CN101453914A - Lycopene for the treatment of metabolic dysfunction - Google Patents

Abstract

This invention relates to the treatment of metabolic dysfunction and disorders associated with metabolic dysfunction using lycopene compounds. Methods of treatment and uses of lycopene compounds in such methods are provided.

Description

The lycopene of treatment metabolic function disorder

The present invention relates to treat the method and the material of the metabolic function disorder that comprises insulin resistance, glucose tolerance, hypertension, PCOS, obesity, steatosis, chronic hepatitis and cirrhosis.

Unusually become more and more general in developed country such as metabolism such as insulin resistance, GI, hypertension, obesity and metabolic syndromes, according to estimates, only just have 50,000,000 people of surpassing to suffer from this class dysfunction in the U.S..The metabolic function disorder is secondary diabetes, angiocardiopathy, the important risk factor of obliteran, brain and other forms of arteriosclerosis on every side.

The further exploitation of effective methods of treatment of these states had significant impact to the whole mankind's health.

The inventor finds lycopene external, and disorder has positive effect to metabolic function to animal model with in human clinical trials.

One aspect of the present invention provides the method for treatment metabolic function disorder, and it comprises with the lycopene compound for the treatment of effective dose its individual administration of needs.

The lycopene compound can comprise lycopene and the lycopene derivative with similar lycopene biological property.Lycopene is the unsaturated C of the open chain of structure I ₄₀Carotenoid (chemical abstracts service registration number be 502-65-8), it results from natively such as tomato, guava, rose is real, the plant of watermelon and pink grape fruit.

The lycopene that is used for purposes described herein can comprise one or more different isomers.For example, lycopene can comprise at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, (Z)-isomers of at least 90% or at least 95%, (full E)-isomers, the cis-isomer that perhaps has the bioavilability of improvement with respect to transisomer is such as 5-cis-isomers, or 9-cis-isomers, or 13-cis-isomers.Transisomer can be in vivo or store and process in isomery turn to cis-isomer.

The lycopene derivative that has with the similar biological property of lycopene can comprise, for example, such as vitamin A acid, synthetic acyclic vitamin A acid or 1-HO-3 ', 4 '-two dehydrogenation lycopenes, 3,1 '-(HO) 2-gamma carotene, 1,1 '-(HO) 2-3,4,3 ', 4 '-four dehydrogenation lycopenes and 1,1 '-(HO) 2-3, the carotenoid of the two dehydrogenation lycopenes of 4-.

The lycopene compound that is used for purposes described herein can be natural, promptly takes from natural origin, for example, extracts from the plant such as tomato or melon.Extraction from plant, serial of methods concentrated and/or purifying lycopene compound are well known in the art.For example, can use the solvent extraction method that uses ethanol, dimethyl sulfoxide (DMSO), ethyl acetate, hexane, acetone, soybean or other vegetable oil or non-plant oil.

The lycopene compound that is used for purposes described herein can synthesize, promptly by artificial means preparation, for example, by chemical synthesis.A series of chemical synthesis process of lycopene and other carotenoid are well known in the art.For example, the synthetic carotenoid of available three grades of chemical synthesis based on standard Wittig olefination scheme wherein generates the C that is dissolved in carrene (DCM) ₁₅Phosphine mesylate organic solution and the C that is dissolved in toluene ₁₀Dialdehyde organic solution, these two kinds of solution combine with sodium methoxide solution step by step and generate rough lycopene by condensation reaction.Can use the lycopene of routine techniques purification of crude then, for example add glacial acetic acid and deionized water in mixture, vigorous stirring makes water separate with organic facies, and the water extraction contains the organic facies of DCM and rough lycopene.Methyl alcohol is joined in the organic facies, and remove DCM by decompression distillation.With crude methanol lycopene solution heating and be cooled to magma, this magma is filtered and uses methanol wash subsequently.Can be recrystallized subsequently and with the dry lycopene crystal of hot nitrogen.Synthetic lycopene also can be bought from businessman (for example, BASF Corp, NJ USA).

With respect to natural lycopene, synthetic lycopene can contain the more cis-isomer of vast scale.For example, synthetic lycopene can contain 5-cis-isomer up to 25%, 1% 9-cis-isomer, 1% 13-along isomers, other cis-isomers of 3%, and can contain the 9-cis-isomer, 1% 13-cis-isomer of 5-cis-isomer, the 0%-1% of 3%-5% and less than other cis-isomers of 1% from the lycopene that tomato produces.Since All-cislycopene has higher bioavilability with respect to trans lycopene, so for some purpose, synthetic lycopene is preferred.

The derivative of above-mentioned lycopene can be by being similar to above-mentioned synthetic chemical synthesis or producing by the natural lycopene that extracts from vegetable material is carried out chemical modification.

The lycopene compound can be by any form easily or preparation administration.Appropriate formulations can promote or strengthen the absorption and the interior bioavilability of body thereof of lycopene compound.For example, the lycopene compound can be used as the pharmaceutical composition administration, it comprises lycopene, with one or more pharmaceutically acceptable carriers, adjuvant, excipient, diluent, filler, buffer solution, stabilizing agent, emulsifying agent, anticorrisive agent, lubricant or well known to a person skilled in the art other materials, and randomly and other treatment or preventative preparation.For example, in some embodiments, the lycopene compound can be used as the pharmaceutical composition administration, and it comprises lycopene compound and isoflavones, for example, and isoflavones and/or vitamin C.

As described herein, the appropriate drug composition can comprise the lycopene compound as the single-activity component, also can be by the lycopene compound is mixed to come preparation with one or more pharmaceutically acceptable carriers, excipient, buffer solution, adjuvant, stabilizing agent or other materials.

Term used herein " pharmaceutically acceptable " relates to compound, material, composition and/or formulation, in rational medical assessment scope, its be suitable for individual (for example, the mankind) tissue contacts, and not having too much toxicity, excitant, allergic reaction or other problems or complication, it matches with rational benefit/risk ratio.On with the compatible meaning of other preparation compositions, every kind of carrier, excipient etc. also all must be " acceptable ".

Suitable carrier, excipient etc. can find in the standard pharmaceutical textbook, for example, and Remington ' s Pharmaceutical Sciences (Lei Shi pharmacy complete works), 18 ^ThEdition, MackPublishing Company, Easton, Pa., 1990, and can comprise oils, and for example, such as the vegetable oil of tomato oil, soybean oil or peanut oil, perhaps non-plant oil, glycerine, gelatin, sucrose, glucose, ascorbyl palmitate, cornstarch and silica.Suitable emulsifying agent comprises polysorbate 80.Suitable stabilizers can comprise sucrose ester, lecithin, and such as the antioxidant of dl-alpha-tocopherol and ascorbyl palmitate, flavones, cellulose, wax, sweet mellow wine, shellac, mica, calcium phosphate, dolomol and Arabic gum or acacia gum.

For example, lycopene can be formulated to pearl, tablet or other solids, it comprises about 10% lycopene, be respectively about 1.5% and 5.0% antioxidant dl-a-tocopherol and ascorbyl palmitate, also has the carrier mass such as vegetable oil, isinglass, sucrose and cornstarch in addition.

The appropriate formulation of lycopene can and comprise LycoVit from businessman's purchase ^TM10, LycoVit ^TM10 cold water dispersant (CWD) and LycoVit ^TMDispersant 20 percent (all from BASF Corp, NJ, USA), Lyc-O-Mato ^TM(LM), LycoBeads ^TMAnd Tomato-O-Red ^TM(Dalidar Pharma, LycoRed Ltd UK).

In some embodiments, can be with lycopene compound and solubilizer preparation together.Solubilizer comprises hydrophilic compounds, and it dissolves in water-based solution, also can be insoluble to organic solvent.Suitable hydrophilic solvents comprises soluble protein, particularly such as casein, beta lactoglobulin, ALA and sero-abluminous lactoprotein.Lactalbumin can be used as solubilizer easily.Lactalbumin is isolated many globular preteinses from whey, and they are the byproducts when milk is made cheese.Lactalbumin is the mixture of beta lactoglobulin (～65%), ALA (～25%) and seralbumin (～8%), and their native form does not rely on pH and be solvable.With the lycopene formulations (being called " newborn lycopene (lactolycopene) ") of lactalbumin is well known in the artly (to see, for example, Richelle et al J.Nutr.132:404-408,2002 andEP-B-1289383 (PCT/EP01/06145)), and can buy (INNEOV from businessman, L ' Or é al (UK) Ltd, London).Preparation can comprise with the method for the lycopene formulations of lactalbumin, is being enough to form under the condition of mixture with lactalbumin and lycopene mixing.For example, lactalbumin is water-soluble, lycopene is dissolved in solvent such as acetone, ethanol or isopropyl alcohol, merge two solution, evaporate described solvent to breast-lycopene composition and form.

