KR102573152B1 - Solubilisate with curcumin and optionally at least one other active substance - Google Patents

Abstract

In order to make available the health-promoting and healing properties of curcumin to the human or animal organism, in combination with at least one additional active substance, the present invention contains no more than 10% by weight, preferably no more than 7.5% by weight, most preferably no more than 7.5% by weight. at least one emulsifier having a curcumin content of 6% by weight and an HLB value of less than 18, preferably between 13 and 18, namely polysorbate 80 or polysorbate 20 or polysorbate 20 with polysorbate 80 A solubilize consisting of or containing a mixture is provided, wherein the curcumin-loaded micelles in the solubilize have an average diameter of 5 nm to 40 nm, preferably 6 nm to 20 nm, most preferably 7 nm to 10 nm And, the solubilizate is particularly intended for use as a pharmaceutical and/or dietary supplement for the treatment and/or prevention of inflammatory, cancer-related and other diseases.

Description

Solubilisates with curcumin and optionally at least one other active substance

The present invention relates to a solubilizate for use according to claim 1 comprising curcumin and optionally at least one further active substance. The present invention also relates to fluids containing such solubilisates, capsules filled with such solubilisates or fluids, and dietary supplements and/or pharmaceuticals containing such solubilisates.

Curcumin is being discussed as an active substance based on various potential pharmacological properties. For example, curcumin has been shown to have anti-inflammatory effects as well as efficacy against viruses and bacteria as well as against cancer, along with its use as an antioxidant. Thus, curcumin's uses include, for example, Parkinson's disease, Alzheimer's disease, diabetes, colon tumors, pancreatic cancer, and liver disorders.

In order for an active substance to enter the bloodstream after oral ingestion, the active substance must pass through the vascular barrier of the small intestine and then be metabolized in the liver to enter the hepatic vein as a bioavailable fraction. The remainder of the total active substance released into the body after ingestion is broken down by microorganisms in the intestine or eliminated in the feces or bile.

The present inventors have already created curcumin solubilizers with significantly increased bioavailability compared to natural curcumin. This soluble cargo is described in international patent application WO 2014094921 A1. Surprisingly, in some studies, in addition to its high bioavailability, certain formulations of this curcumin solubilizate have been found to have an unexpectedly greater impact, particularly in the reduction of inflammation or disease symptoms related to cancer.

Toxicity due to micellization of the active substance according to this invention compared to its native form can be ruled out based on the study of the MTT assay for cell viability. Verification of cell viability by MTT assay was performed by converting 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), a yellow water-soluble dye, into blue-violet water-insoluble formazan. based on reduction

The inventors have therefore set themselves the goal of providing a formulation that provides the health-promoting or therapeutic properties of curcumin usable to the human or animal organism by considering its combination with at least one additional active substance. In particular, it is an object of the present invention to provide the highest possible bioavailability of curcumin in combination with at least one further active substance.

This object is achieved in a surprisingly simple manner using the solubilizate according to claim 1 . This solubilizate comprises curcumin in an amount of less than 10% by weight, preferably less than 7.5% by weight, most preferably less than 6% by weight, and at least one emulsifier having an HLB value in the range of 13 to 18, i.e. polysorbate 80 or consisting of or containing polysorbate 20 or a mixture of polysorbate 20 and polysorbate 80, wherein the average diameter of the curcumin-loaded micelles is between 5 nm and 40 nm, preferably between 6 nm and 20 nm, most preferably between 7 nm and 10 nm, particularly associated with inflammation, cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, increased blood sugar, diabetes, metabolic syndrome, and/or autoimmune diseases, multiple sclerosis (MS) treatment and/or prevention of disease, reduction of visceral fat, thermogenesis, reduction of cholesterol, in particular reduction of LDL cholesterol, and/or reduction of blood glucose (blood sugar) and/or reduction of blood triglycerides, improvement of macular pigment density, reduction of oxidative stress and/or or for the reduction of fat accumulation in hepatocytes, in particular for the treatment and/or prevention of fatty liver disease, Friedreich's ataxia, lysosomal diseases, in particular Tay-Sachs disease, atherosclerosis, heart disease, arthritis for use as

The inventors have also found that curcumin's positive effects on the healing process or health maintenance can surprisingly be exploited synergistically, at least in part, in combination with other active substances. In one advantageous embodiment, the present invention provides curcumin in an amount of less than or equal to 10% by weight, preferably less than or equal to 8% by weight, most preferably from 3% to 7% by weight; and at least one emulsifier having an HLB value of less than 18, preferably in the range of 13 to 18, in particular polysorbate 80 or polysorbate 20 or a mixture of polysorbate 20 and polysorbate 80, or these It provides a soluble cargo containing

In one advantageous embodiment, the solubilizate is provided in orally administrable form, in particular in a dose of curcumin ranging from 0.5 mg/kg body weight to 1 mg/kg body weight, preferably 0.81 mg/kg body weight once daily.

For the purposes of this application, the term "active substance" refers to a substance that is provided in a pharmacologically effective concentration and is preferably added for the purpose of pharmacological effect. Here, the name of each active substance is understood to include substances that are converted in the body to the active substance and/or to its biologically active form.

For purposes of this application, such "active substances" include secondary phytochemicals produced by plants as compounds neither in energy metabolism nor in anabolic or catabolic metabolism. One group of secondary phytochemicals and thus active substances in the meaning of the present application are flavonoids. The active substance in the meaning of the present application also includes natural polyphenols such as resveratrol or polyphenols of licorice and natural phenols, especially chalcones such as xanthohumol, Plant extracts, which are substances extracted from parts of plants, are also included. These include extracts from the root material of hops or licorice plants. The active substance is referred to as an “extract” even though it is still dissolved in the extractant.

Active substances in the meaning of the present application also include enzymes. An example of an enzyme as an active substance in this application is serrapeptase. However, this application is not limited to this enzyme.

The resin extract of the frankincense tree, Boswellia serrata extract, contains several pentacyclic triterpenes, which are often referred to collectively as total boswellic acids ("total BAs"). The term "boswellic acids" refers to a group of chemical compounds that occur naturally in the resins of the aforementioned Frankincense tree. The two basic structures are α-boswellic acid and β-boswellic acid. Also, some derivatives of boswellic acid are known, especially compounds having a keto group at the 11-position and/or acetylated at the 3-position. Boswellic acids currently considered to be particularly significant in terms of pharmacological effects include α-boswellic acid (αBA) and β-boswellic acid (βBA) and their derivative 11-keto-β-boswellic acid (KBA); CAS 17019-92-0) and 3-O-acetyl-11-keto-β-boswellic acid (AKBA); CAS 67416-16-9), and 3-O-acetyl-α-boswellic acid (AαBA), and 3-O-acetyl-β-boswellic acid (AβBA). In particular, the derivative AKBA is believed to have an anti-inflammatory effect.

In the context of the present application, the term "Boswellia", in particular in the term "Boswellia solubles", means that the term "Boswellia" is an active substance derived from the resin of the Frankincense tree, i.e. at least one species of boswellic acid and/or at least one derivative of boswellic acid. The term "boswellic acid solubilisate" refers to a micellar formulation of at least one boswellic acid, which may also contain at least one boswellic acid derivative.

Xanthohumol (xanthohumol) is a flavonoid that occurs naturally in hops. It is a prenylated plant polyphenol assigned to chalcones and so far identified only in hops. Bitter hop varieties have a much higher content of xanthohumol than aromatic varieties. In tests, xanthohumol was found to be effective against the emergence and development of cancer cells. In laboratory experiments, it has been found that xanthohumol can protect nerve cells in the brain, which can help slow the progression of diseases such as Alzheimer's or Parkinson's.

Licorice flavonoid oil (LFO), composed of hydrophobic licorice polyphenols of medium-chain triglycerides, has a weight-reducing effect associated with body fat loss. In addition, antioxidant properties are attributed to the ethanol extract of licorice. The brand name of licorice flavonoid oil with a glabridin concentration of 3% is "KANEKA glavonoids ^TM ". Hereinafter, it is also referred to as "glavonoid".

Resveratrol is a phytoalexin with antioxidant properties belonging to polyphenols. This substance is also found in relatively high amounts in the skins of grapes, eg red grapes, but is also found in raspberries, mulberries, plums, peanuts and Japanese Knotweed. Resveratrol can also be isolated from the vine itself. According to an entry in the online encyclopedia Wikipedia, in vitro studies have shown potential anticancer activity and evidence of beneficial effects in diseases such as atherosclerosis, heart disease, Alzheimer's disease, arthritis and some autoimmune diseases.

Serratia peptidase or serrapeptase is a proteolytic enzyme produced by Serratio, a bacterium that lives in the intestines of silkworms. Serrapeptase is known to have beneficial effects in relieving pain, inflammation, traumatic edema and excessive mucus secretion by the organism. It is said to be effective as an anti-inflammatory and analgesic similar to acetylsalicylic acid, ibuprofen or other non-steroidal pain relievers. It is also said to induce fibrinolytic anti-inflammatory and anti-edema actions in tissues. Like all enzymes, serrapeptase is sensitive to acids produced by the stomach. Accordingly, it is an object of the present invention to provide a formulation that permits gastrointestinal passage.

The solubilizate according to the present invention contains at least one boswellic acid and/or at least one boswellic acid derivative in an amount of 10% by weight or less, preferably 8% by weight or less, most preferably 4.7% to 6.6% by weight. can do.

