CN104606179A - Application of chlorogenic acid in preparation of medicines for treating osteopetrosis - Google Patents
Application of chlorogenic acid in preparation of medicines for treating osteopetrosis Download PDFInfo
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Abstract
The invention provides an application of chlorogenic acid in preparation of medicines for treating osteopetrosis. The chlorogenic acid can be used for effectively treating the osteopetrosis, the treatment effect is better than that of a combined drug of positive drugs such as prednisone and channels-soothing and blood-activating tablets, and the chlorogenic acid is proved to be a safe drug, has small side effects and is good in clinical application prospects.
Description
Technical field
The present invention relates to the purposes of chlorogenic acid in the medicine of preparation treatment osteopetrosis.
Background technology
Osteopetrosis, also known as Marble bone disease, or claim constitutional gristle sclerosis, be a kind of rare dysplastic congenital diseases of general bone structure, skull is one of predilection site.Sclerotin is very close, and loses original structure, and just as marble, but the increase of bone fragility is easily fractured.Still can accompany the situations such as anemia, ophthalmatrophy and deafness.It is generally acknowledged, namely most humans has started there is pathological changes in utero.Be divided into pernicious (child's type) and optimum (adult type) two kinds according to clinical manifestation, the former is often for dying from anemia, poor prognosis after stillbirth or birth.The pathogenic factor of Osteopetrosis is still not clear, may be abnormal relevant with bone resorption, and cause calcium salt excess deposition in bone, outward appearance is marble or Dens Elephatis sample, and fragility increases.Primary disease has family history, is more common in the children of consanguineous marriage.
At present, effective therapy approach of osteopetrosis only has hematopoietic stem cell transplantation, but finds the transplantation donor of hematopoietic stem cell but very difficult.Based on the difficulty and pathogenetic indefinite for the treatment of, all adopt the therapeutic scheme of administering drug combinations symptomatic treatment under normal circumstances, as treated anemia with prednisone, relax through blood circulation promoting slice treatment bone injury, but therapeutic effect is not good, and toxic and side effects is large, as, prednisone can cause femur head necrosis, stress ulcer, steroidal sex hyperglycemia, easy concurrent infection and psychological problem etc.
Have no the report of chlorogenic acid treatment osteopetrosis.
Summary of the invention
Technical scheme of the present invention there is provided the novelty teabag of chlorogenic acid.
The invention provides the purposes of chlorogenic acid in the medicine of preparation treatment osteopetrosis.
Wherein, described medicine be with the chlorogenic acid of effective dose for active component, add the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
Wherein, in described pharmaceutical preparation, every preparation unit contains chlorogenic acid 1 ~ 1000mg.
Wherein, the using dosage of described pharmaceutical preparation Content of Chlorogenic Acid is 1 ~ 100mg/kg.
Wherein, described medicament is oral formulations or injection.
To sum up, chlorogenic acid of the present invention effectively can treat osteopetrosis, therapeutic effect than positive drug prednisone with relax also good through the drug combination of blood circulation promoting slice, and it has been proved to be a kind of safe drugs, and side effect is little, and potential applicability in clinical practice is good.
Obviously, according to foregoing of the present invention, according to ordinary technical knowledge and the customary means of this area, not departing under the present invention's above-mentioned basic fundamental thought prerequisite, the amendment of other various ways, replacement or change can also be made.
The detailed description of the invention of form by the following examples, is described in further detail foregoing of the present invention again.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example.All technology realized based on foregoing of the present invention all belong to scope of the present invention.
Accompanying drawing explanation
Fig. 1 respectively organizes the expression of CA2 gene in mice osteoclast
Fig. 2 respectively organizes the testing result of II type carbonic anhydrase in mice osteoclast.* p<0.05, * * p<0.01 compared with normal group; #p<0.05, ##p<0.01 compared with model group; △ p<0.05, △ △ p<0.01 compared with administering drug combinations group.
Detailed description of the invention
Embodiment 1: pharmacodynamics test research in chlorogenic acid treatment osteosclerosis sick body
1. experiment material
1.1 animal
Osteopetrosis BALB/C mice 50.
