with water to the calibration mark, and shaken to obtain the curcumin nanocrystal suspension.

[0106] The blood drug concentration is determined by high performance liquid chromatography-mass spectrometry, and the specific reference is reference 2 (Peng Yifan et al., Preparation and in vitro and in vivo evaluation of curcumin oral nanocrystal capsules, Journal of China Pharmaceutical University, 2021, 52(2):211-218).

[0107] Compared with the prior art, the present application has the following beneficial technical effects:

[0108] 1、The present application scientifically screens the components and the ratio of curcumin nanocrystals, and prepares drug-loaded pellets, thereby improving the drug loading concentration of curcumin in the nanocrystal suspension, the average particle size of the curcumin nanocrystals is 200-320 nm, the preparation time of the nanocrystal suspension is shortened, sucrose is omitted as a protective agent, and the stability of the nanocrystals is improved. The curcumin nanocrystal composition prepared by the present application significantly improves heat tolerance, prolongs survival time, reduces the level of inflammatory factors in the blood, protects intestinal epithelial cells and intestinal barrier function, reduces heat stroke injury, can effectively prevent and treat heat stroke and its complications, prevent and treat the damage of the body and organs caused by heat stroke, and has the advantages of high drug loading, good stability, high bioavailability, safety and effectiveness, easy to carry, etc.

[0109] 2、The curcumin nanocrystals of the present application are used in combination with piperine to synergistically improve heat tolerance and have better effect of improving and preventing heat stroke.

[0110] 3、The preparation process of the present application has the advantages of simple operation, significantly shortened production cycle, high yield, green environmental protection, significant cost benefit, and is suitable for industrial mass production. BRIEF DESCRIPTION OF DRAWINGS

[0111] Figure 1 The shape of the curcumin nanocrystal composition of the present application is characterized;

[0112] Figure 2 The effect of the curcumin nanocrystals of the present application on preventing and treating labor heat stroke is studied;

[0113] Figure 3 The influence of the curcumin nanocrystals of the present application on intestinal permeability of labor heat stroke rats;

[0114] Figure 4 The effect of the curcumin nanocrystals of the present application on improving apoptosis of intestinal epithelial cells of labor heat stroke rats is studied;

[0115] Figure 5 The effect of the curcumin nanocrystals of the present application on reducing the whole blood IL-6 level of labor heat stroke rats is studied;

[0116] Figure 6 The protective effect of the curcumin nanocrystals of the present application on damaged intestines and lungs of labor heat stroke rats is studied;

[0117] Figure 7 The stability of the curcumin nanocrystal composition of the present application is investigated;

[0118] Figure 8 The pharmacokinetics of the curcumin nanocrystal composition of the present application is studied. DETAILED DESCRIPTION

[0119] The present application is illustrated below with reference to examples, but the present application is not limited to the examples.

[0120] Ball mill (WAB, Switzerland, model ECM-AP 05).

[0121] Fluidized bed (Chongqing Yingge Granulation and Coating Technology Co., Ltd., model WRTFL-60)

[0122] The water content of the sodium carboxymethylcellulose (CMC-Na) blank pellet core (particle size 0.3-0.5 mm) is less than 5%.

[0123] Preparation of 30% (m / v) curcumin suspension in the specific embodiment: 45 g of poloxamer 407 and 7.5 g of sodium dodecyl sulfate (SDS) were dissolved in 500 mL of water, and then 150 g of curcumin was added, stirred, and mixed uniformly to obtain the suspension.

[0124] Preparation of 30% (m / v) piperine suspension in the specific embodiment: 30 g of piperine was dispersed in 100 mL of water to obtain the suspension.

[0125] Example 1 Preparation of curcumin nanocrystal suspension

[0126] 45 g of poloxamer 407 and 7.5 g of sodium dodecyl sulfate were dissolved in 500 mL of water, and then 150 g of curcumin was added, and the feeding speed was controlled at 250 g / min, and then it was put into the grinder for grinding (3000 rpm*5 min) to obtain a curcumin nanocrystal suspension with an average particle size of 380 nm.

[0127] Example 2 Preparation of curcumin nanocrystal suspension

[0128] 45 g of poloxamer 407 and 7.5 g of sodium dodecyl sulfate were dissolved in 500 mL of water, and then 150 g of curcumin was added, and the feeding speed was controlled at 250 g / min, and then it was put into the grinder for grinding (3000 rpm*5 min) to obtain a curcumin nanocrystal suspension with an average particle size of 380 nm.

[0129] Example 3 Preparation of curcumin nanocrystal suspension

[0130] 45 g of poloxamer P407 and 7.5 g of sodium dodecyl sulfate were dissolved in 500 mL of water, and then 150 g of curcumin was added, and the feeding speed was controlled at 250 g / min, and then it was put into the grinder for grinding (3000 rpm*5 min) to obtain a curcumin nanocrystal suspension with an average particle size of 380 nm.

