CN102847138A - Medicine composition containing muscular amino acid and nucleoside compound and preparation method thereof - Google Patents
Medicine composition containing muscular amino acid and nucleoside compound and preparation method thereof Download PDFInfo
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- CN102847138A CN102847138A CN2012103578885A CN201210357888A CN102847138A CN 102847138 A CN102847138 A CN 102847138A CN 2012103578885 A CN2012103578885 A CN 2012103578885A CN 201210357888 A CN201210357888 A CN 201210357888A CN 102847138 A CN102847138 A CN 102847138A
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Abstract
The invention provides a medicine composition containing a muscular amino acid and nucleoside compound and a preparation method thereof. The medicine composition comprises the following components by weight proportion: 1-5 of muscular amino acid and nucleoside, 1-5 of acetone chloroform, 1.5-2.5 of sodium dihydrogen phosphate and 500-10000 of injection water, and the pH is 6.0-6.5. The medicine composition needs no high-temperature sterilization during preparation, a 4kDa ultrafilter is adopted during purification of proteins for ultra-filtering, and effective components are preserved well. By adding the sodium dihydrogen phosphate to regulate and stabilize pH and combining the sodium dihydrogen phosphate and acetone chloroform for use, stability of the muscular amino acid and nucleoside is obviously improved.
Description
Technical field
The present invention relates to field of medicaments, be specifically related to a kind of compound recipe sarcosine peptide aglycone pharmaceutical composition, be used for the diseases such as hepatic dysfunction that adjuvant treating hepatitis causes, the invention still further relates to the preparation method of this medicine.
Background technology
Sarcosine peptide aglycone is the multiple vasoactive peptides that is extracted by healthy rabbits muscle and cardiac muscle, its main component comprises atrial natriuretic peptide, nucleotide (nucleoside) classes such as calcitonin-gene-related peptide and adenosine triphosphate (adenosine) contain 20 several amino acids and metabolic intermediate thereof in addition.Nucleotide acid and several amino acids (essential amino acids) are the important substance that participates in the human life activity.Cardiovascular system diseases is improved disturbance of blood circulation, reduces vascular resistance, increases cardiac muscle and utilize the effects such as oxygen, can promote that hemopoietic system is movable to strengthen, white blood cell count increases, and the blood vessel elasticity of increasing is arranged simultaneously, prevent the sclerosis of blood vessels effect.Can be used for amyotrophy, nervous edema, cerebrovascular accident paralysis, nerve asthenia syndrome, apoplexy, the auxiliary treatment of Aging and peripheral nerve disease due to the cerebral blood supply insufficiency.
The many extraction processes to sarcosine peptide aglycone of prior art are studied, for example patent documentation CN101584713A discloses the step of preparation sarcosine peptide aglycone, comprise the clean stripping and slicing of thawing, homogenate, deeply freeze, heated and boiled is separated supernatant, acid-base precipitation, alkali deposited, ultrafiltration, hot repressing, deeply freeze, ultrafiltration, then detect polypeptide and the hypoxanthic ratio of regulating, finally by making preparation after the ultrafiltration, the ultrafilter of use 3KDa filters calcitonin-gene-related peptide (CGRP) (contain 37 amino acid molecular amounts and be about 3800D) is caused damage in its preparation process.CN1939533B also discloses a kind of injection sarcosine peptide aglycone powdery injection and preparation method thereof, and it at the hypoxanthine that extracts wherein, and pays close attention to wherein atrial natriuretic peptide and calcitonin related peptides to the design emphasis of Cor Leporis flesh.
Dosage form for sarcosine peptide aglycone adopts lyophilized powders at present, for example CN1939533B is exactly disclosed a kind of lyophilized injectable powder, CN1611261A also discloses a kind of lyophilized injectable powder, at present commercially available sarcosine peptide aglycone pharmaceutical composition is also mostly is that (CN101584713A discloses injection for medicine type with lyophilized injectable powder, yet this injection is not made any stability test), this mainly is because the polypeptide in the sarcosine peptide glycoside injection liquid, aminoacid, the material such as nucleoside and nucleotide, in storage, transportation, be vulnerable to such environmental effects in the sales process, particularly in summer in south China area, temperature can reach 40 ℃, easily cause unstable product quality, increase the incidence rate (CN1939533B) of untoward reaction.In addition, because sarcosine peptide aglycone belongs to the peptide series products, unsuitable high temperature sterilize after canned, otherwise its active component can be had a strong impact on, but the unsterilised impact that easily is subject to again microorganism, effect duration is short, and easily produces drug safety.The mode of employing lyophilized powder can solve the defective of injection form to a certain extent, yet lyophilized powder needs loaded down with trivial details preparation process, need expensive equipment and higher energy consumption, comprehensive preparation cost is high, and often need to add excipient such as mannitol, dextran, lactose, sorbitol, citric acid, these excipient more or less can produce certain side effect, thereby may affect drug safety.