The animal model used in the present embodiment, be directly buy suffer from osteopetrosis BALB/C mice (this mice is also the common model of domestic and international osteopetrosis basic research).This mouse model is due to sick Os Mus matter growth failure, and sclerotin is hard, and medullary cavity disappears, and all skeletons all harden, and with anodontia.The pathological changes of this Mus similar to mankind's os osseum disease.
1.2 Experimental agents and instrument
Chlorogenic acid, prednisone (the prosperous milky way Chemical Co., Ltd. in Hebei), tablet for relieving rigidity of muscles and promoting blood circulation (Henan Zhong Jie Pharmaceutical Co., Ltd), distilled water, 12 hole sterile plate, 96 hole sterile plate, high power microscope, electronic balance (100,000/).
2. experimental technique
2.1 laboratory animal grouping and administrations
Osteopetrosis belongs to rare disease, and the clinical treatment to it and research all compare shortcoming, and still do not treat the specific drug of osteopetrosis both at home and abroad, medication rests in the aspect of symptomatic treatment mostly.This experiment adopts the dosage regimen of prednisone and tablet for relieving rigidity of muscles and promoting blood circulation drug combination as positive controls.By dosage: prednisone (6mg/kg), tablet for relieving rigidity of muscles and promoting blood circulation (1/only) carry out drug treatment.Using common BALB/C mice as Normal group.
Osteopetrosis mouse model stochastic averagina is divided into 5 groups, called after model control group (model group respectively, n=10), prednisone and tablet for relieving rigidity of muscles and promoting blood circulation therapeutic alliance group (therapeutic alliance group, n=10), chlorogenic acid low dose therapy group (L-LYS group, n=10), dosage treatment group (M-LYS group in chlorogenic acid, and chlorogenic acid high-dose therapy group (H-LYS group n=10), n=10), together with Normal group (NC group, n=10), totally 6 experimental grouies, totally 60.Concrete grouping and administering mode as shown in table 1:
Table 1. tests grouping, administering mode and dosage (note: the administration volume of all administration groups is identical)
2.2. Testing index and experimental technique
2.2.1 mouse red blood cell and platelet Indexs measure
Adopt the mode of tail venous blood sampling to get blood to each group of mice, afterwards the erythrocyte in blood and platelet index etc. are detected.
2.2.2 the detection of mice macroscopic features, survival rate regulating liver-QI spleen weight
The macroscopic features, body weight change etc. of each group of mice are detected.After experiment terminates, put to death mice, take liver and the spleen weight of each group of mice, calculate liver, spleen percentage ratio (liver or spleen weight/Mouse Weight).
2.2.3 mice osteocomma absorption lacuna counting and absorption lacuna area estimation
1. the separation of osteoclast
The long bone of limbs of the mice after upper step being dissected is separated, careful removing is attached to soft tissue and the epiphysis of bone surface, diaphyseal portion PBS puts in the glass dish filling 15%199 culture fluid (containing 10% calf serum, 5% hyclone, 100 μ g/mL streptomycin sulfates, 100U/mL penicillin sodium, pH7.2) after cleaning.Gently scraped into culture fluid by sclerotin inner surface with dissecting knife, more repeatedly blow and beat sclerotin fragment 2min by round end custom, after standing 30s, by upper strata cell suspension, (density is 1 × 10
6/ mL) be evenly inoculated in 24 well culture plates of preset thin osteocomma.Every hole adds osteoclast suspension 1mL and the cell culture fluid 1mL containing serum respectively, cultivates.
2. osteocomma absorption lacuna technology and absorption lacuna area estimation
Count the absorption lacuna number on each group of osteocomma under adopting osteocomma absorption lacuna Toluidine blue staining light microscopic method 100 times, carry out recording and analyzing.
3. experimental result
3.1 mouse red blood cells and platelet Indexs measure
The erythrocyte of mice and the result display of platelet Indexs measure, suffers from the model group mice of osteopetrosis, its erythrocyte and platelet count significantly reduce, and it is compared with Normal group, have significant differences (p<0.01).The erythrocyte of the mice of administering drug combinations group and platelet count comparatively model group have compared obvious rising, therebetween significant differences (p<0.01) is had, but compared with Normal group, still there is significant difference (p<0.05).The mice of chlorogenic acid treatment group, its hemocyte and platelet count also have remarkable rising, compared with model group, there is significant differences (p<0.01), and the chlorogenic acid administration group of middle and high dosage, erythrocyte and platelet count can be made to reach normal value, and between Normal group, there is no significant difference (p>0.05).The erythrocyte of various dose chlorogenic acid administration group mice and platelet count, all higher than administering drug combinations group, each group data respectively compared with administering drug combinations group after show significant difference (p<0.05).