[0131] Example 4 Preparation of curcumin nanocrystal composition

[0132] Preparation of the curcumin nanocrystal composition comprises the following steps:

[0133] (1) Dissolve 45 g of poloxamer P188 and 7.5 g of sodium dodecyl sulfate in 500 mL of water, then add 150 g of curcumin, control the feeding speed to be 250 g / min, and put it into a grinder (3000 rpm*15 min) to grind, to obtain a curcumin nanocrystal suspension with an average particle size of 280 nm.

[0134] (2) Preheat the fluidized bed to 45℃, the atomization pressure is 1.8-2.0 bar, the air inlet volume is 55 m³ / h, the air inlet temperature is 45℃, and the peristaltic pump speed is 1.6 rpm; first put in 50 g of sodium carboxymethyl cellulose blank pellet cores, then spray the curcumin nanocrystal suspension prepared in step (1) at a liquid spraying speed of 85 g / h, dry at 60℃ for 10 min, to obtain curcumin nanocrystal drug-loaded pellets, which are loaded into a 0 size capsule shell (300 mg / pellet capsule). Figure 1 .

[0135] Example 5 Preparation of the curcumin nanocrystal composition of the present application

[0136] Preparation of the curcumin nanocrystal composition comprises the following steps:

[0137] (1) Dissolve 45 g of poloxamer P188 and 7.5 g of sodium dodecyl sulfate in 500 mL of water, then add 150 g of curcumin, control the feeding speed to be 250 g / min, and put it into a grinder (3000 rpm*15 min) to grind, to obtain a curcumin nanocrystal suspension with an average particle size of 280 nm.

[0138] (2) Preheat the fluidized bed to 45℃, the atomization pressure is 1.8-2.0 bar, the air inlet volume is 55 m³ / h, the air inlet temperature is 45℃, and the peristaltic pump speed is 1.6 rpm; first put in 50 g of sodium carboxymethyl cellulose blank pellet cores, then spray the curcumin nanocrystal suspension prepared in step (1) at a liquid spraying speed of 85 g / h, dry at 60℃ for 10 min, to obtain curcumin nanocrystal drug-loaded pellets, which are loaded into a 0 size capsule shell (300 mg / pellet capsule).

[0139] Example 6 Preparation of the curcumin nanocrystal composition of the present application

[0140] Preparation of the curcumin nanocrystal composition comprises the following steps:

[0141] Dissolve 45g poloxamer P188 and 7.5g sodium dodecyl sulfate in 500mL of water, then add 150g curcumin. Control the injection speed at 250g / min and grind the mixture in a grinder (3000rpm*15min) to obtain a curcumin nanocrystal suspension with an average particle size of 280nm.

[0142] (2) Preheat the fluidized bed to 45°C, atomize at 1.8-2.0 bar, inlet air volume at 55 m³ / h, inlet air temperature at 45°C, and peristaltic pump at 1.6 rpm. First, add 50 g of lactose blank pellet core, then spray the curcumin nanocrystal suspension obtained in step (1) at a spraying speed of 85 g / h. Dry at 60°C for 10 min to obtain curcumin nanocrystal drug-loaded microspheres, and fill them into No. 0 capsule shells (300 mg / capsule).

[0143] Example 7 Preparation of the Curcumin Nanocrystal Composition of the Present Invention

[0144] Dissolve 45g of poloxamer 407 and 7.5g of sodium dodecyl sulfate (SDS) in 500mL of water, then add 150g of curcumin and grind it in a grinder (3000rpm*5min) to obtain a curcumin nanocrystal suspension with an average particle size of 380nm. Then add 30% (m / v) of piperine suspension at a mass ratio of curcumin nanocrystals to piperine of 10:1 and mix well to obtain the final product.

[0145] Example 8 Preparation of Curcumin Nanocrystal Piperine Mixture of the Invention

[0146] Dissolve 45g of poloxamer 407 and 7.5g of sodium dodecyl sulfate (SDS) in 500mL of water, then add 150g of curcumin and grind in a grinder (3000rpm*15min) to obtain a curcumin nanocrystal suspension with an average particle size of 280nm. Then add 30% (m / v) of piperine suspension at a mass ratio of curcumin nanocrystals to piperine of 10:1 and mix evenly to obtain the final product.

[0147] Example 9 Preparation of Curcumin Nanocrystal Piperine Mixture of the Invention

[0148] Dissolve 45g of poloxamer 407 and 7.5g of sodium dodecyl sulfate (SDS) in 500mL of water, then add 150g of curcumin and grind in a grinder (3000rpm*60min) to obtain a curcumin nanocrystal suspension with an average particle size of 75nm. Then add 30% (m / v) of piperine suspension at a mass ratio of curcumin nanocrystals to piperine of 10:1 and mix evenly to obtain the final product.