Summary of the invention
The object of the invention is to for above-mentioned deficiency, a kind of novel sarcosine peptide aglycone pharmaceutical composition is provided.
For achieving the above object, technical scheme of the present invention is as follows:
A kind of sarcosine peptide aglycone compound medicine compositions that contains, it comprises the composition of following weight ratio: sarcosine peptide aglycone 1～5, chlorobutanol 1～5, sodium dihydrogen phosphate 1.5～2.5, water for injection 500～10000, pH6.0～6.5.Preferably include the composition of following weight ratio: sarcosine peptide aglycone 2～4, chlorobutanol 2～4, sodium dihydrogen phosphate 1.8～2.2, water for injection 1000～5000, pH6.2～6.4.The composition that more preferably comprises following weight ratio: sarcosine peptide aglycone 3, chlorobutanol 3, sodium dihydrogen phosphate 2.0, water for injection 1500, pH6.3.
Polypeptide and hypoxanthine weight ratio are 5～10: 1 in the wherein said sarcosine peptide aglycone, are preferably 7: 1.
The present invention also provides the method for preparing aforementioned pharmaceutical compositions, it is characterized in that, the method comprises the steps:
1) pretreatment: Cor Leporis flesh and rabbit muscle are rejected fat and connective tissue, clean, be cut into less than 1cm respectively
3Fritter, again clean;
2) homogenate: pretreated Cor Leporis flesh and rabbit muscle added respectively 2～2.5 times purified water, the homogenate of making respectively 2～3 microns fineness;
3) freeze thawing: with the homogenate of Cor Leporis flesh and rabbit muscle homogenate multigelation 2～4 times, whenever inferior to-40 ℃ of lower dark freezing 4～8 hours, and under 45 ℃, thaw;
4) remove impurity: pH to 3.0～4.0 are regulated respectively in the Cor Leporis flesh homogenate after freeze thawing and rabbit muscle homogenate boiled 10～15 minutes, cooling, layer deoils, in the centrifugal 5～10min of 2500～3500rpm, get supernatant, again in the centrifugal 5～10min of 2500～3500rpm, get supernatant after precipitation is dissolved with water for injection, merge supernatant;
5) ultrafiltration: supernatant is regulated pH6～7, filters through 10,000 daltonian ultrafilters first, filters through 4000 daltonian ultrafilters again;
6) detect adjusting: detect Cor Leporis flesh and extract polypeptide and hypoxanthic amount and rabbit muscle extraction polypeptide and hypoxanthic amount, adjusting mixed proportion, to make polypeptide and hypoxanthine weight ratio be to obtain amido peptide glycosides at 5～10: 1, if hypoxanthic quantity not sufficient also can additionally be added hypoxanthine, complement to described ratio;
7) preparation: chlorobutanol, the sodium dihydrogen phosphate of recipe quantity are dissolved with water for injection, add the amido peptide glycosides after regulating of recipe quantity, mix homogeneously adds to the full amount of water for injection, and regulates pH, and is canned behind the fine straining.
Sarcosine peptide aglycone pharmaceutical composition of the present invention is ejection preparation, it has added chlorobutanol, chlorobutanol has antibacterial antisepsis, therefore and the invention compositions does not need to take the high temperature sterilize step after canned, can avoid the loss of effective ingredient, chlorobutanol also has analgesic activity in addition, can alleviate patient's pain.It has been generally acknowledged that, may there be irritated grade for side effect in chlorobutanol, but does not find anaphylaxis in test of the present invention, shows that its application in the sarcosine peptide aglycone pharmaceutical composition is safe.The present invention also regulates by sodium dihydrogen phosphate and stablizes pH, by stablizing the use in conjunction of pH and chlorobutanol, so that the stability of sarcosine peptide aglycone of the present invention obviously strengthens.In addition, pharmaceutical composition of the present invention uses the ultrafilter ultrafiltration of 4kDa in preparation process, can prevent that effective ingredient is by filtering.
The specific embodiment
Following examples are used for further specifying the present invention, but should not be construed as limitation of the present invention.Under the prerequisite that does not deviate from the present invention's spirit and essence, modification made for the present invention or replacement all belong to category of the present invention.