Table 2 respectively organizes erythrocyte and the platelet measurement result of mice
The detection of 3.2 mice macroscopic features, body weight, survival rate regulating liver-QI, spleen percentage by weight
Treat after 60 days, observe the various appearance of mice and aspectual character, found that: the dead mouse of (1) model group is serious, 10 have merely hit 7 death, and the mice bone of survival is stiff, and muscle also large area is stiff, and twitches with intermittent.(2) mice of administering drug combinations treatment group, relative death rate is lower, and have 4 dead mouses, and the activity of mice is normal, bone stiffness is more weak, and facial muscle are stiff comparatively serious, and also observes mice and have tic phenomenon.(3), in the mice that chlorogenic acid is treated, its mortality rate obviously reduces, the dead mouse of low dosage chlorogenic acid treatment group 1, and mice activity is normal, and bone also has no obviously stiff, only has the face of two mices to have muscle-bound phenomenon.And the mice of middle dosage chlorogenic acid treatment group has no dead, and movable normal, bone is not stiff, and muscle is not stiff yet.The dead mouse of the chlorogenic acid treatment group of high dose one, its activity is normal, and mice bone has no obviously stiff, and face is normal.
Liver, spleen enlargement, it is one of clinical manifestation of osteopetrosis, liver after dissecting the mice of each group and spleen obtain liver, spleen weight percent ratio index after weighing, result shows, compared with normal group, Mouse Liver, the spleen percentage by weight of model group obviously raise, and point out its liver, splenomegaly degree larger.And the mice of administering drug combinations treatment, its liver, spleen percentage by weight all have obvious decline, and have significant difference (p<0.05) between model group.And each group of mice of chlorogenic acid treatment, the percentage by weight of its liver, spleen, also has and comparatively significantly declines, and there is significant difference (p<0.05) between model group.All measurement results are as shown in table 3.
The detection of table 3 macroscopic features regulating liver-QI, spleen percentage by weight
A:P<0.01b:p<0.05 c:p<0.01d:p<0.05 e:p<0.01f:p<0.05 compared with administering drug combinations group compared with model group compared with Normal group
3.3 mice osteocomma absorption lacuna counting and absorption lacuna area estimations
Osteopetrosis be due to osteoclastic bone resorption serious hindrance at all, the result display of this part experiment: the mice of (1) osteopetrosis model group, compared with the mice of Normal group, its osteocomma absorbs nest and falls into number and absorb nest that to fall into area considerably less, points out its osteoclastic bone resorption generation serious hindrance.(2) mice of administering drug combinations treatment group, its osteocomma absorption lacuna number is without obvious rising, and no difference of science of statistics (p>0.05) between model group.(3) mice of high, medium and low dosage chlorogenic acid administration group, its absorption nest falls into area all obvious rising, the increasing degree of each group is different, respectively compared with model group, all has significant differences (p<0.01).The mice of high, middle dosage chlorogenic acid treatment group, the measurement result of its osteocomma absorption lacuna counting and absorption lacuna area, and between Normal group, there is no significant difference (p>0.05).Measurement result shows, and chlorogenic acid significantly can increase osteocomma absorption lacuna counting and absorption lacuna area, promotes osteoclastic bone resorption.And administering drug combinations group does not show corresponding experimental result.All measurement results are as shown in table 4.
Table 4 osteocomma absorption lacuna counting and absorption lacuna area estimation result
4 statistical procedures
The experimental data of continuous variable form with
represent, comparison two sample t-test between 2 groups of data.Adopt SPSS13.0 statistical software, P<0.05 is for there being statistical significance.