[0149] Test Example 1 Establishment of an experimental model of labor-induced heatstroke

[0150] Thirty male SD rats weighing 200-220g (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were randomly divided into three groups of 10 each. The animals were subjected to heat shock in a closed environment at 42℃ and 50% humidity. The rectal temperature of the animals was measured and recorded every 30 minutes.

[0151] After 150 minutes of heat shock, the body temperature of the experimental animals rose to approximately 40.5°C. The temperature initially rose rapidly, then decreased after the body regulated itself, before resuming a rapid rise. This indicates that the labor-induced heatstroke experimental model was successfully established. No statistically significant differences were found among the experimental animals.

[0152] Test Example 2 Research on the effect of curcumin nanocrystals in preventing and treating labor-related heatstroke.

[0153] One hundred male SD rats weighing 200-220g (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were randomly divided into 10 groups of 10 rats each. During the experiment, the animals were fed and hydrated normally. The animals were administered the drug or an equal volume of water by gavage at a dose of 200mg / kg daily for seven consecutive days.

[0154] Control group: Gavaged with an equal volume of water;

[0155] Control group 1: Curcumin suspension administered by gavage;

[0156] Control group 2: Piperine suspension administered by gavage;

[0157] Experimental group 1: Gavage administration of curcumin nanocrystal suspension from Example 1;

[0158] Experimental group 2: Gavage administration of curcumin nanocrystal suspension from Example 2;

[0159] Experimental group 3: Gavage administration of curcumin nanocrystal suspension from Example 3;

[0160] Experimental group 4: Curcumin nanocrystal composition of Example 4 administered by gavage;

[0161] Experimental group 5: Curcumin nanocrystal composition of Example 7 administered by gavage;

[0162] Experimental group 6: Curcumin nanocrystal composition of Example 8 administered by gavage;

[0163] Experimental group 7: Curcumin nanocrystal composition of Example 9 administered by gavage.

[0164] After the seventh day of gavage administration, following the modeling method in Example 1, the experimental animals were subjected to heat shock in a closed environment at 42°C and 50% humidity. Every 30 minutes, the rectal temperature and the time taken for the rectal temperature to reach 40.5°C were measured and recorded. Results are shown below. Figure 2 .

[0165] Curcumin nanocrystals with an average particle size of 280 nm and curcumin nanocrystals with an average particle size of 75 nm showed essentially the same effect on the prevention and treatment of labor-related heatstroke, with no significant difference between the two.

[0166] Test Example 3 This invention studies the protective effect of curcumin nanocrystals against intestinal permeability and multi-organ damage in patients with exertional heatstroke.

[0167] Forty male SD rats weighing 200-220g (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were randomly divided into four groups of 10 rats each. During the experiment, the animals were fed and hydrated normally. The animals were administered the drug or an equal volume of water by gavage at a dose of 200mg / kg daily for seven consecutive days.

[0168] Control group: Gavaged with an equal volume of water;

[0169] Control group: Curcumin suspension administered by gavage;

[0170] Experimental group 1: Gavage administration of curcumin nanocrystal suspension from Example 2;

[0171] Experimental group 2: Gavage administration of curcumin nanocrystal suspension from Example 3.

[0172] After the seventh day of gavage administration, the experimental animals were administered 4-kDa FITC-glucan (40 mg / kg; Beyotime, China) by gavage for 30 min. Following the modeling method in Example 1, the animals were subjected to heat shock in a closed environment at 42°C and 50% humidity for 2 h. Blood was then collected from the abdominal aorta, and the FITC fluorescence of the plasma was measured using a microplate reader with 488nm excitation / 535nm emission. The plasma FITC-glucan value was calculated based on the FITC-glucan standard curve. Results are shown below. Figure 3 .

[0173] Two hours after heat shock, the ileum of the experimental animals was removed for TUNEL staining to observe intestinal epithelial cell apoptosis. Results are shown below. Figure 4 .

[0174] Two hours after heat shock, whole blood was collected from the experimental animals, and the level of interleukin-6 (IL-6) was measured. The results are shown below. Figure 5 .

[0175] After 2 hours of heat shock, intestinal tissue was taken from the experimental animals (see...).Figure 6-1 ) and lung tissue (see Figure 6-2 HE staining was performed, and the results are shown in Figure 6, where (a) is the blank group; (b) is the control group; (c) is the experimental group 1; and (d) is the experimental group 2.