Selectively, can form composition like this: with lycopene and solvent to form first mixture, this first mixture is mixed to form second mixture with the lactalbumin of powdery or aqueous solution form, and the solvent that evaporates second mixture is to produce the composition as dispersant.The solvent that is fit to comprises acetone, ethanol and isopropyl alcohol.During with water and solvent, can use the solvent of 60:40 order easily: water volume ratio.Composition can comprise at the most 50% lycopene compound and 90% lactalbumin at the most.

This dispersant can be randomly through heat treatment to produce gel or spray-dried or freeze drying to produce powder.

As mentioned above, said composition subsequently can be together with carrier, excipient, stabilizing agent and emulsifying agent preparation.For example, Shi Yi newborn lycopene formulations can comprise the soybean lecithin of polysorbate80 and 1.5% (w/w) of silica, 3% (w/w) of microcrystalline cellulose, 5% (w/w) that percentage by weight (w/w) is the lactalbumin, 50.5% (w/w) of 2% lycopene, 20% (w/w).

The newborn lycopene formulations that is used for purposes described herein can obtain maybe can obtain as described above.

Can comprise obesity, insulin resistance, glucose tolerance reduction, PCOS (PCOS), hypertension with the metabolic function disorder of lycopene as herein described treatment, such as the fibrillatable of steatosis, chronic hepatitis, liver and the liver symptom of sclerosis, and metabolic syndrome.

In some embodiments, the individuality for the treatment of according to this method is not diabetes patient's (promptly not suffering from 1 type or diabetes B), and/or does not suffer from the atherosclerotic state such as CHD, apoplexy or peripheral vascular disease.

Obesity is that the body fat with surplus is the state of feature.For example, obese individuals can have the body mass index (BMI: body weight/height greater than 30 ²).Obesity is the risk factors of a series of medical conditions, and described state comprises diabetes and such as the cardiovascular symptom of atherosclerotic, ischemic (coronary artery) heart disease, myocardial ischemia (angina pectoris) and apoplexy.Obesity can comprise abdominal obesity, and it usually is defined as European man's waistline and surpasses 94 centimetres, and European woman's waistline is above 80 centimetres, South Asia man's waistline is above 90 centimetres, South Asia woman's waistline surpasses 80 centimetres, and Japanese man's waistline surpasses 85 centimetres, and skippy's waistline is above 90 centimetres.Methods described herein can be used for body weight control and bariatrician.

To be phalangeal cell, tissue or whole health can not show physiological reaction to the insulin of specified rate to insulin resistance.The result is with respect to contrast, needs more substantial insulin to reach the physiologic effect of insulin.Insulin resistance is present in 25% the non-diabetic people, and the estimation conversion rate from the insulin resistance state to diabetes B is annual 2%-12%.

If behind 120 minutes oral glucose tolerance test, the concentration of glucose in the blood plasma continues to be higher than 7.8mmol, the glucose tolerance has then taken place reduced.Glucose tolerance reduction is associated with insulin resistance and hyperglycemia.

PCOS (PCOS) is the endocrine symptom that influences 5%-10% women, and wherein the ovary vesica has disturbed the ovary circulation that normal hormone produces.The frequent same insulin resistance of PCOS, compensatory hyperinsulinemia, metabolic syndrome, obesity and high male sex hormone mass formed by blood stasis are associated.

It is feature that hypertension raises above 140mmHg and 90mmHg with the blood pressure continuation, the systolic pressure of (〉 140; 90 diastolic pressure).

The liver symptom comprises the fibrillatable and the sclerosis of steatosis, chronic hepatitis, liver.The liver fat sex change is being reduced and/or lipoprotein is synthetic and discharge the unusual lipid accumulation of liver that causes that descends be feature by fatty acid oxidation.But be converted into to the steatosis Secondary cases fibrillatable and the sclerosis of liver.Liver fibrosis is with the be grown to feature of fibr tissue liver.Fibrillatable can cause sclerosis, and wherein liver function can reduce and liver is permanently scabbed, the fibroid that becomes and greasiness.

Chronic hepatitis is meant the inflammation of liver, and it can be caused by the blood transfusion of bacterium or virus infections, parasitic infestation, alcohol, medicine, toxin or non-compatibility blood.

Metabolic syndrome is to be the state of feature with in high blood triglyceride levels and/or the low HDL cholesterol levels in obesity and insulin resistance, hypertension, the blood one or more.For example, IDF (International Diabetes Federation) is defined as metabolic syndrome: abdominal obesity is together with following any two: the triglyceride levels of rising (surpassing 1.7mmol/L), (man is in or is lower than 0.9mmol/L the HDL cholesterol that reduces, the woman is in or is lower than 1.1mmol/L), elevated blood pressure (being higher than 130/85) and the fasting plasma glucose (surpassing 5.6mmol/L) that raises.

Metabolic function disorder such as metabolic syndrome also can be associated with a series of states, and described state comprises hypercholesterolemia, low HDL cholesterol (for example, man,＜0.9mmol/l; The woman,＜1.0mmol/l), hypertriglyceridemia (for example, blood plasma TAG's, 〉=1.7mmol/l) and microalbuminuria (for example, urinary albumin excretion speed 〉=20 μ g/min; Globulin: the kreatinin ratio 〉=30mg/min).

Methods described herein can be used for the treatment with the disorderly associated state of metabolic function.For example, treatment can comprise with the method for the disorderly associated state of metabolic function: use the lycopene compound to its individual administration of needs.

Can comprise hypercholesterolemia, low HDL cholesterol, hypertriglyceridemia and microalbuminuria with the disorderly associated state of metabolic function.

Method of the present invention also can have prophylactic use.For example, prevent or postpone that the method that the metabolic function disorder shows effect can comprise that use lycopene compound is to its individual administration of needs.

Suitable experience is described herein to prevent or postpones that the individuality of the method that the metabolic function disorder shows effect can have the risk or the metabolic function disorder susceptible of suffering from the metabolic function disorder, for example, this individuality can have one or more with the relevant risk factors of the disorderly outbreak of metabolic function.

Methods described herein can comprise together with his spit of fland (statin) (being 3-hydroxy-3-methylglutaryl-coenzyme A (HMG Co A) reductase inhibitor) administration lycopene compound.

His spit of fland that is fit to comprises Pravastatin (PRAVACHOL ^TM), Lovastatin (MEVACOR ^TM), Simvastatin (ZOCOR ^TM), cerivastatin (LIPOBAY ^TM), Fluvastatin (LESCOL ^TM), Atorvastatin (LIPITOR ^TM), mevastatin and rosuvastatin (Crestor ^TM).Other his spits of fland that are fit to are known in this field and can use HMG-CoA reductase well known in the art analysis to identify like a cork.The example of these analyses has been disclosed in United States Patent (USP) the 4th, 231, No. 938.

It is disorderly or with the disorderly relevant state of metabolic function that another aspect of the present invention provides the lycopene compound to be used for the treatment of above-mentioned metabolic function in preparation, perhaps prevents or to postpone metabolic function disorderly or with the purposes in the medicine of the disorderly relevant state outbreak of metabolic function.

Another aspect of the present invention provides that to be used for the treatment of metabolic function disorderly or with the lycopene compound of the disorderly relevant state of metabolic function, and is used to prevent or to postpone metabolic function disorderly or with the lycopene compound of the disorderly relevant state outbreak of metabolic function.

Comprise lycopene compound compositions or medicine and can also comprise his spit of fland (being 3-hydroxy-3-methylglutaryl-coenzyme A (HMG Co A) reductase inhibitor).

His example in spit of fland that is fit to is addressed hereinbefore.Other his spit of fland that can be used for the inventive method is apparent to a skilled reader.

Another aspect of the present invention is a kit, and it comprises lycopene compound and the Ta Ting of coupling with the disorder of treatment metabolic function.