5. conclusion
The present embodiment is using osteosclerotic BALB/C mice as experimental model, in view of the specific drug of still not treating osteopetrosis both at home and abroad, so in conjunction with clinical actual therapeutic scheme, the method for prednisone and tablet for relieving rigidity of muscles and promoting blood circulation administering drug combinations is adopted to be used as positive control treatment osteopetrosis mice.By the appearance sign of each group of mice, liver and spleen weight index, survival rate, erythrocyte, platelet, bone resorption nest falls into the mensuration of number and area, result shows, osteopetrosis mice after chlorogenic acid treatment, its survival state is good, appearance signs etc. are comparatively normal, liver, spleen weight is tending towards normalization, and erythrocyte and platelet have had and have significantly improved, in addition, osteoclastic bone resorption nest falls into number and area has had significant raising, high, in, the measurement result of low dosage chlorogenic acid treatment group is all higher than prednisone and tablet for relieving rigidity of muscles and promoting blood circulation administering drug combinations treatment group, prompting chlorogenic acid effectively can promote that the bone of osteoclast absorbs, remarkable erythrocyte and the number of platelets improving mice, the survival state etc. of osteopetrosis is all significantly improved, show good drug effect.
Embodiment 2: chlorogenic acid is on the impact of II type carbonic anhydrase expression of the CA2 gene in osteopetrosis mouse model osteoclast and correspondence thereof.
1. experiment material
1.1 animal
Osteopetrosis BALB/C mice 30.
The animal model used in the present embodiment, be directly buy suffer from osteopetrosis BALB/C mice.This mouse model is due to sick Os Mus matter growth failure, and sclerotin is hard, and medullary cavity disappears, and all skeletons all harden, and with anodontia.The pathological changes of this Mus similar to mankind's os osseum disease.
1.2 Experimental agents and instrument
Chlorogenic acid, prednisone (the prosperous milky way Chemical Co., Ltd. in Hebei), tablet for relieving rigidity of muscles and promoting blood circulation (Henan Zhong Jie Pharmaceutical Co., Ltd), II type carbonic anhydrase ELISA detection kit, PCR instrument, RNA extracts test kit, cDNA chain synthetic agent box, distilled water, 12 holes and 96 orifice plates, microplate reader, electronic balance.
2. experimental technique
2.1 laboratory animal grouping and administrations
Osteopetrosis belongs to rare disease, and the clinical treatment to it and research all compare shortcoming, and still do not treat the specific drug of osteopetrosis both at home and abroad, medication rests in the aspect of symptomatic treatment mostly.This experiment adopts the mode of prednisone and tablet for relieving rigidity of muscles and promoting blood circulation drug combination as positive controls.By dosage: prednisone (6mg/kg), tablet for relieving rigidity of muscles and promoting blood circulation (1/only) carry out drug treatment.Using common BALB/C mice as Normal group.
Osteopetrosis mouse model stochastic averagina is divided into 3 groups, called after model control group (model group respectively, n=10), prednisone and tablet for relieving rigidity of muscles and promoting blood circulation therapeutic alliance group (therapeutic alliance group, and chlorogenic acid treatment group (LYS group n=10), n=10), together with Normal group (n=10), totally 4 experimental grouies, totally 40.Concrete grouping and administering mode as shown in table 5:
Table 5. tests grouping, administering mode and dosage (note: the administration volume of all administration groups is identical)
2.2 test experience
2.2.1RT-PCR method detects the expression of the CA2 gene in each group of Mouse Bone cell.
(1) extraction of the total mRNA of osteoclast
1. the separation of osteoclast: after experiment stops, by mice through 75% alcohol-pickled sterilization 5min, long bone of limbs is separated after drawing neck to put to death, carefully clear soft tissue and the epiphysis being attached to bone surface, diaphyseal portion PBS puts in the glass dish filling 15%199 culture fluid (containing 10% calf serum, 5% hyclone, 100 μ g/mL streptomycin sulfates, 100U/mL penicillin sodium, pH7.2) after cleaning.Gently scraped into culture fluid by sclerotin inner surface with dissecting knife, more repeatedly beat sclerotin fragment 2min by round end custom, collect the cell suspension on upper strata after leaving standstill 30s, all experimental grouies are divided into two parts of A and B, for subsequent use.
2. undertaken centrifugal by above-mentioned cell suspension A, collect osteoclast, numbering name respectively, adds the lysate RL being added with beta-mercaptoethanol in advance of 350 μ l in every part of broken bone, leaves standstill 5min.