Due to the high Boswellia ratio, the present invention, in one advantageous embodiment thereof, is a source of at least one boswellic acid and/or at least one boswellic acid derivative by extraction with ethyl acetate. serrata ) resin, wherein boswellic acid is contained in a concentration of at least 85% by weight of the extract.

The solubilizer according to the present invention may contain xanthohumol in an amount of 10% by weight or less, preferably in an amount of 5% by weight or less, most preferably in an amount of 1 to 3% by weight.

Due to the high proportion of xanthohumol, the present invention, in one advantageous embodiment thereof, contains as a source of xanthohumol an ethanolic extract of hard resin from hops, wherein the concentration of xanthohumol in said extract is from 65% to 95% by weight % range, preferably a concentration range of 80% to 92% by weight of the solubilizate is envisaged. In particular, "Xantho-Flav Pure" or "Xantho-Flav", which will be discussed in more detail below, can be used as the source of xanthohumol in the present invention.

The solubilizate according to the present invention contains licorice root extract, in particular a fluid containing a hydrophobic solution of licorice root extract, preferably a glavonoid, and/or glabridin, in an amount of not more than 35% by weight, preferably not more than 20% by weight. , most preferably in an amount of 0.3% to 17% by weight.

The solubilizer according to the present invention may contain resveratrol in the range of 1% to 15% by weight, most preferably in the range of 5% to 10% by weight.

The solubilizate according to the present invention may contain at most 3% by weight of serrapeptase, preferably in the range of 0.1% to 2% by weight, most preferably in the range of 0.18% to 0.35% by weight.

The solubilizate according to the present invention may contain coenzyme Q ₁₀ up to 10% by weight, preferably in the range of 0.1% to 5% by weight, most preferably in the range of 0.5% to 1.5% by weight.

The solubilizate according to the present invention contains α-liponic acid at most 10% by weight, preferably in the range of 0.1% to 5% by weight, most preferably in the range of 0.8% to 2.5% by weight. may contain

Solubleates consisting of or containing curcumin and at least one further active substance are also in the context of the present invention as antibiotics and/or for the treatment of inflammation, cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, increased blood sugar, Treatment and/or prevention of diseases associated with diabetes, metabolic syndrome, and/or autoimmune diseases, multiple sclerosis (MS), reduction of visceral fat, thermogenesis, reduction of cholesterol, particularly LDL cholesterol, and/or blood glucose (blood sugar) medicines for reducing and/or reducing blood triglycerides, improving macular pigment density, reducing oxidative stress and/or reducing fat accumulation in hepatocytes, in particular fatty liver disease, Friedreich's ataxia, lysosomal diseases, especially Tay-Sachs disease disease), arteriosclerosis, heart disease, arthritis, can be advantageously provided or used for use as a pharmaceutical for the treatment and/or prevention.

In particular, the present invention relates to the use of the aforementioned solubilizers as anti-inflammatory drugs and/or as antibiotics and/or for cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, increased blood sugar, diabetes, metabolic syndrome, and/or autoimmune diseases, As a medicine for multiple sclerosis (MS), visceral fat reduction, fever, cholesterol reduction, in particular LDL cholesterol reduction, and/or blood glucose (blood sugar) reducing effect and/or blood triglyceride reducing effect, macular pigment density improving effect, oxidation As a medicine with a stress-reducing effect and/or a fat accumulation reducing effect in hepatocytes, especially for fatty liver, Friedreich's ataxia, lysosomal diseases, especially Tay-Sachs disease, arteriosclerosis, heart disease, and arthritis It is provided to be used as a medicine with

In this context, the solubilizate according to the present invention also has a total curcuminoid concentration in human plasma measured 1 hour after oral administration of 500 mg of curcumin in the form of the above-mentioned solubilizer of about 500 ng curcuminoid per mL plasma ± 100 ng curcuminoids per mL plasma has proven beneficial. Total curcuminoid concentrations in human plasma, measured as the area under the total curcumin plasma concentration versus time curve (AUC) over 24 hours, ranged from about 9,500 to about 10,000 nmol h/L. Thus, the solubilizate is available in large quantities in the body even after oral administration.

The present invention is advantageous because it provides a solubilizer having very good anti-inflammatory properties. The anti-inflammatory activity measured as the concentration of C-reactive protein (CRP) in the serum of arthritic rats after oral administration of the solubilizate according to the present invention at a dose of 5 mg/kg body weight of curcumin was about 2100 pg /mL to about 2500 pg/mL, and the concentration of CRP after one-time administration of the solubilizate at a dose of 10 mg/kg body weight of curcumin is in the range of about 1400 pg/mL to about 1800 pg/mL.

The anti-inflammatory effect measured as the concentration of myeloperoxidase (MPO) in the serum of arthritic rats after one administration of the solubilizate according to the present invention at a dose of 5 mg/kg body weight of curcumin was about 800 mU/mL to about 900 mU /mL range. These values are much lower than those of natural curcumin, as detailed below.

Curcumin and boswell, measured as concentrations of C-reactive protein (CRP) in the serum of arthritic rats after one-time administration of the solubilizate according to the present invention at a dose of 5 mg/kg body weight of curcumin and 10 mg/kg body weight of boswellic acid The anti-inflammatory activity of the solubilizer of the present invention containing Wellia is about 1200 pg/mL, compared to about 3200 pg/mL to about 3500 pg/mL after administration of natural curcumin and Boswellia at the same dose, respectively. to about 1500 pg/mL.

Curcumin and boswell, measured as concentrations of myeloperoxidase (MPO) in the serum of arthritic rats after one administration of the solubilizate according to the present invention at a dose of 5 mg/kg body weight of curcumin and 10 mg/kg body weight of boswellic acid Since the anti-inflammatory effect of the solubilizate of the present invention including Wellia is about 750 mU/mL to about 815 mU/mL, therefore, the value after administering natural curcumin and Boswellia at the same dose, respectively, is about 1150 mU/mL to about 1250 mU/mL, which is much lower.

Enzymatic units (U) have been replaced by kat to indicate enzymatic activity. Enzymatic units (U) continue to be used in medicine and clinical chemistry because their numerical value changes when katal is used. One U of enzyme unit corresponds to 1 micro-mole of substrate conversion per minute.

The present invention also relates to use in agriculture, fish farming and/or horticulture and/or food hygiene and/or use as a disinfectant and/or packaging, preferably beef, poultry or fish packaging and/or use as a disinfectant. It is advantageous because it can be provided for

In particular, in the case of a solubilisate containing curcumin as the only active substance, it could be shown that treatment with the solubilisate according to the present invention resulted in a significant reduction in the microbial load in terms of bacterial contamination of food. In this case, curcumin is effective as a photosensitizer. Applied topically to a surface infected by a microorganism, which is absorbed by the microorganism on that surface, the surface is exposed to light to activate the photosensitizer, thereby generating reactive oxygen species ("oxygen radicals") in the microorganism Microorganisms are thereby killed.

In the field of food, for example, in the field of antimicrobial packaging, food such as meat is sprayed with an aqueous solution of the soluble cargo according to the present invention, and then packaged with a transparent film made of PP, PE, PC, etc., and then exposed to light, Food surfaces can be kept aseptic. Corresponding applications for aseptic packaging of other items are also included within the scope of the present invention. Examples thereof include packaging of surgical instruments and similar instruments.

An advantage of the formulation as a solubilizate according to the invention for this type of application is that the active substance, in particular curcumin, can be used in aqueous dilution in very high multiples, thereby virtually completely eliminating the color load of the treated surface. that it can Due to the large number of micelles of very small size, a homogeneous and sufficient distribution of the active substance of the solubilizate on the surface is achieved even in the case of a high-fold dilution treatment. Thus, for example, the present invention is applicable in the field of packaging in a simple and very cost-effective manner and in the most aseptic manner possible.

In order to provide stable micelles of the active substance, the emulsifier content, in particular the polysorbate content, depends on the further components contained in the solubilizer, but in the context of the present invention is at least 70% by weight, preferably between 75% and 95% by weight % range, most preferably from 79% to 88% by weight.

Depending on the particular application, the solubilizate of the present invention may contain, for example, at most 20% by weight, preferably at most 15% by weight of ethanol, and/or at most 25% by weight, preferably at most 12% to 20% by weight, most preferably up to 10% by weight of glycerol, and/or additionally up to 10% by weight of water, preferably up to 7% by weight of water. With the addition of ethanol, the content of polysorbate can be reduced, which is advantageous in view of the ADI value of polysorbate (25 mg/kg body weight) recommended by WHO. The polysorbate content can also be reduced by adding glycerol.

The solubilizate of the present invention exhibits a narrow particle size distribution with a small average particle size even under the physiological condition of gastrointestinal passage; The diameter distribution of the micelles in the solubilizer dilution at pH 1.1 and 37° C. at a ratio of 1:500 with distilled water is about d ₁₀ = 6 nm to about d ₉₀ = 20 nm. These values are obtained from the volume distribution. Detailed particle size analysis of the micelles of the solubilizate will be described later.

Evidence of improved bioavailability (of the solubilisate of the present invention) compared to a composition or curcumin composition of unmicellated curcumin and at least one additional active substance according to the present invention is based on a much more readily accessible measurement technique, the It can be obtained by measuring turbidity. As a result of the formulations according to the present invention, the turbidity of the solubilisate is 1:50 or 1:500 diluted with water under physiological conditions (pH 1.1 and 37° C.) according to the ISO 7027 standard specification for dilutions of the solubilisate. When measured by light scattering measurements using infrared light, it is preferably less than 25 FNU, more preferably less than 3 FNU.

