CN109528748A - Application of the cyclic adenosine monophosphate salt in preparation external application fat-eliminating slimming product - Google Patents
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Abstract
The present invention relates to application of the cyclic adenosine monophosphate salt in preparation external application fat-eliminating slimming product, belong to pharmaceutical technology field.Cyclic adenosine monophosphate salt of the invention can be directly entered activated protein kinase A in fat cell, and then activate lipase-catalyzed lipolysis into free fatty acid and glycerol, achieve the purpose that the rouge that disappears.Cyclic adenosine monophosphate salt of the invention can reduce the fat mass in fat cell, can be developed into the product of external application fat-eliminating slimming.
Description
Technical field
The present invention relates to application of the cyclic adenosine monophosphate salt in preparation external application fat-eliminating slimming product, belong to medical science neck
Domain.
Background technique
According to the difference at body fat Tissue distribution position, obesity can be divided into adiposis universalis and localised adiposities (upper body fertilizer
Fat and lower part of the body obesity, Central obesity) two major classes.Adiposis universalis has become that global, serious society is public to be defended at present
Raw problem.The statistical result of World Health Organization's publication shows that global at least 1,000,000,000 adults at present are overweight, and 300,000,000 people are fat,
Population obesity has seriously threatened global evolution.And although localised adiposities generally not will cause serious health problem,
It is a more common social concern, it refers to the obesity that a certain partial fat accumulation of body is excessive and occurs, with abdomen
The positions such as (beer belly/beer belly), arm (butterfly arm), thigh (elephant leg) and buttocks are most commonly seen.Currently, as people are raw
The flat continuous improvement of running water, diet are abundant, overnutrition, almost all people it is all more or less there are localised adiposities,
Personal overall image and beauty are seriously affected, to many artificial at great puzzlement.
Current local method for lowering fat and weight-reducing mainly includes liposuction, injection fat melting needle etc..But liposuction may result in skin
Skin recess, relaxation, or even there are the serious side effects such as subcutaneous extvavasated blood and bleeding.And the fat melting complicated component of fat melting needle and unknown
Really, the serious problems of swelling, pain or even infection and necrosis are easy to appear, by the world it is multinational include China prohibite use.
And the dazzling rouge that disappears, weight reducing and the body-shaping slimming product overwhelming majority belongs to invalid product on the market.
To sum up, safely and effectively local weight-reducing product and technology be there is no on the market at present, exploitation is safely, conveniently and efficiently
Local fat-eliminating slimming product undoubtedly has huge market potential.
The molecular formula of cyclic adenosine monophosphate salt is C10H12N5XO6P, structural formula are indicated by following formula I:
Cyclic adenosine monophosphate salt is used for angina pectoris, myocardial infarction, myocarditis and cardiogenic shock.To improvement rheumatic heart disease
Palpitaition, the symptoms such as out of breath, uncomfortable in chest have certain effect.Curative effect can be improved to acute leukemia combination chemotherapy, also can be used for urgency
The inducer remission of property leukaemia.In addition, also having certain curative effect to senile chronic bronchitis, various hepatitis and psoriasis.But
The fat-eliminating slimming effect of cyclic adenosine monophosphate salt is had not been reported at present.
Summary of the invention
Cyclic adenosine monophosphate salt is provided outside preparation it is an object of the invention to overcome above-mentioned the deficiencies in the prior art place
With the application in fat-eliminating slimming product, cyclic adenosine monophosphate salt of the invention can be directly entered activated protein kinase in fat cell
A, and then lipase-catalyzed lipolysis is activated into free fatty acid and glycerol, reach the rouge purpose that disappears.
To achieve the above object, the technical scheme adopted by the invention is as follows: cyclic adenosine monophosphate salt preparation external application fat-eliminating slimming
Application in product, the cyclic adenosine monophosphate salt are compound of formula I, wherein X indicates salt ion
As the preferred embodiment of application of the present invention, the cyclic adenosine monophosphate salt be cyclic adenosine monophosphate and metal from
The pharmaceutically acceptable salt that son is formed.