3. by aforesaid liquid centrifugal 3min under the condition of 12,000rpm/min, careful Aspirate supernatant uses;
4. in supernatant, slowly add the dehydrated alcohol of 0.5 times of volume, repeatedly blow and beat, until be transferred in adsorption column CR3 after mix homogeneously, the centrifugal 2min of 12,000rpm, discards waste liquid, stays adsorption column;
5. add protein liquid removal RW1 in adsorption column, remove albumen, centrifugal 1min, discards waste liquid afterwards;
6. in adsorption column CR3, DNase I working solution is added, degraded DNA wherein;
7.. after cleaning adsorption column respectively with protein liquid removal and rinsing liquid, adsorption column is put in collecting pipe, opens and ventilate, fully volatilize the residual liquid on pillar;
8. in adsorption column, add 40 μ lRNase free ddH2O, after placing 2min, the centrifugal 2min of 12,000rpm, obtains mRNA sample;
(2) reverse transcription of mRNA
The first chain cDNA that the RNA solution synthesis utilizing cDNA first chain synthetic agent box and said extracted to obtain is corresponding.Concrete process of reverse-transcription operates to specifications, is summarized as follows:
1. get being placed on ice bath without enzyme centrifuge tube of 200 μ l, and in wherein adding following solutions: RNA template 5mL, Oligo (dT) 2Ml, Super Pure dNTP2Ml, RNase-Free ddH2O5.5mL.
2. after centrifugal, the centrifuge tube that aforesaid liquid is housed is positioned in PCR instrument, 70 DEG C hatch 5min after, after brief collection liquid, following reagent is added: 5 × firststrand buffer (containing DTT) 4mL after being transferred to cooled on ice 2min rapidly, RNasin0.5mL, TIANScriptM-MLV (200U) 1mL.
3. centrifuge tube mixed gently and after brief centrifugation, PCR instrument 42 DEG C be set and hatch 50min, afterwards 95 DEG C of heating 5min.20ul RNase-Free ddH is added in the most backward cDNA solution obtained
2o is diluted to 40ul, obtains the first chain cDNA.
4. the product obtained carefully is numbered under being placed on-20 DEG C of conditions and preserve.
(3) RT-PCR is quantitative
EvaGreen fluorescent dye is combined with the DNA of double-strand can produce very strong fluorescence, and by detecting final fluorescence intensity, we can obtain reacting the total amount generating DNA.Carry out RT-PCR to add in fluorescent dye, (1) (2) step synthesized cDNA product, CA2 primer mix homogeneously in test tube after to detect body series and use RNase-free water to replace cDNA products group to contrast for NTC.Each sample all sets β-actin primer sets as relative value, carries out relative quantification.
2.2.2 the mensuration of II type carbonic anhydrase in mice osteoclast is respectively organized
Get the B group osteoclast suspension described in 2.2.1 (1), after piping and druming evenly, careful hatches in 12 orifice plates, tests after 24.After carrying out protein quantification by BCA method, utilize II type carbonic anhydrase ELISA enzyme linked immunological kit to carry out follow-up test, measure II type carbonic anhydrase activity.
3. experimental result
The expression of the CA2 gene in 3.1 each group Mouse Bone cells
The testing result display of CA2 gene, in the osteoclast of normal group mice, the expression of CA2 is relatively high, and in the mice of osteopetrosis model group, the CA2 gene expression in its osteoclast is significantly lowered.After respectively with administering drug combinations and chlorogenic acid treatment, in administering drug combinations group, CA2 gene in mice osteoclast does not significantly rise, it does not have significant difference (p>0.05) compared with model group, and the mice of chlorogenic acid treatment, the expression of CA2 gene in mice osteoclast can be improved significantly, it is compared with model group and administering drug combinations group, all there is the difference (* * p<0.01, ##p<0.01) of very significant.The experimental result of gene test as shown in Figure 1.