In order to facilitate oral application of the solubilisate of the present invention in a simpler and more convenient manner from the point of view of consumers or patients, the present invention also provides capsules filled with the above-described solubilisate, wherein the capsule is a soft gelatin capsule or a hard in the form of gelatin capsules or soft gelatin-free capsules or hard gelatin-free capsules, such as cellulose capsules.

Also, in the context of the present invention, the solubilizate according to the present invention can be incorporated into other fluids, particularly liquids. Small micelles filled with active substances are retained even in this case. Accordingly, the present invention also provides a fluid containing a solubilizate as described above, wherein the fluid is selected from the group consisting of foods, dietary supplements, beverages, cosmetics and pharmaceuticals. In the context of the present invention, the fluid may in particular include an aqueous dilution of the solubilizate.

The present invention also relates to the treatment and/or prevention of diseases associated with inflammation, cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, elevated blood sugar, diabetes, metabolic syndrome, and/or autoimmune diseases, multiple sclerosis (MS), reducing visceral fat, thermogenesis, reducing cholesterol, especially LDL cholesterol, and/or reducing blood glucose (blood sugar) and/or reducing blood triglycerides, improving macular pigment density, reducing oxidative stress, and/or reducing fat accumulation in hepatocytes. for the treatment and/or prophylaxis of, in particular, fatty liver disease, Friedreich's ataxia, lysosomal diseases, in particular Tay-Sachs disease, arteriosclerosis, heart disease, arthritis, diseases associated with improvement of macular pigment density , wherein the method comprises administering to the dietary supplement consumer or patient a solubilisate according to the present invention, in particular as a capsule or as a fluid, in particular by the oral route, in particular once daily.

In a preferred embodiment of the method of the present invention, the solubilizate is administered to the dietary supplement consumer or patient at a dose ranging from 0.5 mg/kg body weight to 1 mg/kg body weight of curcumin, preferably at a dose of 0.81 mg/kg body weight. .

In a preferred embodiment of the method of the present invention, the solubilizate is administered to the dietary supplement consumer or patient with Boswellia at a dose ranging from 1 mg/kg body weight to 2 mg/kg body weight, preferably at a dose of 1.62 mg/kg body weight. dosing

In a preferred embodiment of the method of the present invention, the solubilizate is administered to the dietary supplement consumer or patient at a dose ranging from 0.5 mg/kg body weight to 1 　mg/kg body weight of xanthohumol, preferably 0.81 mg/kg body weight. do.

For the preparation of a solubilisate according to the present invention comprising curcumin and at least one further active substance, the prepared solubilates are either individually mixed together or the solubilisate containing curcumin and at least one further active substance is directly mixed. can be manufactured

The present invention also provides a method for preparing the above-mentioned solubilisate. If co-micellarization of curcumin and at least one further active substance is desired, the present invention comprises the following steps:

(a) providing polysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20 and polysorbate 80;

(b) adding at least one further active substance, in particular Boswellia serrata extract and/or xanthohumol;

(c) adding curcumin powder;

Including, providing a first modified manufacturing method,

wherein step (a) comprises heating to a temperature in the range of 40°C to 62°C, preferably in the range of 45°C to 57°C, most preferably in the range of 48°C to 52°C; and

Step (b) maintains the temperature unchanged compared to step a), or in the temperature range of 60°C to 75°C, preferably in the temperature range of 61°C to 70°C, most preferably in the range of 63°C to 67°C. including heating to a temperature range; and

Step (c) comprises heating to a temperature range of 82°C to 97°C, preferably to a temperature range of 83°C to 92°C, most preferably to a temperature range of 85°C to 89°C.

By this method of preparation, it is possible to prepare a solubilizate capable of forming micelles loaded with curcumin and at least one additional active substance in aqueous dilution. For this purpose, it is possible to intermix at least two active substances under suitably adapted temperature control and then add them as a mixture, in combined form.

Specifically, prior to step b), step b1) of adding water at a temperature in the range of 40°C to 62°C, preferably in the range of 45°C to 57°C, most preferably in the range of 52°C is performed. can

Additionally or alternatively, step b1) comprises adding ethanol at a temperature in the range of 40°C to 62°C, preferably in the range of 45°C to 57°C, most preferably in the range of 48°C to 52°C. may include

Another option for the manufacturing process involving co-micellization of curcumin and at least one further active substance is the following steps:

(a) polysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20 and polysorbate 80 and at least one first active substance, in particular α-lipoic acid, preparing a first formulation by providing;

(b) preparing a second formulation by providing water and at least one further active substance, in particular serrapeptase;

(c) adding at least one further active substance, in particular xanthohumol and/or Boswellia serrata extract and/or glavonoids and/or resveratrol and/or coenzyme Q ₁₀ , and curcumin powder to the second formulation from step (b);

(d) combining the first formulation from step (a) and the second formulation from step c), wherein the temperature is between 18° C. and 22° C. during steps (a) to (d). ;

(e) heating to a temperature range of 80° C. to 97° C., preferably to a temperature range of 83° C. to 92° C., most preferably to a temperature range of 85° C. to 89° C.

It is provided by a second variant method of the present invention by a method comprising

Step c) may further comprise adding ethanol and/or glycerol and/or MCT oil and/or polysorbate 20 and/or polysorbate 80 and/or a mixture of polysorbate 20 and polysorbate 80. can

Another option for the production process involving co-micelletization of curcumin and at least one further active substance is the following steps:

(a) providing a mixture of polysorbate 80 and/or polysorbate 20 and/or polysorbate 20 and polysorbate 80 and a mixture of glycerol and ethanol;

(b) adding at least one further active substance, in particular an ethanolic extract of the hard resin from hops, in particular Xantho-Flav Pure powder,

step (a) comprises heating to a temperature range of 40°C to 62°C, preferably to a temperature range of 45°C to 57°C, most preferably to a temperature range of 48°C to 52°C;

wherein step (b) comprises heating to a temperature range of 60°C to 75°C, preferably to a temperature range of 61°C to 70°C, most preferably to a temperature range of 63°C to 67°C; and

(c) adding curcumin powder at a temperature ranging from 70°C to 92°C, preferably from 75°C to 87°C, most preferably from 78°C to 82°C;

(d) adding glavonoids while heating to a temperature range of 80° C. to 97° C., preferably 83° C. to 92° C., most preferably 85° C. to 89° C.

It is provided by a third variant method of the present invention by a method comprising

The present invention also relates to solubilisates which, at least immediately after their preparation, exhibit micelles in an aqueous dilution loaded with curcumin alone or otherwise with another active substance alone. Accordingly, the present invention also provides a process for preparing a solubilisate as described above by mixing a curcumin solubilisate and a solubilisate of one further active substance, in particular in a quantitative ratio of individual solubilisates of 1:1.

The present invention will be described in more detail with reference to specific examples below. The following ingredients were used:

curcumin

Green Leaf Extraction Pvt Ltd. (Kerala, India) product code EP-5001, trade name "Turmeric Oleoresin Curcumin Powder 95% (Turmeric Oleoresin Curcumin Powder 95%)" as curcumin used This curcumin powder has CAS Number 458-37-7. It is a natural product obtained by solvent extraction of the rhizomes of Curcuma Longa . This powdered curcumin content is at least 95% according to manufacturer specifications. This curcumin content was determined by ASTA method 18.0.

Nilam Phyto-Extracts (Mumbai, India), used as curcumin in the illustrative embodiments described below, as an example of a replacement for the aforementioned “Oleorescein Turmeric 95%” curcumin powder from Green Leaf Extraction Pivti Ltd. 95% Curcumin Extract, or Curcumin BCM-95-SG or Curcumin BCM-95-CG from Eurochem GmbH (Grbenzel, Germany), or Curcuma Oleoresin 95% from Henri Lamotte Oils (Bremen, Germany) (Curcuma Oleoresin 95%).

boswellia

In the context of the present invention, the term “Boswellia” refers in particular to an extract derived from the resin of the Frankincense plant. Specifically, an extract of the Boswellia serrata species was used, which was obtained from the resin of a plant with the scientific name Boswellia serrata by extraction with ethyl acetate, and was used as Product code "HC22519" of N.V. (Frutarom Belgium NV, Ronderzel, Belgium). Solubilisates containing this extract are also referred to as "boswellic acid solubilisates" due to their boswellic acid content.

Besides extracts from resins of the Frankincense plant, it is also possible to use boswellic acids and/or derivatives of boswellic acids for the purposes of the solubilisates according to the invention. In particular, the following may be considered; Alpha-boswellic acid (CAS number 471-66-9), beta-boswellic acid (CAS number 631-69-6) and their derivatives, 3-O-acetyl-alpha-boswellic acid (CAS number 89913-60-0 ), 3-O-acetyl-beta-boswellic acid (CAS number 5968-70-7), 11-keto-beta-boswellic acid (KBA, CAS number 17019-92-0), and 3-O-acetyl-11 -keto-beta-boswellic acid (AKBA, CAS number 67416-61-9).