As the preferred embodiment of application of the present invention, the pharmaceutically acceptable salt is sodium salt.
As the preferred embodiment of application of the present invention, the product is drug, health care product or cosmetics.
As the preferred embodiment of application of the present invention, the external application fat-eliminating slimming product is to reduce in fat cell
The product of fat mass.
Second aspect, the present invention provides a kind of composition of external application fat-eliminating slimming, the composition contains cycli phosphate gland
Glycosides salt.
As the preferred embodiment of composition of the present invention, the composition also contains ivy extract, the moon is shown in
At least one of careless extract, cocoa extract, L-carnitine.
The third aspect, the present invention provides a kind of product of external application fat-eliminating slimming, the product includes above-mentioned composition.
As the preferred embodiment of product of the present invention, the product is drug, health care product or cosmetics.
As the preferred embodiment of product of the present invention, the dosage form of the product be paste, paste, paint, patch,
Gelling agent, liniment or spray.
Compared with prior art, the invention has the benefit that cyclic adenosine monophosphate salt of the invention can be directly entered rouge
Activated protein kinase A in fat cell, and then lipase-catalyzed lipolysis is activated into free fatty acid and glycerol, reach the rouge that disappears
Purpose；Cyclic adenosine monophosphate salt of the invention can reduce the fat mass in fat cell, can be developed into the production of external application fat-eliminating slimming
Product.
Detailed description of the invention
Fig. 1 is influence statistical chart of the cyclic adenosine monophosphate salt to Fat Accumulation.
Fig. 2 is influence statistical chart of the ivy extract to Fat Accumulation.
Fig. 3 is influence statistical chart of the Radix Oenotherae erythrosepalae extract to Fat Accumulation.
Fig. 4 is influence statistical chart of the cocoa extract to Fat Accumulation.
Fig. 5 is influence statistical chart of the pharmaceutical composition to Fat Accumulation.
Fig. 6 is pharmaceutical composition to the influence statistical chart of obese mouse weight and abdominal subcutaneous fat thickness, wherein A is
The influence statistical chart of each group mouse weight, B are the influence statistical chart of each group mouse subcutaneous fat thickness.
Specific embodiment
Purposes, technical schemes and advantages in order to better illustrate the present invention, below in conjunction with the drawings and specific embodiments pair
The present invention is described further.
Embodiment 1
1, the preparation of cyclic adenosine monophosphate salt
Using adenosine as raw material, phosphorus oxychloride is phosphorus phthalein reagent, makees solvent with tricresyl phosphate second vinegar, generates adenosine under low temperature
This intermediate is separated with reaction system, dissolves in solvent appropriate, be slowly added to KOH with stirring by 5' dichloro phosphoric acid vinegar
Water and acetonitrile solution in, highly basic is cyclized to obtain 3', 5'- cyclic adenosine monophosphate.By weight for 1:0.25 weigh adenosine cyclophosphate with
And sodium bicarbonate, the sodium bicarbonate of recipe quantity is dissolved in 60% and matches the water for injection of liquid measure, then recipe quantity is added while stirring
Adenosine cyclophosphate, make it completely dissolved；The medicinal carbon of 0.01%g/mL is added, it is stirring and adsorbing 30 minutes, micro- through 0.45 μm
Hole membrane filtration carbon removal, filtrate add to the full amount of water for injection, and being sufficiently stirred is uniformly mixed solution, prepared solution
End-filtration degerming is carried out through 0.22 μm of miillpore filter, obtains filtrate；Filtrate is encased in injection bottle, low temperature cold freeze-drying
It is dry, obtain adenosine cyclophosphate sodium salt freeze-dried powder.
2, the preparation of ivy extract, Radix Oenotherae erythrosepalae extract, cocoa extract
The preparation of ivy extract:
A part of Changchun cane leaves are highly crushed in pulverizer, ensure that the full-size of fragment is 2x2mm with protection sieve.
The water section in 6 parts of extractants (30% (m/m) ethyl alcohol) is added into the sample of crushing.It is mixed frequently, this is mixed
Object is closed to ferment 60 minutes in 30 DEG C.Then 96% ethanolic moiety in 6 portions of extractants is added, by stirring mixture homogenization.