The measurement result of II type carbonic anhydrase in 3.2 each group mice osteoclasts
The testing result display of II type carbonic anhydrase, in the osteoclast of osteopetrosis model mice, the content of II type carbonic anhydrase is compared to Normal group, remarkable decline, and the key that II type carbonic anhydrase is osteoclastic bone resorption promotes enzyme, this kind of decline is presented at the bone resorption ability wretched insufficiency of osteoclast in the mouse model of osteopetrosis.Its II type carbonic anhydrase content, compared with matched group, has the difference (* * p<0.01) of very significant.After administering drug combinations treatment, II type carbonic anhydrase content in mice osteoclast does not have significant change, does not have statistical difference (p>0.05) between itself and model group.And in the mice of chlorogenic acid treatment, II type carbonic anhydrase content in its osteoclast has had the rise of highly significant, and there is between administering drug combinations group the difference (△ △ p<0.01) of very significant, but compared with Normal group, its value still has significant difference (* p<0.05).
Result shows, and chlorogenic acid can promote the expression of II type carbonic anhydrase efficiently, thus promotes the bone resorption of osteoclasts.Concrete experimental result as shown in Figure 2.
4. statistical procedures
The experimental data of continuous variable form with
represent, comparison two sample t-test between 2 groups of data.Adopt SPSS13.0 statistical software, P<0.05 is for there being statistical significance.
5. conclusion
In the present embodiment, adopt the BALB/C mice suffering from osteopetrosis as animal pattern, give prednisone and tablet for relieving rigidity of muscles and promoting blood circulation as positive treatment control group to combine.Determine the expression of II type carbonic anhydrase of CA2 gene in each group of mice osteoclast and correspondence respectively.Result shows, and compared with administering drug combinations group, chlorogenic acid effectively can raise the expression of CA2 gene in mice osteoclast, promotes that the content of its corresponding II type carbonic anhydrase promotes simultaneously.II type carbonic anhydrase major function promotes that the bone of osteoclast absorbs, and can improve the osteosclerosis caused because of osteoclast malabsorption significantly to its activation.To sum up, chlorogenic acid can activate II type carbonic anhydrase activity significantly, has certain therapeutical effect to osteopetrosis.
Embodiment 3: prepare lyophilized injectable powder with chlorogenic acid
1. the extraction of chlorogenic acid:
Chlorogenic acid crude drug used in the present embodiment, be obtained by extraction, purification in Folium Eucommiae, purity is 99.38%.
2. the preparation of chlorogenic acid lyophilized injectable powder
2.1 prescriptions:
| Purity is the chlorogenic acid (principal agent) of 99.38% | 40g |
| Mannitol (proppant) | 55g |
| Sodium sulfite (antioxidant) | 4.5g |
Above prescription is dissolved in water for injection completely, after filtration, then uses the degerming microporous filter membrane fine straining of 0.22 μm, after regulating pH, make 2ml injectable powder 1000 altogether according to the routine operation of sterile powder injection, often prop up containing chlorogenic acid 40mg.
Embodiment 4: prepare pill with chlorogenic acid
1. the extraction of chlorogenic acid
The chlorogenic acid used in the present embodiment, be obtained by extraction, purification in Folium Eucommiae, purity is 99.21%.
2. the preparation of chlorogenic acid pill
2.1 prescription
| Purity is the chlorogenic acid (principal agent) of 99.21% | 1g |
| Starch (diluent) | 50g |
| Dextrine powder (binding agent) | In right amount |
| Dehydrated alcohol (wetting agent) | In right amount |
2.2. method for making:
Get appropriate PVP K30, solution is mixed with dehydrated alcohol, get chlorogenic acid and the starch of recipe quantity again, after adopting equivalent dilution method mix homogeneously, add in the alcoholic solution of dextrine powder, abundant stirring is obtained soft material afterwards, and adopt stranding ball legal system to obtain chlorogenic acid pill 1000, every pill is containing chlorogenic acid 1mg.
Embodiment 5: prepare oral solution with chlorogenic acid
1. the extraction of chlorogenic acid
The chlorogenic acid used in the present embodiment, be obtained by extraction, purification in Herba Arctii leaf, purity is 99.29%.
2. the preparation of chlorogenic acid oral solution
2.1 prescription
| Purity is the chlorogenic acid (principal agent) of 99.29% | 200g |
| Sodium sulfite (antioxidant) | 10g |
| Water for injection (solvent) | 100L |
2.2 method for making
Get chlorogenic acid and the sodium sulfite of recipe quantity, be dissolved in 5L water for injection, according to the conventional fabrication process of oral liquid, after filtration, sterile filling becomes 1000 oral liquids, and often propping up oral liquid is 100mL, containing chlorogenic acid 200mg.