Kit can also comprise one or more other article and/or the reagent that are used to carry out the inventive method, described article and/or reagent such as with syringe with the lycopene compound separately or with the device (these components generally are aseptic) of other compound combination medicine-feeding.

This kit can also comprise the explanation of carrying out all or part of the inventive method, for example the administration system in lycopene compound and/or his spit of fland.

Yet another aspect of the present invention be lycopene compound and Ta Ting to be combined in the above-mentioned metabolic function of preparation treatment disorderly or with the disorderly relevant state of metabolic function, perhaps prevent or postpone purposes in the metabolic function medicine disorderly or that show effect with the disorderly relevant state of metabolic function.

The lycopene compound formulation that uses in this method can exist with unit dosage form easily, and can use the known any pharmaceutical methods preparation of pharmaceutical field.These class methods comprise the step of the carrier associating that makes reactive compound and constitute one or more auxiliary agents.Usually, the preparation of said preparation is by making reactive compound and liquid-carrier or the solid carrier that well separates or the two homogeneous and closely associating, then making product shaping if necessary.

The form of preparation can be liquid, solution, suspension, dispersant, emulsion, elixir, syrup, tablet, lozenge, granula, pulvis, capsule, cachet, pearl, pill, ampoule, suppository, pessary, ointment, gel, paste, creme, spray, mist agent (mists), foaming agent, lotion, finish, pill, electuary or aerosol.

(for example, by picked-up) preparation can exist with the discrete unit such as capsule, cachet or tablet suitable for oral administration, and each all comprises the reactive compound of scheduled volume; Described preparation can exist with pulvis or granula; Solution or suspension with water-based or non-aqueous liquid exist; Perhaps exist with oil-in-water liquid emulsion or Water-In-Oil liquid emulsion; Exist with pill; Exist with electuary; Perhaps exist with paste.Composition of liquid medicine generally comprises the liquid-carrier such as water, oil, animal or plant oil, mineral oil or artificial oil.Can comprise saline, glucose or other saccharide solution, perhaps such as the dihydroxylic alcohols of ethylene glycol, propane diols or polyethylene glycol.

Tablet can prepare by conventional means, for example, and randomly together with one or more auxiliary agent compressing tablet or moulding.Can be by in suitable machine, will preparing by the tablet of compressing tablet such as the reactive compound compressing tablet of the free-flowing form of pulvis or granula, described reactive compound randomly with one or more adhesives (for example, PVP, gelatin, acacia gum, sorbierite, tragacanth, hydroxypropyl methylcellulose), filler or diluent are (for example, lactose, microcrystalline cellulose, calcium monohydrogen phosphate), lubricant (for example, dolomol, mica, silica), disintegrant (for example, sodium starch glycollate, PVPP, Ac-Di-Sol), surfactant or dispersant or wetting agent are (for example, lauryl sodium sulfate) and anticorrisive agent (for example, right-methyl hydroxybenzoate, right-nipasol, sorbic acid) mixes mutually.Can in suitable machine, moulding prepare the tablet of moulding through the mixture of the wetting powder compound of inert liquid diluent.Tablet can be randomly by dressing or indentation, and can wherein use by preparation so that the reactive compound of slowly-releasing or controlled release to be provided, and for example, thereby the hydroxypropyl methylcellulose of various ratios provides required release type.Tablet can randomly provide with enteric coating, so as to be provided at intestines part but not in the release of the part of stomach.

For vein, skin or hypodermic injection, or in the injection at ailing position, active component can be with the form without the acceptable aqueous solution of intestines, it does not contain pyrogen and has suitable pH, isotonicity and stability.Utilize for example to wait and ooze media, this area person skilled can prepare suitable solution fully, and described grade is oozed media such as sodium chloride injection, ringer's injection, lactated Ringer's parenteral solution.As required, can comprise anticorrisive agent, stabilizing agent, buffer solution, antioxidant and/or other additives.

The example of above-mentioned technology and operation can be in Remington ' s PharmaceuticalSciences (Lei Shi pharmacy complete works), 16 ^ThEdition, Osol, A. (ed) finds in 1980.

Preferred use " treatment effective dose " administration, this is enough to show the benefit to individuality.The optimal dose that is appreciated that reactive compound and the composition that contains reactive compound can be different because of the patient.Determine that dose,optimum generally can relate to the level of therapeutic benefit of methods of treatment of the present invention and the balance of any risk or toxic and side effect.Selected dosage level can depend on various factors, include but not limited to: the activity of specific compound, method of administration, administration time, the compound rate of discharge, the treatment duration, the other drug of coupling, compound and/or material, and the previous medical history of age, sex, body weight, state, general health and patient.Can obtain Expected Results though dosage generally can reach at site of action, and can not produce the local concentration of harmful or poisonous in fact side effect, the amount of compound and method of administration are finally still confirmed by the doctor.

But in the whole course of treatment potion ground, continuously or off and on (for example, by the dosage of appropriate intervals) to separate realize vivo medicine-feeding.The most effective means of administration and agent method for determination of amount are known to those skilled in the art, and can be because of the used preparation of therapy, therapy purpose, to be controlled target cell and treated individual different.Single administration or many administrations can be carried out according to treatment selected dosage level of doctor and type.

Usually, the suitable dosage of lycopene is that the about 100 μ g of every kilogram of whose body weight every day are to about 250mg.If reactive compound is salt, ester, prodrug or analog, dosage is that calculate on the basis with the parent compound, so stand-by actual weight increases pro rata.

Comprise such as the lycopene compound compositions of lycopene can be individually dosed or side by side or sequentially with other treatment coupling administration, this depends on the state controlled.

The lycopene compound can be comprised in the food.For example, it can be comprised in the middle of bread, cereal, biscuit, butter, smear (as margarine), cheese, sour milk or the beverage.Other suitable food are tangible for those skilled in the art.

The lycopene compound can culinary art (as curing) preceding with the becoming phase-splitting mixing and/or after culinary art, add food of food.The data of this paper show can be incorporated the lycopene compound into food and not lose quality of food, and is being kept activity by culinary art feed product back.

Preferably, allos lycopene compound is comprised in the food.For example, above-mentioned synthetic lycopene compound or be not the natural natural lycopene compound that is present in this food.For example, the lycopene compound from fruit or vegetables can be merged in bread or cereal.

Therefore, the present invention further aspect is the food that comprises the lycopene compound of treatment metabolic function disorder, and wherein said lycopene compound is not natural this food that is present in.

Food can add the lycopene compound or strengthen food with the lycopene compound to food.In some preferred implementations, add the lycopene compound to food, described lycopene compound is used food lactalbumin preparation (" newborn lycopene ") as mentioned above.

The method of the food of preparation treatment or delay or the disorderly outbreak of prevention metabolic function also is provided, and described method comprises:

(i) provide food composition;

(ii) become phase-splitting to mix with described the lycopene compound;

(iii) with described food composition and described lycopene compound formulation feed product.

After having seen present disclosure, various further aspects of the present invention and embodiment are tangible for those skilled in the art.All it is whole and incorporate this paper into by reference for the All Files of mentioning in present specification.

Some aspect of the present invention and embodiment can illustrate in the mode of example and with reference to following figure.

Fig. 1 shows the influence of lycopene to the amount of the film of HepG2 cell and SREBP-1 in the nuclear consitution and SREBP-2.

Fig. 2 shows the immunoblotting assay of SREBP-1, SREBP-2 and LDL-R in the film of Zucker obese rat (ZFR) that lycopene was handled and thin type rat (ZLR) liver and nuclear extract.

Table 1 shows when sterol exists and do not exist, the mRNA variation of the HepG2 cell of hatching with 10 μ M lycopenes.

Table 2 shows when 100nM insulin exists and do not exist, the variation of mRNA in the rat primary hepatocyte of hatching with 10 μ M lycopenes.

Table 3 shows that the physiological parameter of the Zucker obese rat (ZFR) handled through 0.2% tomato red vegetarian meal and the thin type rat of Zucker (ZLR) (measures M ± m) for 4 times with 4 rats works.

Table 4 shows that the Zucker obese rat (ZFR) and the mRNA in the thin type rat of Zucker (ZLR) liver that handle through 0.2% tomato red vegetarian meal change.

Table 5 shows the influence of newborn lycopene LL to the human volunteer blood lipid level.

Table 6 shows is treating after two months with LL, and LL is to the influence of the average level of human volunteer blood fat.