After pre-swollen 6 hours, eluent is isolated, the medicinal herbs that left behind are shifted to an earlier date into liquid with remaining 6 parts and are percolated.By merging
Eluent filters again, to remove medicinal herbs little particle, after being allowed to homogenizing, under 55 DEG C and 150mbar pass through thin film evaporation by its
It is dried to obtain concentrate.Concentrate is homogenized, is then dried in 45 to 60 DEG C by spray drying, obtains dry Changchun
Boisiana extract.
Radix Oenotherae erythrosepalae extract preparation:
Oenothera biennis is ground into coarse powder, 10~80 meshes is crossed, is placed in extraction kettle and is extracted, extracting pressure is 7.0~
45Mpa, 30~80 DEG C of extraction temperature, extraction time 30min~8h, the supercritical CO dissolved with extract2Into in separating still
Parsing separation is carried out, 4.0~7.0Mpa of pressure is parsed, 20~80 DEG C of desorption temperature, parses time 30min~8h, obtain oenothera biennis
Extract.
The preparation of cocoa extract:
500g cocoa bean is weighed, is crushed, is dispersed in after cocoa bean is crushed and is stirring evenly and then adding into 0.5g in 1500g water and mixes
Synthase (cellulase: pectase: protease=3:1:1).Said mixture is heated to 40 DEG C, 1h is stirred to react and is digested
Liquid.An extracting solution is obtained by filtration after enzymatic hydrolysis.Filter residue is the cocoa power after enzymatic hydrolysis.All enzymatic hydrolysis cocoa powers are added to
In 95% ethyl alcohol of 500g, mixed liquor is heated to 65 DEG C, is stirred at reflux carry out second extraction, while going out to remaining mixed enzyme
Living, the second extraction time is 1.5h.The filtrate for merging the extraction of two steps carries out extracting solution pre- dense using ultrafiltration membrane separating device
Contracting purification.Select aperture for 0.03 μm of membrane material, operating pressure 1MPa.The extracting solution of retention is concentrated into using cryogenic vacuum
Required concentration.
Embodiment 2
1, test method
3T3-L1 PECTORAL LIMB SKELETON induces differentiation into fat cell.By the fibroblast system 3T3-L1 of rat with 5 ×
104The density inoculating cell of/milliliter is containing 10% fetal calf serum (FBS, GIBCO, Life in the culture plate in 12 holes
Technology culture in DMEM in high glucose culture medium (GIBCO, Life technology)).It is long to 70% degrees of fusion to cell
When, culture medium is changed into containing 1 mcg/ml insulin, 0.25 micromoles per liter dexamethasone (dexamethasone) and 0.5 milli
(containing 10%FBS) in the new DMEM culture medium of mol/L 3-isobutyl-1-methylxanthine (IBMX), induction is broken up；It will after 2 days
Culture medium changes into the new DMEM culture medium (containing 10%FBS) of 1 mcg/ml insulin；Culture medium is changed into normally after 2 days
DMEM culture medium (containing 10%FBS) is simultaneously observed, until forming fat cell.
Experimental method 1: pass through the accumulation situation of oil red O staining method and microplate reader detection fat.It is bought using from sigma
Oil red O, the solution of 10 mg/mls is configured to isopropanol, is kept in dark place after filtering.3T3-L1 fat cell phosphate
Buffer (PBS) rinse 2 times, 10% formaldehyde fixes 30 minutes or more；With oil red: deionized water=3:2 dilution, filter paper filtering,
It is placed at room temperature for 10min；Cell dyeing 10min or so removes extra dyestuff then with 75% alcohol/60% isopropyl alcohol；
It micro- sem observation and takes pictures.It is dissolved after taking pictures using 100% isopropanol, harvests lysate, examined at 510nm wavelength through microplate reader
Survey light absorption value.Test group is the group that each component is used alone or in combination, and control group is the group of no added any component.