Xanthohumol

Simon H. Use of "Xantho-Flav" or "Xantho-Flav Pure", a product of the "Hopsteiner" brand of Simon H. Steiner, Hopfen, GmbH (Mainburg, Germany), as a source of xanthohumol did Both of these are natural products produced from hops. The active substance is xanthohumol, a polyphenol of hops. These are yellow powders with a xanthohumol content of 65% and 85% for "Xantho-Flav" and at least 85% for "Xantho-Flav Pure", according to the manufacturer's specifications.

The concentration of xanthohumol and isoxanthohumol in "Xantho-Flav Pure" is quantified by the manufacturer according to UV spectrophotometric analysis or HPLC EBC 7.8 using an external calibration standard pure XN (370 nm) or IX (290 nm). “Xantho-Flav Pure” contains a very high concentration of the prenylated flavonoid xanthohumol. In an exemplary embodiment in the context of the present application, batch number 9432 of "Xantho-Flav Pure" was used.

glavonoids/glabridins

"Glavonoid" is the product name of a composition from Kaneka Corporation (Osaka, Japan) that contains glabridin as an active substance. Glabridin is a flavonoid from the licorice plant ( Glycyrrhiza glabra ). The product “Kaneka Glavonoid” contains, according to the manufacturer, 30% licorice extract and 70% edible oil. "Kaneka glavonoids," according to the manufacturer, are standardized to 3% glabridin, the main polyphenol in the licorice plant. The CAS number for glabridin is 59870-68-7.

resveratrol

The product name “eveResveratrol” was used as resveratrol. This is transresveratrol obtained by fermentation. This product is provided as a white powder without additives. The content of transresveratrol is at least 98% by weight with the balance being water. The CAS number for this product is 501-36-0 and the product code is RSXOL 9800.

Serrapeptase

Shaanxi Pioneer Biotech Co. Ltd's product name Serrathiopeptidase (Batch number PBD 20170708) was used as the serrapeptase. It is an off-white to light brown powder.

coenzyme Q ₁₀

Coenzyme Q ₁₀ was purchased from Xiamen Kingdomway Group Company. It is prepared by microbial fermentation and contains less than 0.5% of the cis isomer according to the manufacturer's specifications.

α-Liponic acid (alpha-liponic acid)

Alpha-liponic acid was purchased from Jiangsu Tohope Pharmaceutical Co. Ltd. (China).

Polysorbate 80

The source of polysorbate 80 was the material "TEGO SMO 80 V FOOD" with specification code "K04 EU-FOOD" from Evonik Nutrition & Care GmbH (Essen, Germany). This product meets EU requirements for food additives E 433. As a substitute for Evonik's TEGO SMO 80 V described above as polysorbate 80 in an exemplary embodiment described below, TEGO SMO 80 V from InCoPA GmbH (Ilerthyssen, Germany), or Croda GmbH ( Crillet 4/Tween 80-LQ-(SG) from CRODA GmbH, Nettetal, Germany, or Lamesorb SMO 20 from Univar or Kolb Distributions AG, Heidingen, Switzerland, and Cotilen-O/1 VL (Kotilen-O/1 VL) can be used.

polysorbate 20

The source of polysorbate 20 was the material "TEGO SML 20 V FOOD" with specification code "K09 EU-FOOD" from Evonik Nutrition & Care GmbH (Essen, Germany). This product meets EU requirements for food additives E 433. As polysorbate 20 in the context of the present invention, as a substitute for the aforementioned TEGO SML 20 from Evonik, Crillet 1/Tween 20-LQ-(SG) from CRODA GmbH (Nettetal, Germany) can be used. there is.

ethanol

In the context of the present invention, ethanol was purchased from Berkel Pfalzische Spritfabrik GmbH & Co. KG. According to the specification, in the case of "Undenatured Neutral Alcohol 1411U Customs", the ethanol content of this product is about 92.6 to 95.2% by weight.

glycerol

The product used as glycerol in the context of the present invention was “Glycamed 99.7%.” According to the manufacturer's specifications the glycerol content of this product is at least 99.5%.

medium chain triglycerides

Medium chain triglycerides (MCTs) are triglycerides containing medium chain fatty acids. Medium chain fatty acids include caproic, caprylic, capric and lauric acids. These are saturated fatty acids that occur naturally in tropical vegetable fats such as, for example, coconut oil and palm kernel oil. These are also contained in milk fat to a small extent. However, pure MCT oil does not exist in nature, and pure MCT oil can be obtained synthetically. Individual MCTs or mixtures of different MCTs may be used as medium chain triglycerides within the scope of the present invention. Medium-chain triglycerides are available in the form of MCT oil Delios VK Kosher (from Cognis GmbH, Monheim, Germany) or as MCT oil (70/30) (Rofetan GTCC 70/30) (from DHW Deutsche Hydrierwerke Rodleben GmbH, Dessau-Roslau, Germany). , CAS number 73-398-61-5).

Medium chain triglycerides can also be used in the form of the product ROFETAN DTCC 70/30 (Ph. Eur.). It is a caprylic/capric triglyceride with CAS number 73398-61-5. This product corresponds to the monograph "Medium Chain Triglycerides" in the European Pharmacopoeia in force at the date of filing. The manufacturer is Ecogreen Oleochemicals DHW (Deutsche Hydrierwerke GmbH, Rodleben, Germany).

mixed tocopherols

For example, as mixed tocopherols, 70% mixed tocopherols in vegetable oil Vitapherole T-70 Non GMO, manufactured by VitaeNaturals, can be used (E306, CAS number 59-02-9, 16698-35 -4, 54-28-4, and 119-13-1).

When water is added in preparing the solubilize, distilled water is used.

Dr. Faculty of Pharmacy, Cairo University. Ing. M.T. Professor Khayyal studied the anti-inflammatory effects of curcumin, a combination of curcumin and boswellia, or a combination of curcumin and xanthohumol, in each case the natural form and the solubilized form for the present invention.

Efficacy as an anti-inflammatory marker and antioxidant was determined. Female Wistar rats weighing 150 to 200 g were tested by Pearson et al. (1956) were exposed to adjuvant-induced arthritis. On day 0, animals were administered 0.1 ml of Freund's adjuvant (FCA) by subplantar injection into the right hind paw. Animals were randomly divided into 12 groups of 8 animals each.

Group 1 was the control group.

Group 2 was administered diclofenac as a reference drug at a dose of 3 mg/kg body weight.

Group 3 was administered natural curcumin at a dose of 5 mg/kg body weight.

Group 4 was administered curcumin solubilized according to the present invention at a dose of 5 mg/kg body weight.

To group 5, natural curcumin was administered at a dose of 10 mg/kg body weight,

Group 6 was administered the same dose of solubilized curcumin.

Group 7 was administered natural xanthohumol at a dose of 5 mg/kg body weight,

Group 8 was administered the same dose of solubilized xanthohumol.

Group 9 was administered a mixture of natural curcumin at a dose of 5 mg/kg body weight and natural Boswellia extract at a dose of 10 mg/kg body weight.

Group 10 was administered a mixture of solubilized curcumin and solubilized Boswellia at equal doses respectively.

Group 11 was administered a mixture of natural curcumin at a dose of 5 mg/kg body weight and natural xanthofumol at a dose of 5 mg/kg body weight,

Group 12 was administered a mixture of solubilized curcumin and solubilized xanthohumol at equal doses respectively.

All extracts or solubilizers were orally administered once a day from day 0 to day 21 after vaccination with an adjuvant. After 21 days, the animals were killed and serum samples were prepared and stored at -80°C. Measured for myeloperoxidase (MPO), C-reactive protein (CRP), total antioxidant potency (TAC), and thiobarbituratic acid reactive substances (TBARS).

The experimental results are described below with reference to the accompanying drawings:
1A is a diagram illustrating the effects of native and solubilized forms of curcumin and diclofenac on serum CRP levels (pg/L);
Figure 1B compares the effect of native and solubilized forms of curcumin (5 mg/mL) on serum CRP levels (pg/L) with that of diclofenac when administered with boswellia or with xanthohumol;
Figure 2 illustrates the effects of native and solubilized forms of curcumin and diclofenac on serum MPO levels (mU/mL);
Figure 3 compares the effect of native and solubilized forms of curcumin on serum MPO levels (mU/mL) with that of diclofenac when administered with boswellia or with xanthohumol.

First, the effect on C-reactive protein (CRP) was studied. C-reactive protein is a specific marker for anti-inflammatory activity. Both native and solubilized forms of curcumin suppressed serum CRP levels in rats in a dose-dependent manner, but at selected doses the solubilized form was twice as effective as the native form and significantly more effective than diclofenac ( FIG. 1A ) .

Coadministration of native form of curcumin with Boswellia did not significantly change its inhibitory effect on serum CRP (FIG. 1B). However, when curcumin and boswellia were administered together in solubilized form and at appropriate doses, the anti-inflammatory effect was enhanced. In this case, Boswellia potentiated the effect of curcumin more than xanthohumol at the doses considered.

Myeloperoxidase (MPO) in plasma plays a central role as a pro-inflammatory mediator in rheumatoid arthritis and is an indicator of invasion of neutrophil granulocytes into affected tissues. Its concentration is elevated in patients with rheumatoid arthritis and causes oxidative stress. As a result of the study, the MPO concentration was effectively reduced by solubilized curcumin in the same manner at both doses considered, but there was no significant difference with diclofenac. However, natural curcumin had no effect on MPO levels (Figure 2). Concomitant administration of curcuman and boswellia in both native and solubilized forms did not improve the effectiveness of curcumin in lowering MPO levels.