Table 7 shows Lyc-O-Mato ^TM, LM is to the influence of human volunteer blood lipid level.

Table 8 shows is treating after two months with LM, and LM is to the influence of the average level of the blood fat of human volunteer.

Table 9 shows the influence of LL to diabetic's serum glucose concentration.

Table 10 shows the influence of LM to diabetic's serum glucose concentration.

Table 11 shows the influence of newborn lycopene LL to the serum insulin level of human volunteer.

Table 12 shows LL and the LM influence to the human volunteer body weight.

Table 13 shows the influence of Lipitor-Liprimar (Atorvastatin) to the blood lipid level of human volunteer.

Table 14 shows the antioxidation activity in vitro of the different food substrates that contains newborn lycopene (LL) through curing (surface packing and flapjack).

Table 15 shows the influence of newborn lycopene (LL) to human serum lipid metabolizing parameters.

Table 16 shows the influence of newborn lycopene (LL) to the human plasma glucose level.

Experiment

Material

Use standard molecular biological technique.(USA) extraction is from total RNA of cultured cells and rat liver for Tel-Test, Inc with the RNA-STAT-60 separation agent.(Invitrogen USA) is used to the RNA quantitative analysis to SYBR GreenqPCR reagent system.(Linco Research USA) measures plasma insulin level, and plasma glucose is measured with colorimetric method with the ELISA kit.

Unless stated otherwise, lycopene is the newborn lycopene formulations (LL) of lactalbumin combination, it is with business form (INNEOV, L ' Oreal (UK) Ltd, London) use, described form comprises in addition as the isoflavones of additive and vitamin C, perhaps use with unpack format (INDENA), its generation is by 20g tomato extract (comprising 10% newborn lycopene) and 20g lactalbumin, 50.5g microcrystalline cellulose, 5g silica, 3g polysorbate 80 and the mixing of 1.5g soybean lecithin are produced 100g breast lycopene formulations.

According to the newborn lycopene formulations of the described preparation of EP1289383.

Other general reagent obtains from Sigma (USA).

Cell is cultivated

With the density of 100 ten thousand cells/100mm culture dish, setting up individual layer human hepatocytes oncocyte is HepG2, and with it at 37 ℃ and 6% CO ₂Use and add 10% FCS and penicillin (100 units/ml) and the DMEM of streptomysin (100 μ g/ml) cultivate down.After 48 hours, use PBS to clean cell twice, and, it is gone to the DMEM that contains 5% lipoprotein-shortage serum, 50 μ M mevalonates and 50 μ M health hundred spits of fland (Compactin) (lactone form) existing or not existing under the sterol (1 μ g/ml25-hydroxy cholesterol and 10 μ g/ml cholesterol).The culture dish of half also accepts to be dissolved in the INNEOV breast lycopene of oxolane, and its final concentration is 10 μ M nearly.Half acceptance has only the aliquot of solvent in addition.After 18 hours, harvesting is used for immunoblotting assay and RNA extracts.

Primary hepatocyte is to use collagenase digestion isolated from the rat liver of non-empty stomach Spraque-Dawley rat.Under halothane anesthesia, (Gibco/BRL is USA) by each liver of portal vein situ perfusion to use liver perfusion culture medium and liver digest medium subsequently.Hepatectomize, isolating hepatocytes, and with the density of 600 ten thousand cells of each culture dish, in containing 5% hyclone and penicillin (100 units/ml) and the DMEM of streptomysin (100 μ g/ml), with its be seeded to 100mm collagen I bag quilt culture dish (Becton Dickinson Labware, USA).After three hours adhere to, described cell is gone to serum-free DMEM, it adds 100nM dexamethasone, 1nM insulin, 100nM triiodo thryonine, 100 units/ml penicillin, 100 μ g/ml streptomysins.Get rid of the method for testing living cell counting with trypan blue.The liver cell that has at least 85% living cells is used to research.After overnight incubation, existing or not existing under the 100nM insulin, add newborn lycopene (INNEOV) solution that is dissolved in oxolane, its final concentration reaches 10 μ M.The contrast culture ware is only accepted oxolane.Adding lycopene and/or insulin to liver cell after 18 hours, carry out RNA and separate.

Animal and treatment

The Zucker obese rat (fa/fa) in age in 8-9 week and its thin type littermate are used to test.Rat is raised in gregarious cage (colony cage), keeps 12 little time/12 hour dark cycle and feeding conventional food meal.After gregarious environmental adaptation 10 days, use the conventional food meal that adds 0.2% newborn lycopene (INNEOV) instead to rat.Arbitrarily feeding animals made its 24 hours can obtain described food.Every day monitoring shows through between/rat the group handled without lycopene does not have essential difference in food intake.After 8 days diet was handled, rat was put to death in halothane anesthesia down.Collect blood sample and recording body/liver weight.

RNA analyzes

Extract total RNA of cultured cell and rat liver, from each sample, get the synthetic cDNA of 2 μ g RNA, carry out the RT-PCR reaction in triplicate.Using public database Genebank is each independent gene design primer.The real-time CT value of each RNA sample of hatching with special primer all with reference to the CT value of house-keeping gene-GAPDH or 36B4 (internal reference), all shows its absolute CT value to each experiment.Observe and not exist under the sterol, the standardization CT value of each gene of HepG2 cell, and it is made as 1.00.The mRNA level of other group changes with reference to 1.00, and the representative multiple variation of this control value (1.00) relatively.

Western blotting

From institute's cultured cells with rat liver prepares nuclear extract and membrane component is used for immunoblotting assay.Allow described cell by No. 22.5 pins, smudge cells in hypotonic buffer liquid (10mM Hepes-KOH, pH 7.4).As described in addition, separate nuclear extract and membrane component from rat liver.

Clinical testing

The purpose of this clinical testing is whether under the particular formulations of not considering newborn lycopene, studying newborn lycopene self influences human metabolizing parameters.Be this test, enlisted the healthy clinically volunteer of 14 examples.12 examples wherein have a kind of dyslipidemia of or another kind of form, and 3 examples wherein also have the fasting glucose of rising in addition, and 1 example has only the fasting glucose of rising, and 1 example does not have these arbitrary metabolism symptoms.With the dosage of the HPLC lycopene of the LL of 300mg or 6mg to 7 routine volunteer's administrations, with the dosage of the HPLC lycopene of the LL of 800mg or 14mg to other 7 routine volunteer's administrations.With this product administration two months, together oral with food once a day.Test and the blood of volunteer before test and when finishing relatively.

The result

Embodiment 1

The human hepatocytes oncocyte is that HepG2 has been used to disclose the effect of newborn lycopene to the transcriptional regulatory of institute's cultured cell lipid metabolism.By table 1 as seen, in all groups of experiment, the mRNA level of basic house-keeping gene is almost constant, and from 17.54 to 17.73 do not wait.This has confirmed the RNA degraded and the health status of no drug toxicity, this cell monolayer.

By table 1 as seen, existing or not existing under the sterol, add newborn not appreciable impact of the lycopene SREBP-1mRNA level of 10 μ M.No matter whether newborn lycopene exists, and the reduction that responds to the SREBP-2mRNA of sterol in two groups is the same.

Known sterol can be blocked transcribing of SREBP-2 in institute's cultured cell, the reduction that causes the cholesterol biosynthesis of secondary to reduce and give birth to cholesterol enzyme mRNA.Table 1 has shown, the HepG2 cell is added sterol effectively reduce SREBP-2 and the biosynthetic main enzyme of cholesterol---the mRNA of FPPS, HMG-CoA Red and HMG-CoA Syn.Yet in the HepG2 cell of only hatching with 10 μ M breast lycopene, even under no sterol, the mRNA level of SREBP-2 has also obviously reduced (reducing by 42%).In the presence of newborn lycopene, the SREBP-2 level of reduction does not influence great majority and gives birth to cholesterol and give birth to the mRNA of fat.Yet in the presence of newborn lycopene, most living fat gene (FAS, ACC, HMG-CoA Syn and HMG-CoA Red) shows high slightly sterol sensitiveness.Independent newborn lycopene reduces the mRNA of SCAP, the enhancing of the sterol effect of unmatchful SCAP mRNA.