Experimental method 2: the foundation and grouping of fat animal model are administered: animal model preparation and grouping: 4 week old C57/
BJ6L male mice is fed normal diet 7 days under experimental enviroment, is weighed, the mouse high lipid food for selecting weight almost the same
It feeds 8 weeks, is more than the mouse with normal diet raising mouse weight 20% as obesity mice using weight, is included in experimental group:
Fat control group, combinational drug therapy group.Mouse divides cage to feed, ad lib drinking-water；Naturally daylighting round the clock, room temperature 18~25
℃；Drug is used for the abdomen of mouse, unhairing before smearing by the daily timing of combinational drug therapy group in a manner of smearing.Every 2~3 days
Change food, the next day change water；Relative humidity 50% or so；Experiment periods are 30 days.Groups of animals is fed with high lipid food.Treatment is completed
After measure weight, skin of abdomen carries out H&E dyeing and takes pictures to calculate the thickness of subcutaneous fat after fixed embedding.Wherein, commonly
Feed are as follows: moisture: 8.4%, crude protein 25.8%, crude fat 6.53%, coarse ash 6.2%, crude fibre 4.12%, nitrogen-free leaching
Object 46.8%, calcium 1.26%, phosphorus 0.89%, lysine 1.36%.High lipid food are as follows: moisture: 8.6%, crude protein 18.8%, slightly
Fat: 18.83%, coarse ash 5.2%, crude fibre 3.98%, nitrogen-free extracts 45.2%, calcium 1.24%, phosphorus 0.83%, bad ammonia
Acid 1.38%.
2, test result
(1) cyclic adenosine monophosphate sodium salt promotes the effect of lipolysis
In order to measure the effect that cyclic adenosine monophosphate sodium salt promotes the lipolysis in fat cell, using in Preparatory work of experiment
The 3T3-L1 fat cell of differentiation.Cell culture medium is changed into the new DMEM culture medium of 10 mg/ml cyclic adenosine monophosphate sodium salts
(containing 10%FBS) is then detected accumulation situation fatty in fat cell by experimental method 1.As a result as shown in Figure 1, its
In, P < 0.001 * * P < 0.01, * * *.
For Fig. 1 the results show that compared with the control group, the fat mass in cyclic adenosine monophosphate sodium salt treated fat cell is obvious
It reduces, and lipolytic effect is better than the known positive comparative control caffeine with the effect that reduces fat.
(2) the effect of ivy extract inspires lipolysis
In order to measure the effect that ivy extract promotes the lipolysis in fat cell, divide using in Preparatory work of experiment
The 3T3-L1 fat cell of change.Cell culture medium is changed the new DMEM culture medium of 10 mg/litre ivy extracts into (containing 10%
FBS), then, accumulation situation fatty in fat cell is detected by experimental method 1.As a result as shown in Figure 2, wherein * P <
0.05, * P < 0.001 * P < 0.01, * * *.
Fig. 2 is the results show that compared with the control group, the neutral fat in fat cell after ivy extract-treated is bright
It is aobvious to reduce.
(3) Radix Oenotherae erythrosepalae extract inspires the effect of lipolysis
In order to measure the effect that Radix Oenotherae erythrosepalae extract promotes the lipolysis in fat cell, divide using in Preparatory work of experiment
The 3T3-L1 fat cell of change.Cell culture medium is changed the new DMEM culture medium of 10 mg/litre Radix Oenotherae erythrosepalae extracts into (containing 10%
FBS), then, accumulation situation fatty in fat cell is detected by experimental method 1.As a result as shown in Figure 3, wherein * P <
0.05, * P < 0.001 * *.
Fig. 3 is the results show that compared with the control group, and fat mass obviously drops in Radix Oenotherae erythrosepalae extract treated fat cell
It is low.
(4) cocoa extract inspires the effect of lipolysis
In order to measure the effect that cocoa extract promotes the lipolysis in fat cell, break up using in Preparatory work of experiment
3T3-L1 fat cell.Cell culture medium is changed the new DMEM culture medium of 10 mg/litre cocoa extracts into (containing 10%
FBS), then, accumulation situation fatty in fat cell is detected by experimental method 1.As a result as shown in Figure 4, wherein * P <
0.05, * P < 0.001 * P < 0.01, * * *.