Oxidative stress is one of the major factors contributing to joint destruction in rheumatoid arthritis (RA). Increased production of so-called “reactive oxygen species (ROS)” reduces the supply of endogenous antioxidants, ultimately leading to cell destruction. Neutrophil granulocytes released from rheumatoid joints generate free oxygen radicals, which increase the formation of lipid peroxides, which is manifested in an increase in serum TBARS. Therefore, an increase in antioxidant status represented by an increase in TAC can be used as an indication of protection against the development of degenerative inflammatory processes. There is an inverse relationship between TAC levels and levels of TBARS, with high levels of antioxidant capacity TAC corresponding to low TBARS concentrations.

The results of the studies conducted showed that curcumin in its natural form did not significantly affect TBARS or TAC levels at both doses considered. Solubilized curcumin according to the present invention reduced TBARS levels and increased TAC at all selected doses, with little difference to the effect of diclofenac.

These data are summarized in the table below. The table shows that when natural and solubilized forms of curcumin and boswellia, alone or in combination with diclofenac, were administered at a dose of 3 mg/kg body weight once daily for 21 days, in the serum of arthritic rats (n=8), Data on the antioxidant capacity TAC and its effect on the thiobarbituric acid reactive substance TBARS are described. Figures presented represent mean values ± standard error of the mean (SEM).

According to the results presented in the table, co-administration of curcumin and boswellia in their natural form did not show any significant effect in reducing oxidative stress. However, in solubilized form, these combinations were as effective as diclofenac in reducing TBARS and increasing TAC in the serum of arthritic rats.

Use of Curcumin Solublezate According to the Invention for Combating Cancer Cells Against Breast Cancer Cell Lines (MCF-7) and (T47D), Liver Cancer Cell Line (Hepg-2), Colon Cancer Cell Line (HCT-116), and Prostate Cancer Cell Line (PC3) As a result of the first study on , it shows very good results in that cells capable of surviving even after treatment are achieved at the lowest rate. Curcumin solubilization allowed for a reduction to a "surviving fraction" ranging from about 15% to about 25%.

Unless otherwise stated, the particle size of the micelles in the aqueous dilution of the solubilizate according to the present invention was measured according to the principle of dynamic light scattering using laser light with a wavelength of 780 nm. Particle size measurements were performed using a ParticleMetrix NANO-flex backscatter particle analyzer. The measurement principle is based on dynamic light scattering (DLS) in a 180° heterodyne backscattering setup.

To experimentally determine the turbidity of the solubilizate according to the present invention, the turbidimeter is calibrated with a standard suspension. Therefore, instead of the measured light intensity, the concentration of the calibration suspension is displayed. Thus, when any suspension is measured, the indicated value means that the respective liquid causes the same light scattering as the standard suspension at the indicated concentration. The internationally defined turbidity standard is formazine. The most common unit is the FNU designation, or Formazin Nephelometric Units. This is the unit used in water treatment, for example to measure at 90° in compliance with the requirements of the ISO 7072 standard.

In order to prepare a solubilizate according to the present invention comprising curcumin as an active substance and at least one additional active substance, individually prepared solubilisates are mixed with each other or curcumin and at least one additional active substance or several additional active substances A solubilizate containing the active substance can be prepared directly.

Curcumin Soluble Cargo

For illustrative purposes, a 7% curcumin solubilizate is prepared. for teeth,

925 g polysorbate 80 and

75g curcumin powder 95% (= 71.2g curcumin)

this is used

Heat the polysorbate 80 to 48-52°C. Curcumin powder is added to the polysorbate under agitation while further heating to a temperature in the range of 95-97°C. The powder is added at an appropriate rate so that it is drawn into the emulsifier homogeneously during stirring. After cooling to a temperature below 60° C., the curcumin solubilisate is bottled. This solubilisate was used to prepare curcumin and boswellia solubilisates.

When diluted 1:500 with water at pH 1.1 and a temperature of 37°C, 7% curcumin solubilize exhibits an average turbidity of 0.9 FNU.

However, it should be noted that the curcumin content can be further increased without having to accept adverse consequences, for example in terms of micelle stability. A composition consisting of 100 g 95% curcumin powder and 900 g polysorbate 80 is a product as stable as a composition consisting of 120 g 95% curcumin powder and 880 g polysorbate 80 or 70 g 95% curcumin powder and 930 g polysorbate 80 results in

Also, polysorbate 80 can be wholly or partially substituted for polysorbate 20. For example, to prepare a curcumin solubilizate with only polysorbate 20, 894 g of polysorbate 20 and 106 g of 95% curcumin powder can be used. Polysorbate 20 is heated to about 63°C to about 67°C. Slowly add the curcumin powder to the polysorbate 20 while stirring. Continue heating to about 83° C. to about 87° C. while adding the curcumin powder. The solubilized product obtained is slowly cooled to less than about 45° C. to prepare for bottling.

Otherwise, the preparation of these variants corresponds to that described above. Up to about 11% of solubilize can be prepared in this way.

1.5% serrapeptase solubilizate

The following are used:

15 g serrapeptase: 20,000 U/mg serrathiopeptidase = 300,000,000 U;

15g water,

16.5 g MCT oil,

953.5 g Polysorbate 80.

At a temperature ranging from 18 to 22° C., water is mixed with serrapeptase and the mixture is homogenized. This means that the serrapeptase is distributed as evenly as possible in the water. This creates conditions in which the serrapeptase is largely completely soluble in water. While heated to a temperature ranging from 83 to 87° C., the MCT oil is incorporated into the water-serrapeptase mixture with constant agitation. Stirring is performed sufficiently to dissolve the serrapeptase homogeneously in the water. At a constant temperature, polysorbate 80 is added under stirring and homogenized. Stirring is performed sufficiently to distribute the polysorbate 80 homogeneously. The product was cooled to a temperature below 60° C. and bottled. It is then stored in a dark room below 25°C.

300,000 U/g corresponds to 15 mg/g enzyme units of 1.5% serrapeptase. Upon dilution in water at a ratio of 1:50, the turbidity of this solubilizate was determined under physiological conditions of pH 1.1 and 37°C. The resulting value was 1.8 FNU.

For particle size analysis of serrapeptase solubilisate, this solubilis was first diluted with distilled water in a ratio of 1:500 and heated to 37° C. with constant stirring using a magnetic stirrer and hot plate. The pH was then adjusted to 1.1 using 32% hydrochloric acid. The sample was then measured immediately. The results are summarized in the table below and the average value of the two measurement data is obtained.

10% resveratrol solubles

The following was used:

100g resveratrol,

45g MCT Oil,

600g polysorbate 80,

180 g polysorbate 20, and

75 g mixed tocopherols.

At a temperature ranging from 18 to 22° C., the polysorbate, mixed tocopherols and MCT oil are mixed and homogenized with sufficient agitation to ensure a homogeneous distribution of these components. To a mixture or solution of polysorbate, MCT oil and mixed tocopherols, resveratrol is added while heating to a temperature in the range of 83 to 87° C. and with sufficient agitation, so that the resveratrol is absorbed homogeneously into the formulation containing the emulsifier. Once a homogeneous and transparent product is obtained, it is cooled to a temperature below 30° C. and bottled. The product is yellowish, transparent and viscous, stored in the dark at 25°C.

For therapeutic purposes, for a dose based on 3,500 mg of resveratrol raw material in its natural form, the following calculation is applied to the equivalent amount of solubilizate:

A fill of three capsules, each with 675 mg of solubilizer, corresponds to an amount of 2,025 mg or 200 mg of the active substances resveratrol and 100 mg of mixed tocopherols per day. For DV factor 1:17 the following applies: 200 mg x 17 = 3,400 mg of resveratrol pure substance.

The turbidity of this solubilizate was also measured under physiological conditions (pH 1.1; 37° C.) at a 1:50 dilution with water. The resulting value was 16.1 FNU.

5% Coenzyme Q ₁₀ fusible cargo

The following was used:

57.5 g coenzyme Q ₁₀ ,

160 g MCT oil, and

782.5 g Polysorbate 80.

The polysorbate is heated to a temperature ranging from 83°C to 87°C. Coenzyme Q ₁₀ powder is then incorporated under agitation. Stir vigorously enough to evenly distribute the ingredients. Coenzyme Q ₁₀ is dissolved in the polysorbate 80 while maintaining this temperature or reheating to a temperature in the range of 83-87°C. The MCT oil is then incorporated while maintaining the temperature. Once a homogeneous and transparent product is obtained, it is cooled to a temperature below 60° C. and bottled. The product is orange-red, clear and partially solid at room temperature. Store in the dark at a temperature of 25 ° C.

A value of 11.9 FNU was obtained as an average value of three measurements (11.4 FNU; 10.5　FNU; 13.9 FNU) by measuring the turbidity under physiological conditions (pH 1.1; 37° C.) after diluting with water at a ratio of 1:50.

Particle size of a sample of 20 microliters at a dilution ratio of 1:50 in a solution of sodium azide (200 mg/L) and saline (50 mmol/L) by Wyatt Technology Europe GmbH in the Eclipse program, i.e. by field flow fractionation. As a result of the analysis, a peak with a radius of 16.5 nm was obtained for the micelle of this solubilizate. Therefore, the diameter of the micelle is 33 nm.

10% α-liponic acid solubilizate

The following were used:

896.7 g Polysorbate 80,

103.3 g α-lipoic acid.