Embodiment 6: prepare tablet with chlorogenic acid
1. the extraction of chlorogenic acid:
The chlorogenic acid used in the present embodiment, be obtained by extraction, purification in Flos Lonicerae, purity is 98.57%.
2. the preparation of chlorogenic acid tablet
2.1 prescriptions:
2.2 method for makings:
The present embodiment adopts wet granular compression produces chlorogenic acid tablet processed.(1) measure hypromellose by prescription and make aqueous solution; (2), after getting the chlorogenic acid of recipe quantity, Icing Sugar and lactose mix homogeneously, add hypromellose aqueous solution, after stirring, make soft material; (3) rule of operation of soft material wet granulation routinely will prepared, sieves, obtains uniform granule after dry and granulate; (4) tabletting after being mixed homogeneously with magnesium stearate by obtained granule, makes 1000 tablets altogether, and every sheet is containing chlorogenic acid 100mg.
Embodiment 7: prepare capsule with chlorogenic acid
1. the extraction of chlorogenic acid:
The chlorogenic acid used in the present embodiment, be obtained by extraction, purification in Folium Eucommiae, purity is 99.25%.
2. the preparation of chlorogenic acid capsule:
2.1 prescriptions:
| Purity is the chlorogenic acid of 99.25% | 100g |
| Icing Sugar | 200g |
2.2 method for makings:
Get chlorogenic acid and the starch of recipe quantity, mix homogeneously, add 80% alcoholic solution and make soft material, dry, prepare 2000 capsules according to the conventional fabrication process of capsule after granulate, every capsules is containing chlorogenic acid 50mg.
Embodiment 8: prepare granule with chlorogenic acid
1. the extraction of chlorogenic acid
The chlorogenic acid used in the present embodiment, be obtained by extraction, purification in Folium Eucommiae, purity is 98.92%.
2. the preparation of chlorogenic acid granule
2.1 prescriptions:
| Purity is the chlorogenic acid (principal agent) of 98.92% | 200g |
| Mannitol (diluent) | 100g |
| Sucrose (diluent) | 400g |
| PVP K30 (binding agent) | In right amount |
2.2 method for makings:
Get PVP K30, add water for injection, make solution.After getting the chlorogenic acid of recipe quantity, mannitol and lactose mix homogeneously, add PVP K30 solution, make soft material.According to the conventional fabrication process of granule, soft material is sieved, after dry and granulate, obtains granule.Aseptically subpackage granule, prepares 400 bags of granules, and every bag of granule is containing chlorogenic acid 500mg.
Embodiment 9: prepare powder with chlorogenic acid
1. the extraction of chlorogenic acid:
The chlorogenic acid crude drug that the present embodiment is used, be obtained by extraction, purification in Herba Arctii leaf, purity is 98.82%.
2. the preparation of chlorogenic acid powder:
2.1 prescription
Purity is the chlorogenic acid 1000g of 98.82%
2.2 method for making
Get after recipe quantity chlorogenic acid sieves, according to the conventional fabrication process of powder, aseptic subpackaged one-tenth is containing 1000 bottle/bag powders, and every bottle/bag powder is containing chlorogenic acid 1000mg.
To sum up, chlorogenic acid of the present invention effectively can treat osteopetrosis, and therapeutic effect is obviously better than current normally used osteosis medicine: prednisone and the drug combination relaxed through blood circulation promoting slice, side effect is little, and potential applicability in clinical practice is good.
Claims (5)
1. the purposes of chlorogenic acid in the medicine of preparation treatment osteopetrosis.
2. purposes according to claim 1, is characterized in that: described medicine be with the chlorogenic acid of effective dose for active component, add the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
3. purposes according to claim 2, is characterized in that: in described pharmaceutical preparation, every preparation unit contains chlorogenic acid 1 ~ 1000mg.
4. purposes according to claim 3, is characterized in that: the using dosage of described pharmaceutical preparation Content of Chlorogenic Acid is 1 ~ 100mg/kg.
5. the purposes according to claim 3 or 4, is characterized in that: described medicament is oral formulations or injection.
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