The SREBP-1 downstream gene that the newborn lycopene of unique quilt influences is INSIG-1, its mRNA 2/3 of foundation level that almost descended.Western blotting data (Fig. 1) are confirmed and have been explained mRNA result.Not existing under the sterol, in the membrane component of HepG2 cell, there is not the detected precursor forms of SREBP-1 and SREBP-2.Yet, when cutting, SREBP-1 in the presence of sterol stops, and the SREBP-1 protein content under newborn lycopene exists is obviously higher, and this has reflected that about 1.4SREBP-1mRNA raises.The Western blotting result has also confirmed the decline in the SREBP-2mRNA level seen in the HepG2 cell of hatching with newborn lycopene---in the HepG-2 cell of hatching with newborn lycopene, the amount of SREBP-2 precursor forms and mature form is obviously lower.

In the independent seminar that carries out with same operation, we have confirmed and have reproduced the inhibitory action of newborn lycopene for SCAP mRNA and INSIG-1mRNA.

But the breast lycopene reduces the main living cholesterol transcription factor SREBP-2 and the mRNA of insulin induced gene-1 (INSIG-1), is the higher a little sterol sensitiveness of giving birth to the fat gene among the HepG2 thereby give the human hepatocytes oncocyte.In a word, our result points out: newborn lycopene influences the liver specificity insulin replies of some mechanism of transcribing that lipid generates and possible human hepatocytes.

Embodiment 2

Because the gene (SREBP-1 and INSIG-1) that lycopene is regulated some insulin in the HepG2 cell has noticeable effect, select rat primary hepatocyte system and insulin to handle the physiological characteristics of operation with the newborn lycopene of further research.By table 2 as seen, handle primary rat hepatocyte with 10 μ M breast lycopene the absolute CT value of main house-keeping gene-36B4 be there is no big influence.This has confirmed not exist the acceptable viability of toxic action, primary hepatocyte and from the good quality of the RNA of described cell extraction.

The result shows that insulin optionally raises the mRNA that main liver specificity is given birth to fat transcription factor-SREBP-1c, and its CT corrected value reaches 56.5 times of foundation level.Breast lycopene self makes SREBP-1c foundation level strengthen 4 times.Simultaneously, adding newborn lycopene induces the SREBP-1c that utilizes insulin to have reduced by 2 times more than (having only 27.5 times).

Be similar to HepG-2 research, find that the existence of newborn lycopene has reduced the mRNA of main living cholesterol transcription factor-SREBP-2, particularly in the presence of insulin.In the presence of newborn lycopene, SREBP-1c reduces the level of insulin the blunt and SREBP-2 of insulin replies, looks it is that function is significant and have a metabolism consequence in primary hepatocyte.Especially, the insulin replies of main living fat mRNA-FAS, SCD-1, ACL and INSIG-1 has been eliminated in the existence of newborn lycopene fully, and described gene is known to be the target gene of SREBP.During with the treatment of newborn lycopene, these mRNA levels are corrected near control value.

Another key character of breast lycopene effect is to regulate main gluconeogenesis enzyme-PEPCK and the activity of G-6-Pase, and its mRNA level correspondingly reduces to 66.0% and 70% when using newborn lycopene separately.Insulin has almost reduced by 90% and show same ability in the presence of newborn lycopene with the mRNA of PEPCK.On the contrary, G-6-Pase is so not remarkable to replying of insulin.Interest be to use newborn lycopene to hatch and given G-6-Pase higher insulin sensitivity.Use separately insulin, this mRNA only reduces (reach control level 90%) slightly, but in the presence of insulin and newborn lycopene, descends more significantly (reach control level 62%).

The breast lycopene strengthens the insulin sensitivity of some other insulin target genes that have nothing to do with gluconeogenesis.When primary hepatocyte was exposed to insulin in the presence of newborn lycopene, insulin had more deep negative interaction to the mRNA of INSIG-2, IRS-2 and IGFBP-1.

Therefore, in the rat primary hepatocyte, newborn lycopene is reduced in living fat transcription response and the SREBP-2mRNA under the insulin existence, provides the gluconeogenesis enzyme higher insulin sensitivity.

Embodiment 3

For further the newborn lycopene of research is to the mechanism of action of hepatic metabolism, we have carried out the research of use Zucker obese rat (ZFR), and known its shown the most important characteristics of metabolic syndrome.Male and the coupling contrast (the thin type littermate of Zucker) of young (9 week big) ZFR is used to be summarized in the experiment of table 3.

Table 3 shows, even in 9 weeks when big, it is heavy that ZFR has the body weight and the liver of remarkable rising, the plasma triglyceride of rising and cholesterol.Yet plasma glucose concentration still keeps control level, although insulin level is induced by essence.Though newborn lycopene (Inneov) does not reduce the body weight of described thin type rat, be to use diet processing in 8 days of newborn lycopene to cause ZFR body weight and the heavy noticeable reduction of liver.In the control-animal (ZLR) of handling, see that also some livers heavily descend with newborn lycopene.The most significant variation occurs on plasma triglyceride and the cholesterol levels---and they both have descended 83.1% and 64.3% respectively when handling with newborn lycopene.Use the plasma insulin level of the ZFR of newborn tomato red vegetarian meal that statistical remarkable decline is also arranged.

By table 4 as seen, ZFR has the SREBP-1c transcript and the main mRNA that gives birth to the rising of lipase-FAS, SCD-1, ACL of recruitment.The breast lycopene is handled and has been reduced inducing of living fat mRNA, although this effect is a part, thereby has reflected the hyperinsulinemia that remains of the ZFR that uses newborn tomato red vegetarian meal.The Western blotting result that Fig. 2 showed shows that ZFR liver nuclear extract has remarkable higher levels of SREBP-1 protein level, and this amount does not change because of newborn lycopene processing.

Unique mRNA that drops to control value is a fatty acid synthase.Liver ACL and SCD-1mRNA have descended 2 times more than with respect to the non-processed group of ZFR.Give birth to fat mRNA these change interpretation the ZFR that handles through newborn lycopene the plasma triglyceride level decline and confirmed data shown in the embodiment 2.

In the liver of non-processing ZFR, constant SREBP-2 level is consistent with the normal value of giving birth to cholesterol enzyme (HMG-CoA-R and HMG-CoA-S), and through Western blotting data acknowledgement (see figure 2).Therefore, the hypercholesterolemia of young ZFR is absorbed owing to the LDL that reduces in the liver again, rather than the biosynthetic activation of cholesterol in the liver cell.Do not consider phenotype, newborn lycopene is handled the mRNA level (about 2 times) that has reduced the biosynthetic main transcriptional activator-SREBP-2 of cholesterol.The result from HepG2 and primary hepatocyte research shown in embodiment 1 and 2 has been supported in this discovery.Yet in diabetes and non-diabetic animal's liver, the minimizing of SREBP-2 mRNA has not same-action for HMG-CoA-R and HMG-CoA-S transcript.Obese rat shows that HMG-CoA-R and HMG-CoA-S mRNA level all significantly reduce, and thin type littermate has the level near control level.

Also have noticeable LDR-R protein induced, this immunoblotting assay by the ZFR liver of handling through newborn lycopene confirms (Fig. 1).This observation has explained that newborn tomato red vegetarian meal is to the significant normalization effect of ZFR plasma lipid level.In animal's liver, the turnover of ldl receptor is regulated by SREBP-2.In the liver of the ZFR that handles through newborn lycopene, exist the essence of SREBP-2 kernel form to raise.This increase can be explained the liver ldl receptor activation mechanism that uses newborn lycopene.

Breast tomato red vegetarian meal is for the not effect of mRNA of being responsible for cholesterol is secreted into the enzyme Cyp7-α of bile.Known Zucker obese rat is unique animal model of the low biliary cholesterol secreting rate of display abnormality.The appearance of this problem is because the reduction of CYP-7 alpha transcriptional speed in its liver.Really, by table 4 as seen, the processing that CYP7 α mRNA level descends in ZFR and do not consider to be carried out remains on almost same level.

Selecting the Zucker obese rat is because it extremely and apace forms phenotype, and it is similar to human metabolic syndrome, and causes hyperinsulinemia, body weight increase, hyperlipidemia and steatosis.All these features are all revised by newborn lycopene processing to a certain extent.