Heat the polysorbate 80 to 28-32°C. Under stirring, α-lipoic acid powder is added to the polysorbate and incorporated. The powder is added at such a rate that the powder is drawn into the emulsifier homogeneously during stirring. It is then heated to a temperature in the range of 83°C to 87°C. Once a homogeneous and transparent product is obtained, it is cooled to a temperature below 60° C. and bottled. The product is yellow and viscous and stored in the dark at a temperature of 25°C.

The product can also be prepared using polysorbate 20 or a mixture of polysorbate 80 and polysorbate 20.

At a pH of 1.1 and diluted 1:50 with water at a temperature of 37°C, this solubilize exhibited an average turbidity of 2.9 FNU.

Particle size of a sample of 20 microliters at a dilution ratio of 1:50 in a solution of sodium azide (200 mg/L) and saline (50 mmol/L) by Wyatt Technology Europe GmbH in the Eclipse program, i.e. by field flow fractionation. As a result of the analysis, a peak with a radius of less than 10 nm was obtained for the micelle of this solubilizate. Therefore, the diameter of the micelle is 20 nm.

10% xanthohumol-flavonoid pure solubilizate in ethanol (corresponding to 9.2% xanthohumol)

The following was used for this modification of the xanthohumol solubilizate according to the present invention:

100 g Xantho-Flav Pure (= 92 g xanthohumol),

150 g ethanol (96%) of neutral alcohol grade 1411 U, and

750 g Polysorbate 80.

First, Xantho-Flav Pure powder is dissolved in ethanol while heating to a temperature ranging from 48 to 52°C. A homogeneous solution is produced. Polysorbate 80 is then added to the solution of Xantho-Flav Pure in ethanol while heating to 83-87°C. The addition is carried out at such a rate that both fluids are homogenized under agitation. The obtained solubilized product is cooled and bottled at less than 60°C, and stored in a dark room at 25°C or less.

15% glavonoid solubilization with glycerol (= 0.45% glabridin)

The following were used:

150 g glavonoids (= 4.5 g glabridin),

100 g 99% glycerol,

750 g Polysorbate 80.

First, glycerol and glavonoids were mixed and homogenized at a temperature ranging from 18 to 22 °C. While heating and stirring to 83-87° C., polysorbate 80 was added to the fluid consisting of glycerol and glavonoids. Stir vigorously enough to obtain a homogeneous solubilisate, cool and bottle up to 30°C, then store in the dark below 25°C.

The aforementioned solubilisates can be used to prepare a solubilisate according to the invention comprising curcumin and at least one further active substance by mixing. This is described below with reference to exemplary embodiments 3, 5, 6, and 7.

Exemplary Embodiment 1

5.4% curcumin / 6.6% boswellic acid soluble cargo

This exemplary embodiment of a solubilizate according to the present invention was prepared directly. Active substances were co-micelleted. For this purpose the following was used:

82 g 80% Boswellia serrata extract (= 65.6 g Boswellic acid),

57 g 95% curcumin powder (= 54.1 g of curcumin),

70 g water,

350 g polysorbate 20,

441 g Polysorbate 80.

While heating to a temperature in the range of 48 to 52° C., polysorbate 20 and polysorbate 80 are mutually homogenized and mutually dissolved under stirring. While maintaining this temperature, the emulsifier mixture is mixed with the water while stirring vigorously enough so that the water and ethanol are uniformly dissolved in the emulsifier solution. At the same temperature, under stirring , the Boswellia serrata extract is incorporated into the emulsifier diluted with water. Boswellia serrata extract is added at a slow rate so that it is evenly aspirated into the dilute emulsifier solution under stirring. The temperature is then increased to the range of 63° C. to 67° C. under vigorous stirring. Curcumin powder is incorporated under agitation. The temperature is further increased to a value in the range of 85° C. to 89° C. while stirring vigorously enough to homogenize the curcumin evenly distributed in the formulation.

When diluted 1:500 in water at pH 1.1 and at a temperature of 37°C, the solubilize exhibits an average turbidity FNU of 1.9.

In the context of the present application, verification of whether the homogenization of the components for the formation of the solubilize according to the present invention is sufficiently complete in the preparation of any solubilisate is by measuring the clarity of the product, which indicates complete micellization. This is achieved using a laser beam. Such laser beam measurement involves, for example, illuminating the sample using a commercially available laser pointer, in particular using a wavelength ranging from 650 nm to 1700 nm (spectral color red), followed by visual inspection of the illuminated or irradiated soluble cargo. can be performed by Verification is accomplished within the reaction vessel rather than sampling and thus outside the reaction vessel. The laser beam is projected through a viewing window positioned in front of the reaction vessel and perpendicular to the reaction vessel. If only light spots appear on the inner surface of the rear surface of the reaction vessel and there is no scattering, it is a visual confirmation that the micellization process has been completed, since the resulting particle structure in the reaction vessel is smaller than the wavelength of visible light.

In the context of the present invention, the content of curcumin and boswellia extract in individual solubilates can also be adjusted significantly higher than the example shown, depending on the application.

Exemplary Embodiment 2

Soluble cargo of 3.3% curcumin / 3.6% boswellic acid and 1.8% xanthohumol

The following were used:

45 g 80% Boswellia serrata extract (36 g boswellic acid),

35 g 95% curcumin powder (33.25% curcumin),

23 g Xantho-Flav with at least 80% xanthohumol (18.4 g xanthohumol),

60 g water,

50 g ethanol (96%) neutral alcohol, grade 1411U,

350 g polysorbate 20,

437 g Polysorbate 80.

While heating to a temperature in the range of 48 to 52° C., polysorbate 20 and polysorbate 80 are mutually dissolved under stirring while homogenizing each other. While maintaining this temperature, the emulsifier mixture is mixed with water and ethanol. The emulsifier mixture is stirred vigorously enough so that the water and ethanol are uniformly dissolved in the emulsifier solution. At the same temperature, under stirring, the Boswellia serrata extract and xanthohumol are incorporated into the emulsifier mixture diluted with water while stirring. This addition is carried out at a sufficiently slow rate so that the Boswellia serrata extract and xanthohumol are uniformly drawn into the dilute emulsifier solution under agitation. The temperature is then increased to the range of 63° C. to 67° C. under vigorous stirring. Curcumin powder is incorporated under agitation. The temperature is further increased to a value in the range of 85° C. to 89° C. while stirring vigorously enough to homogenize the curcumin evenly distributed in the formulation.

Next, it is cooled to a temperature of 45° C. or lower. Then, a dark yellow viscous preparation containing curcumin and solubilized products of boswellic acid and xanthohumol is bottled and stored in a cool dark room below 25°C.

At a dilution ratio of 1:500 in water at pH 1.1 and a temperature of 37°C, the solubilize exhibits an average turbidity FNU of 1.9.

For particle size analysis of the solubilisate according to the present invention, unless otherwise stated, the solubilisate was first diluted with distilled water in a ratio of 1:500 and brought to 37° C. using a hotplate with constant stirring with a magnetic stirrer. . The pH was then adjusted to 1.1 using 32% hydrochloric acid. Samples were then measured immediately. The results are summarized in the table below.

Exemplary Embodiment 3

1.5% curcumin/3% boswellic acid/2% xanthohumol/0.35% serrapeptase solubilizate

According to the formulation described above, the following were used:

250 g 7% Curcumin Soluble,

250 g 12% boswellia solubles,

250 g 9.2% xanthohumol solubles, and

250 g 1.5% serrapeptase solubilizate.

By heating all four solubilizers at a temperature in the range of 50 ° C to 60 ° C, the viscosity can be lowered and the fluidity can be improved. Then, they are mixed together by stirring. As soon as a homogeneous complete product is obtained, it is optionally cooled to a temperature below 60° C. and bottled.

Prior to further processing, such as capsule filling, the product is preferably stirred again to homogenize and, if necessary, moderately heated, i.e. brought to a temperature of about 40° C. to 50° C.

The solubilize exhibits an average turbidity of 1.0 FNU when diluted 1:500 with water at a temperature of pH 1.1 and a temperature of 37°C.

The particle size analysis results are summarized in the following table.

*

Exemplary Embodiment 4

Soluble product of 5% glavonoids (=1.8% glabridin)/ 3% curcumin/ 3.5% xanthohumol

The following were used:

32 g 95% curcumin powder (= 30.4 g of curcumin),

44 g Xantho-Flav Pure powder (= 35.2 g xanthohumol),

60 g Kaneka glavonoids (= 1.8 g glabridin),

60 g 96% ethanol, neutral alcohol grade 1411U,

44 g 99.5% glycerol,

760 g Polysorbate 80.

Polysorbate 80 and glycerol are mixed while heating and stirring to a temperature ranging from 48 to 52° C. and the mixture is suitably homogenized. Ethanol is incorporated into the polysorbate-glycerol mixture while holding the temperature constant and stirring vigorously enough to form a homogeneous solution. The temperature is then raised to 63-67° C. while the xanthohumol is incorporated into the solution of polysorbate, glycerol, and ethanol while stirring vigorously enough to homogeneously combine the xanthohumol with the solution prepared above.

The curcumin powder is then incorporated into the xanthohumol solubilisate while raising the temperature to the range of 78-82°C. As with the incorporation of the glavonoids and the incorporation of the xanthohumol described below, stir vigorously enough so that the newly added solubilize components are homogeneously combined with the solubilized product in the prepared fluid. For the addition of glavonoids, the temperature is further raised to a temperature range of 85 to 98°C.