Embodiment 1
1) pretreatment: Cor Leporis flesh and rabbit muscle are rejected fat and connective tissue, clean, be cut into respectively the fritter less than 1cm3, again clean;
2) homogenate: pretreated Cor Leporis flesh and rabbit muscle added respectively 2 times purified water, the homogenate of making respectively 2～3 microns fineness;
3) freeze thawing: with the homogenate of Cor Leporis flesh and rabbit muscle homogenate multigelation 3 times, whenever inferior to-40 ℃ of lower dark freezing 6 hours, and under 45 ℃, thaw;
4) remove impurity: pH to 3.5 is regulated respectively in the Cor Leporis flesh homogenate after freeze thawing and rabbit muscle homogenate boiled 15 minutes, be cooled to room temperature, layer deoils, in the centrifugal 10min of 3000rpm, get supernatant, precipitation with after the water for injection dissolving again in the centrifugal 10min of 3000rpm, get supernatant, merge supernatant;
5) ultrafiltration: supernatant is regulated pH6.5, filters through 10,000 daltonian ultrafilters first, filters through 4000 daltonian ultrafilters again;
6) detect to regulate: detect Cor Leporis flesh and extract polypeptide and hypoxanthic amount and rabbit muscle and extract polypeptide and hypoxanthic amount, adjusting mixed proportion, to make polypeptide and hypoxanthine weight ratio be to obtain amido peptide glycosides at 7: 1;
7) preparation: chlorobutanol 3g, sodium dihydrogen phosphate 2g are dissolved with water for injection, the amido peptide glycosides (polypeptide and hypoxanthine content 3g) of adding after regulating, mix homogeneously injects water to 1500, and regulate pH to 6.3, canned behind the 0.22 μ m membrane filtration.
Embodiment 2
1) pretreatment: Cor Leporis flesh and rabbit muscle are rejected fat and connective tissue, clean, be cut into respectively the fritter less than 1cm3, again clean;
2) homogenate: pretreated Cor Leporis flesh and rabbit muscle added respectively 2.5 times purified water, the homogenate of making respectively 2～3 microns fineness;
3) freeze thawing: with the homogenate of Cor Leporis flesh and rabbit muscle homogenate multigelation 4 times, whenever inferior to-40 ℃ of lower dark freezing 4 hours, and under 45 ℃, thaw;
4) remove impurity: pH to 3.0 is regulated respectively in the Cor Leporis flesh homogenate after freeze thawing and rabbit muscle homogenate boiled 10 minutes, be cooled to room temperature, layer deoils, in the centrifugal 5min of 3500rpm, get supernatant, precipitation with after the water for injection dissolving again in the centrifugal 5min of 3500rpm, get supernatant, merge supernatant;
5) ultrafiltration: supernatant is regulated pH6.0, filters through 10,000 daltonian ultrafilters first, filters through 4000 daltonian ultrafilters again;
6) detect to regulate: detect Cor Leporis flesh and extract polypeptide and hypoxanthic amount and rabbit muscle and extract polypeptide and hypoxanthic amount, adjusting mixed proportion, to make polypeptide and hypoxanthine weight ratio be to obtain amido peptide glycosides at 5: 1;
7) preparation: chlorobutanol 4g, sodium dihydrogen phosphate 2.2g are dissolved with water for injection, the amido peptide glycosides (polypeptide and hypoxanthine content 5g) of adding after regulating, mix homogeneously injects water to 10000, and regulate pH to 6.0, canned behind the 0.22 μ m membrane filtration.
Embodiment 3
1) pretreatment: Cor Leporis flesh and rabbit muscle are rejected fat and connective tissue, clean, be cut into respectively the fritter less than 1cm3, again clean;
2) homogenate: pretreated Cor Leporis flesh and rabbit muscle added respectively 2.5 times purified water, the homogenate of making respectively 2～3 microns fineness;
3) freeze thawing: with the homogenate of Cor Leporis flesh and rabbit muscle homogenate multigelation 2 times, whenever inferior to-40 ℃ of lower dark freezing 8 hours, and under 45 ℃, thaw;
4) remove impurity: pH to 3.0 is regulated respectively in the Cor Leporis flesh homogenate after freeze thawing and rabbit muscle homogenate boiled 10 minutes, be cooled to room temperature, layer deoils, in the centrifugal 5min of 3500rpm, get supernatant, precipitation with after the water for injection dissolving again in the centrifugal 10min of 2500rpm, get supernatant, merge supernatant;
5) ultrafiltration: supernatant is regulated pH6.0, filters through 10,000 daltonian ultrafilters first, filters through 4000 daltonian ultrafilters again;
6) detect to regulate: detect Cor Leporis flesh and extract polypeptide and hypoxanthic amount and rabbit muscle and extract polypeptide and hypoxanthic amount, adjusting mixed proportion, to make polypeptide and hypoxanthine weight ratio be to obtain amido peptide glycosides at 10: 1;
7) preparation: chlorobutanol 2g, sodium dihydrogen phosphate 1.8g are dissolved with water for injection, the amido peptide glycosides (polypeptide and hypoxanthine content 2g) of adding after regulating, mix homogeneously injects water to 1000, and regulate pH to 6.2, canned behind the 0.22 μ m membrane filtration.