Yet by table 3 as seen, although hyperinsulinemia is arranged, 9 weeks, big ZFR still kept the blood sugar amount normal.This has illustrated, the animal that is used for this research is the gluconeogenesis that activates of development also, and it is for the key feature of diabetes B and be the obvious characteristic of another Zucker rat strain that is closely related-Zucker diabetes rat (ZDR).As the consequence of insulin sensitivity impaired (but still function is arranged), in the liver of undressed ZFR, (IGFBP-1 IRS-2) significantly reduces insulin sensitivity mRNA for PEPCK, G-6-Pase.Therefore, the insulin replies level that is kept like this of ZFR liver does not allow us to estimate in the possible body of newborn lycopene to gluconeogenesis and liver insulin sensitivity mechanism seen in the rat primary hepatocyte research (embodiment 2) to act on.

At the human liver cell oncocyte is among the HepG2 (seeing embodiment 1), and the oligogene-SREBP-2 of newborn lycopene inhibition adjusting cholesterol dynamic equilibrium transcribes.Use the cell-rat primary hepatocyte of another type to confirm this observation (seeing embodiment 2).Also reproduced the effect of newborn lycopene to SREBP-2 in the Zucker obese rat, plasma cholesterol of described rat and triglyceride levels drop to after 8 days 0.2% newborn tomato red vegetarian meal is handled and are bordering on control value.We think that newborn lycopene is not only relevant with SREBP-2 level decline in the liver to the normalization effect of plasma lipid level, also strengthens relevant with observed liver LDL-R (mRNA and albumen) level in the animal of using newborn tomato red vegetarian meal.Therefore, embodiment 1,2 and 3 has disclosed, and newborn lycopene influences cholesterol homeostasis on varying level, and described varying level refers to the adjusting of the transcribing of SREBP-2, ldl receptor expression, to the influence and the overall cholesterol synthesis rate of SCAP mRNA level.

In independent studies, find unexpectedly and reproduced, caused inhibition insulin induced gene-1 (INSIG-1) but use newborn lycopene to hatch the HepG2 cell.We think that newborn lycopene can regulate the liver specificity insulin replies at institute's cultured cell with in liver.The breast lycopene also has been observed in the rat primary hepatocyte the negative interaction of INSIG-1mRNA, particularly in the presence of insulin.Make that we are surprised to be, we also observe, and use the short-term diet of newborn lycopene to handle the body weight that reduces the Zucker obese rat and liver is heavy, plasma insulin level and cause the normalization of plasma lipid spectrum.

Therefore; INNEOV breast lycopene formulations has profound influence-reduction of plasma insulin level, plasma lipid, liver weight/weight ratio in the liver of the Zucker obese rat that newborn lycopene is handled to the various features of metabolic syndrome, gives birth to the normalization of fat genetic transcription and increasing of ldl receptor protein.

Embodiment 4

A. newborn lycopene product is to the effect of plasma lipid, glucose and insulin level

The newborn lycopene product of research the results are shown in table 5-table 11 to the effect of plasma lipid, glucose and insulin level in human volunteer.

Be this clinical testing, select the volunteer (the 8 routine male sex and 8 routine women) that 16 examples are in 45-62 year the ages.(London) LL uses Lyc-O-Mato to 12 routine administrations among them (the 6 routine male sex and 6 routine women) for INNEOV, L ' Or é al (UK) Ltd with newborn lycopene ^TM, LM (VitaHealthcare) is to 4 routine administrations among them (the 2 routine male sex and 2 routine women).One group has 2 routine diabetics in front, and there are 2 routine diabetics the back for one group.For these diabetics, except that the lipid level, also monitoring glucose concentration.Whole 16 routine patients are also monitored insulin concentration.To next dragee of 3 routine patient's administrations-whenever, twice of every day and food are obeyed altogether with LL; To next dragee of other 9 routine patient's administrations-whenever, every day three times and food are obeyed altogether with LL.LM is with three capsules and obey and administration altogether with food every day.

Before treatment and the beginning after per two weeks from blood samples of patients, collect blood serum sample.Use the treatment of two kinds of lycopene products to continue two months.

LL and LM the results are shown in table 5, table 6, table 7 and table 8 to the influence of these volunteers' lipid parameter.These results show, every day 2 or 3 dragees LL administration (table 5 and table 6) or every day 3 capsules the administration (table 7 and table 8) of LM caused the decline of the concentration of T-CHOL and triglycerides.The administration of LL also causes the remarkable decline (p＜0.05) of CH-LDL concentration separately.Other lipid parameter of volunteer is not by appreciable impact.

Notice that interestingly the level of these lipids is high more before this treatment, the decline of its concentration is just deep more in the observed serum.For example, for No. 9 patient, with LL treatment only after two weeks, total cholesterol level is just reduced to 158mg/dL from 247mg/dL, and almost 36% (table 5) descended.

Use the relatively demonstration of the effect of the different prepared product administrations of lycopene, for the level of serum total cholesterol, LDL cholesterol and triglycerides, LL has the effect more deep than LM.

In addition, also importantly, it is the reduction (table 9 and table 10) of serum glucose concentration among the patient of biological available lycopene prepared product LL or the LM preparation serum glucose that causes having elevated levels that use can absorb.

Observe interestingly, do not influence non-hyperglycemia patients serum's glucose physiological level with LL or LM administration, but can be with its reduction or normalization (runic number) in the hyperglycemia patient.

Hyperglycemia patient has those of the serum glucose concentration that is higher than 6mmol/1.

Use the relatively demonstration of the effect of the different prepared product administrations of lycopene, LL can on average reduce by 28% with hyperglycemia patient's plasma glucose concentration, and LM reduces by 16% with this parameter only.

For insulin level, also observe the similar effect of LL.In the patient that insulin level raises, the LL administration in two weeks causes its normalization (runic number).Yet this prepared product does not influence the insulin concentration (table 11) among the patient of the insulin with normal physiological level.

B. lycopene product is to the effect of human body weight

Second clinical experimental study effect of lycopene product to body weight, this test relates to 18 routine volunteers, they are the 10 routine male sex and 8 routine women, the age was at 45 years old to 62 years old.Lacto-Lycopene with 1 dragee ^TM(London) LL is to 9 routine volunteers (the 5 routine male sex and 4 routine women) administration for INNEOV, L ' Or é al (UK) Ltd, and every day three times and food are obeyed altogether.Lyc-O-Mato with 1 capsule ^TM, LM (Vita Healthcare) is to other 9 examples (the 5 routine male sex and 4 routine women) administration, and every day three times and food are obeyed altogether.

Test duration two months is weighed to the volunteer before test and after the test.Result of the test is shown in table 12.

LL and LM administration caused having respectively in each treatment group 5 examples and 3 routine volunteer's body weight to descend in two months.Yet, many 3 times of the TBW suppression ratio LM group of whole LL group, i.e. the former 15kg and latter 5kg.On an average, LL organizes everyone loss of weight 1.7kg, and LM organizes everyone loss of weight 0.6kg.Notice that interestingly overweight volunteer is more remarkable from this benefit on of LL.4/5ths loss of weight 2kg to 5kg among them.In the LM group, had only 1 routine loss of weight among the 5 routine overweight persons.

Therefore, the lycopene treatment has reduced the basic Clinical symptoms of metabolic syndrome (hypercholesterolemia, hypertriglyceridemia and hyperglycemia) among the patient.

These clinical datas have been supported observed experimental result in experiment in vitro and zoopery, and illustrated and to have absorbed and biological can to utilize the newborn lycopene prepared product of form can be effective therapeutic product, it can be used for treating metabolic syndrome, insulin resistance (comprising the severe insulin resistance syndrome), glucose tolerance, PCOS (PCOS), hypertension, steatosis, chronic hepatitis, the fibrillatable of liver, sclerosis, and is used for body weight control and is used for the treatment of obesity.

C. his spit of fland is to the effect of blood fat

In human volunteer, studied the effect of Lipitor-Liprimar (Atorvastatin), the results are shown in table 13 blood lipid level.

Be this clinical testing, selected 10 routine volunteers, they are the 5 routine male sex and 5 routine women, and the age is in 45-55 year.To each patient's oral administration, obey administration two months with 40mg Lipitor-Liprimar altogether every day with food.

From blood samples of patients, collect blood serum sample before treatment and after the bimestrial treatment.

Lipitor-Liprimar is to the table 13 that the results are shown in of the influence of blood lipid level among these volunteers.Digital proof shown in this table, this HMG CoA of Lipitor-Liprimar reductase inhibitor has reduced CH-LDL (the former 35% and the latter 13.3%) better than LL.Yet, compare his spit of fland, newborn lycopene has dropped to a lower level with T-CHOL and triglycerides: for Lipitor, T-CHOL (CH) has descended 17%, and is 27% to LL; For Lipitor, triglycerides (TG) has descended 22%, and is 34% (table 6 and table 13) to LL.