The product is a solubilization of co-micelleted curcumin, xanthohumol, and glavonoids. It was cooled to a maximum of 45° C. while stirring and then bottled.

Exemplary Embodiment 5

1.5% curcumin / 2% xanthohumol / 3.5% glavonoids (0.1% glabridin ) / 1.2% coenzyme Q ₁₀ solubles

Depending on the formulation, the following were used:

250 g 7% Curcumin Soluble,

250 g 9.2% xanthohumol solubles,

250 g 15% glavonoid solubilisate, and

250 g 5% Coenzyme Q10 Soluble Cargo.

All four solubilisates can be heated to a temperature range of 50° C. to 60° C. to lower the viscosity and thus improve fluidity. Then, they are mixed together by stirring. Once a homogeneous complete product is obtained, it is optionally cooled below 60° C. and bottled.

Prior to further processing, such as filling into capsules, it is preferred to stir it once again to homogenize it and, if necessary, heat it to a temperature suitable for this purpose, i.e. from about 40° C. to 50° C.

The particle size analysis results are summarized in the following table.

Exemplary Embodiment 6

1.3% curcumin / 1.6% xanthohumol / 3% glavonoids (0.09% glabridin ) / 1% coenzyme Q ₁₀ / 2% α-liponic acid solubles

Depending on the formulation, the following were used:

200 g 7% curcumin solubilize,

200 g 9.2% xanthohumol solubles,

200 g 15% glavonoid solubilisate,

200 g 5% coenzyme Q ₁₀ solubles, and

200 g 10% α-lipoic acid solubilize.

All five solubilisates can be heated to a temperature range of 50° C. to 60° C. to lower the viscosity and thus improve fluidity. Then, they are mixed together by stirring. Once a homogeneous complete product is obtained, it is optionally cooled below 60° C. and bottled.

Prior to further processing, such as filling into capsules, it is preferred to stir it once again to homogenize it and, if necessary, heat it to a temperature suitable for this purpose, i.e. from about 40° C. to about 50° C.

The particle size analysis results are summarized in the following table.

Exemplary Embodiment 7

Soluble in 0.8% Curcumin / 1.5% Boswellic Acid / 1% Xanthohumol / 1.8% Glavonoids (0.05% Glabridin) / 0.18% Serrapeptase / 1.2% Resveratrol / 0.6% Coenzyme Q10 / 1.2% α-Liponic Acid freight

Depending on the formulation, the following were used:

125 g 7% Curcumin Soluble,

125 g 12% boswellia solubles,

125 g 9.2% xanthohumol solubles,

125 g 15% glavonoid solubilisate,

125 g 1.5% serrapeptase solubilizate,

125 g 10% resveratrol solubles,

125 g 5% coenzyme Q ₁₀ solubles, and

125 g 10% α-lipoic acid solubilize.

All eight solubilisates can be heated to a temperature range of 50° C. to 60° C. to lower the viscosity and thus improve fluidity. Then, they are mixed together by stirring. Once a homogeneous complete product is obtained, it is optionally cooled below 60° C. and bottled.

Prior to further processing, such as filling into capsules, it is preferred to stir it once again to homogenize it and, if necessary, heat it to a temperature suitable for this purpose, i.e. from about 40° C. to 50° C.

Upon dilution with water at a pH of 1.1 and a dilution ratio of 1:50 at a temperature of 37 °C, the solubilisate exhibits an average turbidity of 12.5 FNU.

The particle size analysis results are summarized in the following table.

Exemplary Embodiment 8

0.8% Curcumin / 1.5% Boswellic Acid / 1% Xanthohumol / 1.8% Glavonoids (0.05% Glabridin) / 0.18% Serrapeptase / 1.2% Resveratrol / 0.6% Coenzyme Q ₁₀ / 1.2% α-Liponic Acid Direct Manufacturing Available Cargo

The following was used:

9.375 g 95% curcumin powder,

19 g Boswellia Serrata extract (15.2 boswellic acid),

12.5 g Xantho-Flav powder (at least 80% xanthohumol = 10 g xanthohumol),

18.75 glavonoids (0.56 g glabridin),

1.875 g Serrapeptase (37,500,000 U),

12.5 g resveratrol,

7.19 g coenzyme Q ₁₀ ,

12.5 g α-lipoic acid,

7.875 g water,

18.75 g ethanol,

12.5 g glycerol,

9.375 g mixed tocopherols,

27.685 g MCT oil,

122.5 g polysorbate 20, and

707.225 g Polysorbate 80.

Polysorbate 80 is mixed with α-lipoic acid at a temperature in the range of 18 to 22° C. with sufficiently vigorous stirring to produce a homogeneous mixture. Separately, serrapeptase is dissolved in water in the same manner at a temperature range of 18 to 22°C. Ethanol, xanthohumol, curcumin, boswellia, glavonoids, resveratrol, coenzyme Q ₁₀ , mixed tocopherols, glycerol, MCT oil, and polysorbate as additional ingredients while still maintaining the temperature range at 18-22° C. 20 is added slowly and continuously to the mixture of serrapeptase and water. Care is taken to ensure that the product is well and homogeneously mixed. If appropriate, add one ingredient and rest before adding another ingredient. The addition and stirring are carried out slowly enough so that the specific component to be added is homogeneously incorporated into the formulation.

The two mixtures, namely the formulation of polysorbate 80 and α-liponic acid and the mixture of the other ingredients, are then mixed together and further stirred at a temperature in the range of 18 to 22°C. This gives a homogeneous pasty mass resembling a slurry, which is heated to a temperature range of 85°C to 89°C. Heating is carried out with constant and sufficiently slow stirring so that the heated slurry remains as homogeneous as possible all the time. After cooling to a temperature below 60° C., the product is bottled. It is dark and viscous and stored in the dark at temperatures below 25°C. Prior to further processing, such as filling into capsules, it is preferred to stir it once again to homogenize it and, if necessary, heat it to a temperature suitable for this purpose, i.e. from about 40° C. to about 50° C.

Upon dilution with water at a pH of 1.1 and a dilution ratio of 1:50 at a temperature of 37 °C, the solubles exhibit an average turbidity of 1.0 FNU.

The particle size analysis results are summarized in the following table.

Exemplary Embodiment 9

3% Curcumin / 3.2% Boswellic Acid / 1.6% Xanthohumol / 1% Solubilizes of CBD Oil

Use:

40.5 g 80% Boswellia serrata extract (32.4% Boswellic acid),

31.5 g 95% curcumin powder (29.925 g curcumin),

20.7 g Xantho-Flav powder containing at least 80% xanthohumol (16.5 g xanthohumol),

54 g water,

45 g ethanol,

315 g polysorbate 20,

483 g Polysorbate 80,

10 g CBD oil: 30% cannabidiol

Cannabidiol (CBD) is a barely psychoactive cannabinoid derived from the female cannabis Cannabis sativa or Cannabis indica . Non-THC-free CBD oil was used, meaning traces of THC may be present. Tetrahydrocannabinol (THC) is responsible for the hallucinogenic effects of the cannabis plant.

While heating to a temperature in the range of 48 to 52° C., polysorbate 20 and polysorbate 80 are homogenized while dissolving each other by stirring. While maintaining the temperature, the emulsifier mixture is mixed with water and ethanol. Stir vigorously enough so that the water and ethanol are evenly dissolved in the emulsifier solution. At the same temperature, the Boswellia serrata extract and xanthohumol are incorporated into the diluted emulsifier mixture with water while stirring. The addition is carried out at a rate slow enough so that the Boswellia serrata extract and xanthohumol are drawn up uniformly into the diluted emulsifier solution under agitation. The temperature is then raised to a range of 63° C. to 67° C. under vigorous stirring. Curcumin powder is incorporated while stirring. The temperature is further raised to a value in the range of 85° C. to 89° C. while stirring vigorously enough to homogenize and distribute the curcumin uniformly in the formulation. The CBD oil is then incorporated into the mixture while stirring vigorously enough so that the CBD oil is evenly distributed in the formulation and homogenized.

It is then cooled to a temperature below 45°C. A dark yellow viscous formulation comprising solubilizers of curcumin and boswellic acids and xanthohumol and CBD oil is bottled and stored in the dark and cold, i.e. at a temperature below 25°C. The solubilizate and its aqueous solution are stably homogeneous and in crystal clear soluble form, like the solubilisate according to illustrative embodiment 2.

It will be apparent to those skilled in the art that the present invention is not limited to the embodiments described above and can be modified in many ways. Combining or exchanging features of the individually exemplified embodiments is particularly possible.