[0176] Curcumin nanocrystals with an average particle size of 280 nm and those with an average particle size of 75 nm can both protect intestinal epithelial cells, reduce intestinal barrier permeability and systemic inflammation levels, control immune dysregulation, and significantly improve multi-organ damage caused by exertional heatstroke. Moreover, the two have essentially the same effect in preventing and treating heatstroke and its multi-organ damage, and no significant difference was found between them.

[0177] Test Example 4 Stability study of the curcumin nanocrystal composition of the present invention

[0178] Using the curcumin nanocrystal composition of Example 4 as an example, its stability was investigated by placing it under high temperature (60℃±2℃) and light (5000Lx±500Lx) conditions for 30 days. The results are shown in […]. Figure 7 .

[0179] Test Example 5 Pharmacokinetic Study of the Curcumin Nanocrystal Composition of the Invention

[0180] Thirty male SD rats (Beijing Vital River Laboratory Animal Technology Co., Ltd.) weighing 200-220g were randomly divided into three groups of 10 rats each. The experimental animals were administered the drug once by gavage at a dose of 200mg / kg.

[0181] Control group: Curcumin suspension administered by gavage;

[0182] Experimental group 1: The curcumin nanocrystal composition of Example 4 was administered by gavage;

[0183] Experimental group 2: Gavage administration of curcumin nanocrystal suspension from Example 2.

[0184] Before the experiment, the experimental animals were fasted for 12 hours. At 6 min, 15 min, 0.5 h, 1 h, 1.5 h, 2 h, 3 h, 4 h, 8 h, 12 h, and 24 h after drug administration, approximately 0.5 mL of blood was collected from the orbital sinus. The blood samples were placed in anticoagulant tubes and centrifuged (8000 r / min * 10 min), with 200 μL of the supernatant precisely aspirated. The high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) method of this invention was used to detect the curcumin content in the plasma. Results are shown below. Figure 8 .

[0185] The above description of specific embodiments of the present invention does not limit the present invention. Those skilled in the art can make various changes or modifications based on the present invention, and as long as they do not depart from the spirit of the present invention, they should all fall within the scope of protection of the claims of the present invention.

Claims

1. A curcumin nanocrystal composition, wherein the curcumin nanocrystal composition is prepared from a curcumin nanocrystal suspension and a piperine suspension, wherein, The mass ratio of curcumin to piperine is 5-15:1, preferably 8-12:1, and more preferably 10:

1.

2. The composition of claim 1, wherein the curcumin nanocrystal suspension contains 20-35% curcumin nanocrystals, 1-10% steric protectant, and 1-10% charge protectant, wherein... The average particle size of curcumin nanocrystals is 200-320 nm.

3. The composition according to any one of claims 1-2, wherein the average particle size of curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

4. The composition according to any one of claims 1-3, wherein the stereoprotective agent is selected from any one or a combination of poloxamer, Tween, polyvinylpyrrolidone (PVP), poloxamer 108, poloxamer 407, poloxamer 124, and poloxamer 188.

5. The composition according to any one of claims 1-4, wherein the charge protection agent is selected from any one or a combination of sodium dodecyl sulfate (SDS), sodium deoxycholate, and sodium docusate.

6. The composition according to any one of claims 1-5, wherein, by weight-volume percentage, the curcumin nanocrystal suspension contains 20-35% curcumin nanocrystals, 1-10% poloxamer 407, and 1-10% sodium dodecyl sulfate, wherein, The nanocrystals of curcumin are 220-300nm, preferably 250-280nm.

7. The composition according to any one of claims 1-6, wherein, by weight-volume percentage, the curcumin nanocrystal suspension contains 30% curcumin nanocrystals, 5-9% poloxamer 407, and 1.5-5% sodium dodecyl sulfate, wherein, The average particle size of curcumin nanocrystals is 220-300 nm, preferably 250-280 nm.

8. The composition according to any one of claims 1-7, wherein the preparation of curcumin nanocrystal suspension comprises the following steps: dissolving the required amount of stereoprotectant and charge protectant in water, adding the required amount of curcumin, controlling the injection speed at 150-300 g / min, and grinding it in a grinder ((2000-3500 rpm) * (15-60 min)) to obtain curcumin nanocrystal suspension.

9. The method for preparing the curcumin nanocrystal composition according to any one of claims 1-8, wherein the curcumin nanocrystal composition is prepared from a curcumin nanocrystal suspension and a piperine suspension, wherein, The mass ratio of curcumin to piperine is 5-15:

1. The curcumin nanocrystal composition is prepared by mixing curcumin nanocrystal suspension and piperine suspension at a mass ratio of curcumin to piperine of 5-15:

1.

10. The use of the curcumin nanocrystal composition according to any one of claims 1-8 in the preparation of a medicament for preventing and treating exertional heatstroke and its resulting multi-organ damage or complications.

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