Just do not reducing on the blood lipid level ineffectively inadequately, known his spit of fland also has some side effects to 0.5% to 2.0% patient.For example, Ta Ting can cause transaminase level rising or myalgia, tenderness or unable.In some patients, these symptoms can be followed heating or influenza-like symptom, stomachache or unaccountable fatigue (unexplained fatigue).In addition, liver diseases and myositis belong to the contraindication of taking his spit of fland, and do not answer Kai Tating in the pregnancy period.On the other hand, long term administration breast lycopene prepared product does not have known contraindication or side effect.

In conjunction with data shown in above, the newborn lycopene formulations of this explanation such as LL can be used as the substitute in his spit of fland in treatment comprises the various symptoms of metabolic syndrome, insulin resistance, GI, hypertension, PCOS, obesity, steatosis, chronic hepatitis and cirrhosis.

In addition, such as the newborn lycopene formulations of LL can with his spit of fland coupling, this can reduce the latter's therapeutic dosage.Contraindication in his spit of fland is reduced for this and the side effect that it is possible minimizes.

The preparation that D is other

In human volunteer, studied of the effect of the newborn lycopene formulations (INDENA) of separation, the results are shown in table 15 and table 16 blood lipid level.

These results show that newborn lycopene formulations (INDENA) has reduced blood lipid level, but to not significantly effect of glucose level in the human plasma.Yet it makes the concentration normalization of T-CHOL and triglycerides in the serum of described individuality of T-CHOL with elevated levels and triglycerides.The level of these parameters is high more before the treatment, and observed reduction is just remarkable more.For example, above individual average 94mg/dL of reduction or the reduction of T-CHOL 250mg/dL surpasses 35%.

Initial concentration 250mg/dL is following but still be higher than the individual average 54mg/dL of reduction of 200mg/dL normal level or reduce by 25%.For triglycerides, observe similar type.Even in the individuality of these lipids, observed the reduction-T-CHOL 15% of two kinds of parameters and triglycerides 20% with physiology normal level.

For the concentration of CH-HDL and CH-LDL, observe so not deep effect.In the 5 routine individualities that the former level reduces, 4 examples have the described lipid levels of normalization after treatment.In the initial 4 routine individualities that raise of CH-LDL level, this parameter of having only 2 examples is by normalization.

Embodiment 5

Incorporate newborn lycopene into food matrix

The prescription of piece of bread is:

Yeast-3/4 tea spoonful, high muscle light flour-400g ,-1 of sugar, butter-15g, milk powder-1 tea spoon, salt-1 tea spoonful, 280ml has dissolved (INNEOV) water of dragee of newborn lycopene (LL).

Cure four: one (contrast) do not contain LL, second LL that contains 5 dragees, the 3rd LL that contains 10 dragees, the 4th LL that contains 20 dragees.

Test

The 5 routine volunteers that cover the described bread of eye test do not find to have any different on quality or fragrance between all these four.In addition, tested the antioxidation activity of bread.

Abide by the operation of in manufacturers instruction, stating, use AtheroAbzyme ^TM(CTL UK) finishes it ELISA kit.

The bread that comprises newborn lycopene has antioxidation activity, and contrast bread does not have (table 14).Embodiment 6

Incorporate lycopene into food matrix

The prescription of 4 a collection of flapjacks is:

Self-raising flour-175g, yeast powder-1 tea spoonful, pugil salt, castor sugar-20g, salt-free butter-37g, 90ml have dissolved the milk of the LL of INNEOV dragee form.

Cure four batches of flapjacks: a collection of (contrast) do not contain LL, second crowd of LL that contains 0.5 dragee of every flapjack, the 3rd crowd of LL that contains 1 dragee of every flapjack, the 4th crowd of LL that contains 2 dragees of every flapjack.

Test

The 5 routine volunteers that cover these flapjacks of eye test do not find to have any different on quality or fragrance between all these four batches.In addition, tested the antioxidation activity of these flapjacks.Abide by the operation of in manufacturers instruction, stating, use AtheroAbzyme ^TM(CTL UK) finishes it ELISA kit.The flapjack that comprises newborn lycopene has antioxidation activity, and the contrast flapjack does not have (table 14).

Abbreviation: GAPDH-glyceraldehyde-3-phosphate dehydrogenase; SREBP-1-sterol regulating element is in conjunction with albumen-1; SREBP-2-sterol regulating element is in conjunction with albumen-2; FPPS-farnesyl diphosphate synthase; HMG-CoA Syn-HMG-CoA synthase; HMG-CoA Red-HMG-CoA reductase; SCD-1-stearoyl-coacetylase-desaturase-1; FAS-fatty acid synthase; ACC-acetyl-CoA carboxylase; SCAP-sterol cutting activator protein; But INSIG-1-insulin induced gene-1; But INSIG-2-insulin induced gene-2

Table 1

Abbreviation: 36B4-acid ribosomes phosphoprotein; SREBP-1-sterol regulating element is in conjunction with albumen-1; SREBP-2-sterol regulating element is in conjunction with albumen-2; FAS-fatty acid synthase; SCD-1-stearoyl-coacetylase-desaturase-1; ACL-ATP-citrate lyase; PEPCK-phosphoenolpy ruvate carboxy kinase; IGFBP-1-insulin-like growth factor binding protein; G-6-Pase-G-6-Pase; IRS-2-IRS 2; But INSIG-1-insulin induced gene-1; But INSIG-2-insulin induced gene-2; Insulin-R-insulin receptor

Table 2

Table 3

Abbreviation: 36B4-acid ribosomes phosphoprotein; SREBP-1-sterol regulating element is in conjunction with albumen-1; SREBP-2-sterol regulating element is in conjunction with albumen-2; FAS-fatty acid synthase; SCD-1-stearoyl-coacetylase-desaturase-1; ACL-ATP-citrate lyase; PEPCK-phosphoenolpy ruvate carboxy kinase; IGFBP-1-insulin-like growth factor binding protein; G-6-Pase-G-6-Pase; IRS-1-substrate 1; IRS-2-IRS 2; But INSIG-2-insulin induced gene-2; LDL-R-LDL receptor; CYP-7 α-7 α-hydroxylase; But INSIG-1-insulin induced gene-1; HMG-CoA-S-HMG-coacetylase synthase; HMG-CoA-R-HMG-CoA-reductase; But INSIG-1-insulin induced gene; But INSIG-1-insulin induced gene-1

Table 4

^*＂ INNEOV ＂ dragee number (Lacto-Lycopene of each dragee 20mg)

^*CH-T-CHOL, TG-triglycerides

Table 5

^*Statistically evident

Table 6

^*Every day is to Drug Capsule number (each capsule 15mg lycopene)

^*CH-T-CHOL, TG-triglycerides

Table 7

^*Statistically evident

Table 8

Table 9

Table 10

Table 11

Table 12

^*Every day oral Lipitor 40mg

Table 13

Table 14

The lipid parameter runic of unusual too high or too low level

^*Patient number

Table 15

Table 16

Claims (85)

1. the method for treatment metabolic function disorder comprises and will treat the lycopene compound administration of effective dose in its individuality of needs.

2. the method for claim 1, wherein said lycopene compound is a lycopene.

3. method as claimed in claim 1 or 2, wherein said metabolic function disorder are selected from obesity, insulin resistance, glucose tolerance reduction, PCOS, hypertension, steatosis, chronic hepatitis, liver fibrosis, sclerosis and metabolic syndrome.

4. as each the described method in the claim 1 to 3, wherein prepare described lycopene compound with lactalbumin.

5. as each the described method in the claim 1 to 3, wherein with the described lycopene compound of oil preparation.

6. as each the described method in the claim 1 to 5, wherein prepare described lycopene compound with one or more isoflavones.

7. as each the described method in the claim 1 to 6, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

8. as each the described method in the claim 1 to 7, wherein said lycopene compound is comprised in the food.

9. method as claimed in claim 8, wherein said food are bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

10. as each the described method in the claim 1 to 9, wherein said lycopene compound is used for and his spit of fland administering drug combinations.

11., comprise with described lycopene compound and his spit of fland coupling administration as each the described method in the claim 1 to 8.