Claims (25)

As a soluble cargo, the soluble cargo is
curcumin in an amount of 3% to 7% by weight;
Xanthohumol, boswellia, glavonoids, glabridin, cannabinoids, serrapeptase, coenzyme Q ₁₀ , α-lipoic acid, up to 10% by weight, including one or more substances selected from the group consisting of resveratrol amount of at least one additional active substance; and
polysorbate 80 or polysorbate 20 or a mixture of polysorbate 20 and polysorbate 80, wherein the polysorbate content ranges from 75% to 95% by weight;
ethanol in an amount of 0 to 20% by weight;
glycerol in an amount of 0 to 22% by weight; and
Water in an amount of 0 to 10% by weight
contains,
treatment and/or prevention of inflammation, cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, elevated blood sugar, diabetes, metabolic syndrome, and/or autoimmune disease, multiple sclerosis (MS), visceral fat reduction, thermogenesis, Cholesterol reduction, LDL cholesterol reduction, and/or blood glucose (blood sugar) reduction and/or blood triglyceride reduction, macular pigment density improvement, oxidative stress reduction and/or fat accumulation in hepatocytes, fatty liver disease, Friedreich's ataxia, lysosomal A soluble cargo intended for use as a pharmaceutical for the treatment and/or prevention of disease, Tay-Sachs disease, arteriosclerosis, heart disease, arthritis. According to claim 1,
As an anti-inflammatory drug and/or as a drug effective for cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, increased blood sugar, diabetes, metabolic syndrome, and/or autoimmune disease, multiple sclerosis (MS), reducing visceral fat , as a medicine for reducing fever, cholesterol reduction, LDL cholesterol, and/or blood glucose (blood sugar) reducing effect and/or blood triglyceride reducing effect, macular pigment density improving effect, oxidative stress reducing effect and/or fat in hepatocytes A soluble cargo intended for use as a pharmaceutical which is effective for accumulation reduction, fatty liver disease, Friedreich's ataxia, lysosomal disease, Tay-Sachs disease, arteriosclerosis, heart disease, and arthritis. 2. The solubilize according to claim 1, wherein the solubilizate contains up to 15% by weight of ethanol. 2. The solubilizate according to claim 1, wherein the solubilizate contains at most 10% by weight of glycerol. 2. The solubilizate according to claim 1, wherein the solubilizate additionally contains up to 7% by weight of water. The method according to claim 1, wherein the diameter distribution of the micelles in the diluent of the solubilizate diluted with distilled water at a ratio of 1:500 under physiological conditions (pH 1.1 and 37 ° C) is between d ₁₀ = 6 nm and d ₉₀ = 20 nm. available cargo. 2. The method of claim 1, wherein the turbidity of the solubilisate is measured by infrared radiation according to the ISO 7027 standard specification for a dilution of the solubilisate diluted with water in a ratio of 1:50 or 1:500 under physiological conditions (pH 1.1 and 37°C). A soluble cargo that is less than 25 FNU, or less than 3 FNU, as measured by light scattering measurements using A capsule filled with the solubilizate according to any one of claims 1 to 7, wherein the capsule is a soft gelatin capsule or a hard gelatin capsule or a soft gelatin-free capsule or a hard gelatin-free capsule or a cellulose capsule. A form of capsule. A fluid containing the solubilizate according to any one of claims 1 to 7, wherein the fluid is selected from the group consisting of foods, dietary supplements, beverages, cosmetics and pharmaceuticals. 10. The fluid of claim 9, wherein the fluid comprises an aqueous dilution of the solubilizate. treatment and/or prevention of inflammation, cancer, Alzheimer's disease, Parkinson's disease, obesity, hypercholesterolemia, elevated blood sugar, diabetes, metabolic syndrome, and/or autoimmune disease, multiple sclerosis (MS), visceral fat reduction, thermogenesis, Cholesterol reduction, LDL cholesterol reduction, and/or blood glucose (blood sugar) reduction and/or blood triglyceride reduction, macular pigment density improvement, oxidative stress reduction and/or fat accumulation in hepatocytes, fatty liver disease, Friedreich's ataxia, lysosomal The soluble cargo according to any one of claims 1 to 7, which is used in a method for the treatment and/or prevention of disease, Tay-Sachs disease, arteriosclerosis, heart disease, arthritis, As a composition comprising,
The method is to orally administer a composition comprising the solubilizate according to any one of claims 1 to 7 to a dietary supplement consumer or patient. 12. The composition of claim 11, wherein the solubilizate is administered to the dietary supplement consumer or patient at a dose ranging from 0.5 mg/kg body weight to 1 mg/kg body weight of curcumin, or at a dose of 0.81 mg/kg body weight once daily. 12. The composition of claim 11, wherein the solubilizate is administered to the dietary supplement consumer or patient at a dose of Boswellia 1 mg/kg body weight to 2 mg/kg body weight, or at a dose of 1.62 mg/kg body weight once daily. 12. The composition of claim 11, wherein the solubilizate is administered to the dietary supplement consumer or patient at a dose of 0.5 mg/kg body weight to 1 mg/kg body weight of xanthohumol, or at a dose of 0.81 mg/kg body weight. A method for producing the solubilizate according to any one of claims 1 to 7, the method comprising:
(a) providing polysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20 and polysorbate 80;
(b) adding at least one additional active substance selected from the group consisting of xanthohumol, boswellia, glavonoids, cannabinoids, serrapeptase, coenzyme Q ₁₀ , α-lipoic acid, and resveratrol;
(c) adding curcumin powder;
wherein step (a) comprises heating the temperature to a temperature range of 40°C to 62°C, or a temperature range of 45°C to 57°C, or a temperature range of 48°C to 52°C; and
Step (b) involves holding the temperature unchanged compared to step a), or heating to a temperature range of 60°C to 75°C, or a temperature range of 61°C to 70°C, or a temperature range of 63°C to 67°C. contains; and
Wherein step (c) comprises heating to a temperature range of 82°C to 97°C, or a temperature range of 83°C to 92°C, or a temperature range of 85°C to 89°C. 16. The method according to claim 15, wherein the additional active substance in step (b) is Boswellia and/or Xanthohumol, and Boswellia serrata extract is used as Boswellia. 16. The method of claim 15, prior to step (b), adding water in a temperature range of 40 °C to 62 °C, or in a temperature range of 45 °C to 57 °C, or in a temperature range of 48 °C to 52 °C (b1) How to do it. 18. The method of claim 17, wherein step (b1) further comprises adding ethanol at a temperature ranging from 40 °C to 62 °C, or at a temperature ranging from 45 °C to 57 °C, or at a temperature ranging from 48 °C to 52 °C. A method for producing the solubilizate according to any one of claims 1 to 7,
(a) preparing a first formulation by providing polysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20 and polysorbate 80 and α-lipoic acid as at least one first active substance; doing;
(b) preparing a second formulation by providing water and serrapeptase as at least one additional active substance;
(c) adding at least one further active substance selected from the group consisting of xanthohumol and/or Boswellia serrata extract and/or glavonoids and/or resveratrol and/or coenzyme Q ₁₀ , adding curcumin powder to the second formulation from step (b);
(d) combining the first formulation from step (a) and the second formulation from step (c), wherein the temperature during steps (a) to (d) is in the range of 18° C. to 22° C. phosphorus step;
(e) heating to a temperature range of 80° C. to 97° C., or a temperature range of 83° C. to 92° C., or a temperature range of 85° C. to 89° C.
How to include. 20. The method of claim 19, wherein step (c) is ethanol and/or glycerol and/or MCT oil and/or polysorbate 20 and/or polysorbate 80 and/or a mixture of polysorbate 20 and polysorbate 80 A method further comprising adding a. A method for producing the solubilizate according to any one of claims 1 to 7,
(a) providing a mixture of polysorbate 80 and/or polysorbate 20 and/or polysorbate 20 and polysorbate 80 and a mixture of glycerol and ethanol;
(b) adding an ethanolic extract of hard resin from hops as a source of xanthohumol as at least one further active substance,
step (a) includes heating to a temperature range of 40°C to 62°C, or a temperature range of 45°C to 57°C, or a temperature range of 48°C to 52°C;
Step (b) comprising heating to a temperature range of 60 ° C to 75 ° C, or a temperature range of 61 ° C to 70 ° C, or a temperature range of 63 ° C to 67 ° C; and
(c) adding curcumin powder at a temperature range of 70°C to 92°C, or a temperature range of 75°C to 87°C, or a temperature range of 78°C to 82°C;
(d) adding glavonoids while heating to a temperature range of 80° C. to 97° C., or a temperature range of 83° C. to 92° C., or a temperature range of 85° C. to 89° C.
How to include. A method for preparing the solubilizate according to any one of claims 1 to 7, wherein curcumin solubilisate and a solubilisate of at least one additional active substance are mixed, and these individual solubilisates are mixed in a weight ratio of 1:1 The method of mixing with . A method for producing a xanthohumol solubilisate for use in the method according to claim 22,
(a) providing an ethanolic extract of hard resin from hops, or Xantho-Flav Pure powder, as a source of xanthohumol;
(b) adding polysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20 and polysorbate 80;
wherein step (b) comprises heating to a temperature range of 80°C to 95°C, or a temperature range of 81°C to 90°C, or a temperature range of 83°C to 87°C. The method of claim 23, prior to step (b), the following (b1) step, namely
(b1) Ethanol extract of hard resin from hops, or Xantho-Flav Pure powder, as a source of xanthohumol, at a temperature range of 40 ° C to 62 ° C, or a temperature range of 45 ° C to 57 ° C, or 48 ° C to 52 ° C A method comprising dissolving in ethanol while heating to a temperature range of A method for preparing a serrapeptase solubilizate for use in the method of claim 22, the method comprising:
(a) providing water and serrapeptase;
(b) adding MCT oil;
(c) adding polysorbate 80 and/or polysorbate 20 and/or a mixture of polysorbate 20 and polysorbate 80
Including,
The temperature during step (a) ranges from 18° C. to 22° C.;
Wherein step (b) comprises heating to a temperature range of 77°C to 93°C, or a temperature range of 80°C to 90°C, or a temperature range of 83°C to 87°C.