Fig. 4 is the results show that compared with the control group, and fat mass is substantially reduced in cocoa extract treated fat cell.
(5) association of cyclic adenosine monophosphate sodium salt, ivy extract, Radix Oenotherae erythrosepalae extract, cocoa extract and L-carnitine
Same-action
In order to confirm cyclic adenosine monophosphate sodium salt, ivy extract, Radix Oenotherae erythrosepalae extract, cocoa extract and L-carnitine
Whether there is synergistic effect when reducing fat, with the pharmaceutical composition of above-mentioned each component to the 3T3- induced by Preparatory work of experiment
L1 fat cell is handled.It is identical when the concentration of every kind of component is with independent processing.As a result as shown in Figure 5, wherein * * P <
0.01, * P < 0.001 * *.
Fig. 5 is the results show that compared with the control group, and fat mass significantly reduces in pharmaceutical composition treated fat cell,
And lipolytic effect is substantially better than the known positive comparative control caffeine with the effect that reduces fat.
(6) cyclic adenosine monophosphate sodium salt, ivy extract, Radix Oenotherae erythrosepalae extract, cocoa extract and L-carnitine combination
The effect that object promotes obese mouse belly fat to decompose
Using aforementioned pharmaceutical compositions as treatment group, the effect of lipolysis is carried out to obesity mice test.In order to examine
The effect that pharmaceutical composition promotes obese mouse belly fat to decompose is tested to press using the obesity mice formed in experimental method 2
The method of experimental method 2 calculates the thickness of mouse weight and abdominal subcutaneous fat.As a result as shown in Figure 6, wherein A: weight, * P <
0.05, * P < 0.01 *；B: subcutaneous fat, P < 0.01 * *.
Fig. 6 is the results show that the pharmaceutical composition can significantly reduce the weight of mouse and the thickness of abdominal subcutaneous fat.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of range is protected, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should
Understand, it can be with modification or equivalent replacement of the technical solution of the present invention are made, without departing from the essence of technical solution of the present invention
And range.
Claims (10)
1. application of the cyclic adenosine monophosphate salt in preparation external application fat-eliminating slimming product, which is characterized in that the cyclic adenosine monophosphate salt
For compound of formula I, wherein X indicates salt ion
2. application as described in claim 1, which is characterized in that the cyclic adenosine monophosphate salt is cyclic adenosine monophosphate and metal ion
The pharmaceutically acceptable salt of formation.
3. application as claimed in claim 2, which is characterized in that the pharmaceutically acceptable salt is sodium salt.
4. application as described in claim 1, which is characterized in that the product is drug, health care product or cosmetics.
5. application as described in claim 1, which is characterized in that the external application fat-eliminating slimming product is to reduce rouge in fat cell
The product of fat amount.
6. a kind of composition of external application fat-eliminating slimming, which is characterized in that the composition contains cyclic adenosine monophosphate salt.
7. composition as claimed in claim 6, which is characterized in that the composition also contains ivy extract, oenothera biennis
At least one of extract, cocoa extract, L-carnitine.
8. a kind of product of external application fat-eliminating slimming, which is characterized in that the product includes composition described in claim 6 or 7.
9. product as claimed in claim 8, which is characterized in that the product is drug, health care product or cosmetics.
10. product as claimed in claim 8, which is characterized in that the dosage form of the product be paste, paste, paint, patch,
Gelling agent, liniment or spray.
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| CN115227706A (en) * | 2022-06-08 | 2022-10-25 | 珍奥集团股份有限公司 | Application of 5' -monophosphate nucleotide composition in preparation of fat-reducing and weight-losing functional food and medicine |
| WO2023236740A1 (en) * | 2022-06-08 | 2023-12-14 | 陈玉松 | Use of 5'-monophosphate nucleotide composition in preparation of fat-reducing and weight-losing functional food and drug |
| CN115227706B (en) * | 2022-06-08 | 2023-12-29 | 陈玉松 | Application of nucleotide 5' -monophosphate composition in preparation of fat-reducing and weight-losing functional foods and medicines |
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Application publication date: 20190329 |