12. the purposes of lycopene compound in the medicine of preparation treatment metabolic function disorder.

13. purposes as claimed in claim 12, wherein said lycopene compound is a lycopene.

14. as claim 12 or 13 described purposes, wherein said metabolic function disorder is selected from obesity, insulin resistance, glucose tolerance reduction, PCOS, hypertension, steatosis, chronic hepatitis, liver fibrosis, sclerosis and metabolic syndrome.

15., wherein prepare described lycopene compound with lactalbumin as each the described purposes in the claim 12 to 14.

16. as each the described purposes in the claim 12 to 14, wherein with the described lycopene compound of oil preparation.

17., wherein prepare described lycopene compound with one or more isoflavones as each the described purposes in the claim 12 to 16.

18. as each the described purposes in the claim 12 to 17, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

19. as each the described purposes in the claim 12 to 17, wherein said lycopene is comprised in the food.

20. purposes as claimed in claim 18, wherein said food are bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

21. as each the described purposes in the claim 12 to 20, wherein said medicine is used for and his spit of fland administration.

22. as each the described purposes in the claim 12 to 21, wherein said medicine also comprises his spit of fland.

23. the methods of treatment of the state relevant with the metabolic function disorder comprises and will treat the lycopene compound administration of effective dose in its individuality of needs.

24. method as claimed in claim 23, wherein said lycopene compound is a lycopene.

25., wherein be selected from blood pressure rising, hypertriglyceridemia, hypercholesterolemia, high LDL cholesterol and/or low HDL cholesterol and microalbuminuria with the disorderly relevant state of metabolic function as claim 23 or 24 described methods.

26., wherein prepare described lycopene compound with lactalbumin as each the described method in the claim 23 to 25.

27. as each the described method in the claim 23 to 25, wherein with the described lycopene compound of oil preparation.

28., wherein prepare described lycopene compound with isoflavones as each the described method in the claim 23 to 25.

29. as each the described method in the claim 24 to 28, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

30. as each the described method in the claim 24 to 29, wherein said lycopene is comprised in the food.

31. method as claimed in claim 30, wherein said food are bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

32. as each the described method in the claim 24 to 31, wherein said lycopene compound is used for and his spit of fland administration.

33., comprise and the described lycopene compound of his spit of fland coupling administration as each the described method in the claim 24 to 32.

34. the lycopene compound is used for the treatment of purposes in the medicine with the disorderly relevant state of metabolic function in preparation.

35. purposes as claimed in claim 34, wherein said lycopene compound is a lycopene.

36. as claim 34 or 35 described purposes, the wherein said state relevant with the metabolic function disorder is selected from blood pressure rising, hypertriglyceridemia, hypercholesterolemia, high LDL cholesterol and/or low HDL cholesterol and microalbuminuria.

37., wherein prepare described lycopene compound with lactalbumin as each the described purposes in the claim 34 to 36.

38. as each the described purposes in the claim 34 to 36, wherein with the described lycopene compound of oil preparation.

39., wherein prepare described lycopene compound with isoflavones as each the described purposes in the claim 34 to 36.

40. as each the described purposes in the claim 35 to 39, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

41. each the described purposes as in the claim 35 to 40 wherein is formulated in described lycopene in the food.

42. purposes as claimed in claim 41, wherein said food are bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

43. as each the described purposes in the claim 35 to 42, wherein said medicine is used for and his spit of fland administration.

44. as each the described purposes in the claim 33 to 39, wherein said medicine also comprises his spit of fland.

45. kit comprises the lycopene compound of uniting use of treatment metabolic function disorder and Ta Ting.

46. kit as claimed in claim 45, wherein said lycopene compound is a lycopene.

47. as claim 45 or 46 described kits, wherein said metabolic function disorder is selected from obesity, insulin resistance, glucose tolerance reduction, PCOS, hypertension, steatosis, chronic hepatitis, liver fibrosis, sclerosis and metabolic syndrome.

48., wherein prepare described lycopene compound with lactalbumin as each the described kit in the claim 45 to 47.

49. as each the described kit in the claim 45 to 47, wherein with the described lycopene compound of oil preparation.

50., wherein prepare described lycopene compound with isoflavones as each the described kit in the claim 45 to 47.

51. as each the described kit in the claim 46 to 50, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

52. kit comprises the lycopene compound and the Ta Ting that unite use that treat the state relevant with the metabolic function disorder.

53. kit as claimed in claim 52, wherein said lycopene compound is a lycopene.

54. as claim 52 or 53 described kits, wherein said state is selected from obesity, insulin resistance, glucose tolerance reduction, PCOS, hypertension, steatosis, chronic hepatitis, liver fibrosis, sclerosis and metabolic syndrome.

55., wherein prepare described lycopene compound with lactalbumin as each the described kit in the claim 52 to 54.

56. as each the described kit in the claim 52 to 54, wherein with the described lycopene compound of oil preparation.

57., wherein prepare described lycopene compound with isoflavones as each the described kit in the claim 52 to 54.

58. as each the described kit in the claim 52 to 57, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

59. the preparation that is combined in of lycopene compound and Ta Ting is used for the treatment of purposes in the medicine of metabolic function disorder.

60. purposes as claimed in claim 59, wherein said lycopene compound is a lycopene.

61. as claim 59 or 60 described purposes, the wherein said state relevant with the metabolic function disorder is selected from blood pressure rising, hypertriglyceridemia, hypercholesterolemia, high LDL cholesterol and/or low HDL cholesterol and microalbuminuria.

62., wherein prepare described lycopene compound with lactalbumin as each the described purposes in the claim 59 to 61.

63. as each the described purposes in the claim 59 to 61, wherein with the described lycopene compound of oil preparation.

64., wherein prepare described lycopene compound with one or more isoflavones as each the described purposes in the claim 59 to 61.

65. as each the described purposes in the claim 61 to 64, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

66. each the described purposes as in the claim 60 to 65 wherein is formulated in described lycopene in the food.

67. as the described purposes of claim 66, wherein said food is bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

68. the preparation that is combined in of lycopene compound and Ta Ting is used for the treatment of purposes in the medicine with the disorderly relevant state of metabolic function.

69. as the described purposes of claim 68, wherein said lycopene compound is a lycopene.

70. as claim 68 or 69 described purposes, the wherein said state relevant with the metabolic function disorder is selected from blood pressure rising, hypertriglyceridemia, hypercholesterolemia, high LDL cholesterol and/or low HDL cholesterol and microalbuminuria.

71., wherein prepare described lycopene compound with lactalbumin as each the described purposes in the claim 68 to 70.

72. as each the described purposes in the claim 68 to 70, wherein with the described lycopene compound of oil preparation.

73., wherein prepare described lycopene compound with one or more isoflavones as each the described purposes in the claim 68 to 70.

74. as each the described purposes in the claim 69 to 73, wherein said lycopene compound and pharmaceutically acceptable excipient are comprised in the pharmaceutical composition.

75. each the described purposes as in the claim 69 to 74 wherein is formulated in described lycopene in the food.

76. as the described purposes of claim 75, wherein said food is bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

77. comprise the food of the lycopene compound of treatment metabolic function disorder, wherein said lycopene compound and non-natural are present in described food.

78. as the described food of claim 77, wherein said food is bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

79. as claim 77 or 78 described food, wherein said lycopene compound is a lycopene.

80. as each the described food in the claim 77 to 79, wherein said metabolic function disorder is selected from obesity, insulin resistance, glucose tolerance reduction, PCOS, hypertension, steatosis, chronic hepatitis, liver fibrosis, sclerosis and metabolic syndrome.

81. as each the described food in the claim 77 to 80, described food also comprises his spit of fland.

82. the method for the food of preparation treatment metabolic function disorder comprises:

(i) provide food composition;

(ii) the lycopene compound is mixed with described composition;

(iii) described composition and described lycopene compound are formulated in the food.

83. as the described method of claim 82, wherein said food is bread, cereal, biscuit, butter, smear, cheese, sour milk or beverage.

84. as claim 82 or 83 described methods, wherein said lycopene compound is a lycopene.

85. as each the described method in the claim 82 to 84, wherein said metabolic function disorder is selected from obesity, insulin resistance, glucose tolerance reduction, PCOS, hypertension, steatosis, chronic hepatitis, liver fibrosis, sclerosis and metabolic syndrome.

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