Embodiment 4
The preparation method of the sarcosine peptide glycoside injection liquid in this example is identical with embodiment 1, and filling a prescription is: sarcosine peptide aglycone 1g, chlorobutanol 2, sodium dihydrogen phosphate 1.5g, water for injection 500, pH6.4.
Experimental example 1: safety experiment (sensitization experiment)
Healthy guinea pig, body weight 300 ± 30g is divided into 6 groups at random, and 6 every group, male and female half and half.According to matched group (not adding chlorobutanol and sodium dihydrogen phosphate by embodiment 1 preparation), every other day only gives lumbar injection injection 0.5ml/ to experimental group 1～5 group (embodiment 1～5) respectively, continuous three times, then attacked respectively administration (2ml/ only) in the 14th day and 21 days, observed immediately 1 hour.Result's demonstration, obvious abnormal phenomena does not all appear in matched group and experimental group, and both differences are not remarkable, show that its anaphylaxis is negative.
Experimental example 2: stability experiment
The sarcosine peptide aglycone of embodiment 1～5 preparation is carried out detection and long-term (0,1,3,6 month) stability test of the indexs such as clarity, related substance and content, contrast is for according to embodiment 1 preparation but do not add chlorobutanol and sodium dihydrogen phosphate, and the result is as follows:
Table 1 sample indices testing result
| Sample | Clarity | PH value | Related substance (%) | Content (%) |
| Embodiment 1 | Yellow clear liquid | 6.3 | 1.24 | 100.0 |
| Embodiment 2 | Yellow clear liquid | 6.0 | 1.23 | 99.9 |
| Embodiment 3 | Yellow clear liquid | 6.2 | 1.24 | 99.9 |
| Embodiment 4 | Yellow clear liquid | 6.4 | 1.25 | 99.8 |
| Contrast | Yellow clear liquid | 6.5 | 1.24 | 99.9 |
Described sample is placed indices testing result after 1 month:
Table 2 sample indices testing result (1 month)
| Sample | Clarity | PH value | Related substance (%) | Content (%) |
| Embodiment 1 | Yellow clear liquid | 6.3 | 1.24 | 99.8 |
| Embodiment 2 | Yellow clear liquid | 6.1 | 1.24 | 99.8 |
| Embodiment 3 | Yellow clear liquid | 6.2 | 1.25 | 99.7 |
| Embodiment 4 | Yellow clear liquid | 6.4 | 1.26 | 99.8 |
| Contrast | Yellow clear liquid | 6.7 | 1.29 | 99.2 |
Described sample is placed indices testing result after 3 months:
Table 3 sample indices testing result (placing 3 months)
| Sample | Clarity | PH value | Related substance (%) | Content (%) |
| Embodiment 1 | Yellow clear liquid | 6.4 | 1.25 | 99.6 |
| Embodiment 2 | Yellow clear liquid | 6.2 | 1.25 | 99.5 |
| Embodiment 3 | Yellow clear liquid | 6.3 | 1.26 | 99.5 |
| Embodiment 4 | Yellow clear liquid | 6.5 | 1.26 | 99.6 |
| Contrast | Yellow clear liquid | 6.9 | 1.35 | 98.7 |
Described sample is placed indices testing result after 6 months:
Table 4 sample indices testing result (placing 6 months)
| Sample | Clarity | PH value | Related substance (%) | Content (%) |
| Embodiment 1 | Yellow clear liquid | 6.4 | 1.26 | 99.4 |
| Embodiment 2 | Yellow clear liquid | 6.3 | 1.25 | 99.3 |
| Embodiment 3 | Yellow clear liquid | 6.4 | 1.24 | 99.2 |
| Embodiment 4 | Yellow clear liquid | 6.5 | 1.25 | 99.3 |
| Contrast | Precipitation appears | 7.1 | 1.41 | 97.3 |
Above experimental result can be found out: injection provided by the invention meets the regulation of the countries concerned's standard, places for a long time rear stability high, and the indices such as clarity, pH, related substance and content change all not obvious.
Although above used general explanation, the specific embodiment and experiment, the present invention is described in detail, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (5)
1. one kind contains sarcosine peptide aglycone compound medicine compositions, and it comprises the composition of following weight ratio: sarcosine peptide aglycone 1～5, chlorobutanol 1～5, sodium dihydrogen phosphate 1.5～2.5, water for injection 500～10000, pH6.0～6.5.
2. pharmaceutical composition according to claim 1, it comprises the composition of following weight ratio: sarcosine peptide aglycone 2～4, chlorobutanol 2～4, sodium dihydrogen phosphate 1.8～2.2, water for injection 1000～5000, pH6.2～6.4.
3. pharmaceutical composition according to claim 2, it comprises the composition of following weight ratio: sarcosine peptide aglycone 3, chlorobutanol 3, sodium dihydrogen phosphate 2.0, water for injection 1500, pH6.3.
4. each described pharmaceutical composition is characterized in that according to claim 1～3, and polypeptide and hypoxanthine weight ratio are 5～10: 1 in the described sarcosine peptide aglycone.
5. prepare the method for each described pharmaceutical composition of claim 1～4, it is characterized in that, the method comprises the steps:
1) pretreatment: Cor Leporis flesh and rabbit muscle are rejected fat and connective tissue, clean, be cut into less than 1cm respectively
3Fritter, again clean;
2) homogenate: pretreated Cor Leporis flesh and rabbit muscle added respectively 2～2.5 times purified water, the homogenate of making respectively 2～3 microns fineness;
3) freeze thawing: with the homogenate of Cor Leporis flesh and rabbit muscle homogenate multigelation 2～4 times, whenever inferior to-40 ℃ of lower dark freezing 4～8 hours, and under 45 ℃, thaw;
4) remove impurity: pH to 3.0～4.0 are regulated respectively in the Cor Leporis flesh homogenate after freeze thawing and rabbit muscle homogenate boiled 10～15 minutes, cooling, layer deoils, in the centrifugal 5～10min of 2500～3500rpm, get supernatant, again in the centrifugal 5～10min of 2500～3500rpm, get supernatant after precipitation is dissolved with water for injection, merge supernatant;
5) ultrafiltration: supernatant is regulated pH6～7, filters through 10,000 daltonian ultrafilters first, filters through 4000 daltonian ultrafilters again;
6) detect to regulate: detect Cor Leporis flesh and extract polypeptide and hypoxanthic amount and rabbit muscle and extract polypeptide and hypoxanthic amount, adjusting mixed proportion, to make polypeptide and hypoxanthine weight ratio be to obtain amido peptide glycosides at 5～10: 1;
7) preparation: chlorobutanol, the sodium dihydrogen phosphate of recipe quantity are dissolved with water for injection, add the amido peptide glycosides after regulating of recipe quantity, mix homogeneously adds to the full amount of water for injection, and regulates pH, and is canned behind the fine straining.
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| CN2012103578885A CN102847138A (en) | 2012-09-25 | 2012-09-25 | Medicine composition containing muscular amino acid and nucleoside compound and preparation method thereof |
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| CN2012103578885A CN102847138A (en) | 2012-09-25 | 2012-09-25 | Medicine composition containing muscular amino acid and nucleoside compound and preparation method thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824294A (en) * | 2005-12-21 | 2006-08-30 | 白求恩医科大学制药厂 | Preparation technology of sarcosine peptide glycoside injection liquid |
| CN101584713A (en) * | 2009-07-03 | 2009-11-25 | 石海 | Muscular amino acids and peptides and nucleosides injection and preparing method thereof |
| CN102525899A (en) * | 2012-01-17 | 2012-07-04 | 山东罗欣药业股份有限公司 | Injection solution of oxiracetam composition and preparation method thereof |
-
2012
- 2012-09-25 CN CN2012103578885A patent/CN102847138A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824294A (en) * | 2005-12-21 | 2006-08-30 | 白求恩医科大学制药厂 | Preparation technology of sarcosine peptide glycoside injection liquid |
| CN101584713A (en) * | 2009-07-03 | 2009-11-25 | 石海 | Muscular amino acids and peptides and nucleosides injection and preparing method thereof |
| CN102525899A (en) * | 2012-01-17 | 2012-07-04 | 山东罗欣药业股份有限公司 | Injection solution of oxiracetam composition and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 孙妙囡等: "肌氨肽苷制备工艺及其病毒去除方法的验证", 《中国生化药物杂志》 * |
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Application publication date: 20130102 |