CN105434341B - A kind of polyadenylic-polyuridylic acid parenteral solution for animals and preparation method thereof - Google Patents
A kind of polyadenylic-polyuridylic acid parenteral solution for animals and preparation method thereof Download PDFInfo
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- A61K47/20—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
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Abstract
The invention belongs to pharmaceutical technology fields, and in particular to a kind of polyadenylic-polyuridylic acid parenteral solution for animals and preparation method thereof.Present invention polyadenylic-polyuridylic acid parenteral solution for animals includes compound sodium chloride injection, also comprises the following components and its concentration：4 10g/L polyadenylic-polyuridylic acids, 0.5 4g/L poly-D-lysines, 0.5 3g/L lauryl sodium sulfate.The present invention also provides the preparation methods of the polyadenylic-polyuridylic acid parenteral solution for animals.The polyadenylic-polyuridylic acid for animals injection formula of liquid of the present invention is simple, and safe, it is easy to prepare, suitable for large-scale production, stable quality.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to a kind of polyadenylic-polyuridylic acid parenteral solution for animals and preparation method thereof.
Background technology
Polyadenylic-polyuridylic acid, alias：Polyinosinic acid cytidine, polyinosinic acid, poly IC, PolyI:C
(polynosinic acid-polycyttdylic acid) is a kind of efficient interferon inducer.Polyadenylic-polyuridylic acid be by
Artificial synthesized polyinosinic acid (polyinosinic acid, Poly I) and poly- cytimidine acid (polycytidylic acid,
Poly C) pairing after double stranded RNA, have broad-spectrum disease resistance toxic action and immunosuppressive action, to herpes zoster, bleb
Property keratitis, chronic viral hepatitis, verruca vulgaris, flat wart etc. there is the effect of certain.
Polyadenylic-polyuridylic acid is mainly used in prevention and treatment poultry, virosis of livestock in terms of veterinary drug, has and uses
The characteristics of convenient, extensive, it can simultaneously use with many drugs, with astragalus polyose, levamisol, Ribavirin compatibility, have bright
Aobvious synergistic effect；This product is added in many kinds such as powder, powder, has the function of to significantly improve existing product curative effect, together
When do not interfere the detection of original product.In addition, this product, without species specificity, antiviral spectrum is wide, and small toxicity, safe range are big, not shadow
Body antibody level is rung, avoids that the antibody level caused by booster immunization is excessively high and vaccine stress reaction, can be in extensively
It is used on the animals such as fowl, pig, ox.Polyadenylic-polyuridylic acid uses simultaneously with vaccine, can play the Blank immunization phase that makes up, alleviation vaccine is answered
Swash reaction, the effect of enhancing immune effect.
Polyadenylic-polyuridylic acid is double stranded RNA, is easily degraded by RNA enzyme, generally requires to add in the preparation process of preparation
Add appropriate stabilizer, inhibit RNA enzyme, increase the stability of polyadenylic-polyuridylic acid RNA duplex structures.Current Polyinosinic injection
Contain kanamycins in product as cationic stabilized agent, and the use of kanamycins be easy to cause residual of antibiotic etc. and asks
Topic.In addition, Ca2+Also there is important role to the secondary structure for stablizing poly IC, the prior art is in preparation process directly anti-
Addition calcium chloride powder in liquid is answered, since calcium chloride compares indissoluble in itself, high concentration Ca can be led to by directly adding powder2+Easily with
The ingredients such as the hydrogen phosphate in reaction system form precipitation and influence product quality and synthesis concentration.
Chinese patent application 201410654368.4 discloses Polyinosinic injection pharmaceutical composition and preparation method.The poly IC
Injection formula is complicated, and containing polyinosinic acid, poly, acid-base buffer, osmotic pressure regulator, sorbic acid or its is medicinal
The Multiple components such as salt, polyhydroxy-alcohol, stabilizer and water for injection, and preparation process is complicated.
Invention content
The purpose of the present invention is to provide a kind of ingredient is simple, it is easy to prepare, the polyadenylic-polyuridylic acid for animals note of stable quality
Liquid is penetrated, the present invention also provides the preparation methods of the polyadenylic-polyuridylic acid parenteral solution for animals.
To solve the above problems, the technical solution adopted in the present invention is as follows：
A kind of polyadenylic-polyuridylic acid parenteral solution for animals, which is characterized in that the parenteral solution includes compound sodium chloride injection, also
Including following component and its concentration：4-10g/L polyadenylic-polyuridylic acids, 0.5-4g/L poly-D-lysines, 0.5-3g/L dodecyl sulphur
Sour sodium.
Further, it to match is 1 that the polyadenylic-polyuridylic acid, which is polyinosinic acid and poly- cytimidine acid,:1 is prepared.
Preferably, the polyadenylic-polyuridylic acid parenteral solution for animals include compound sodium chloride injection, also comprise the following components and
Its concentration：8g/L polyadenylic-polyuridylic acids, 4g/L poly-D-lysines, 3g/L lauryl sodium sulfate.
Correspondingly, the present invention provides the preparation method of compound sodium chloride injection, step is：33 mg of calcium chloride is taken,
The water for injection of 100mL is added in, is placed in 60-70 DEG C of water-bath, is stirred continuously to being completely dissolved, adds in 850 mg of sodium chloride and chlorine
Change 30 mg of potassium, continue stirring to solution become clarification to get.
Correspondingly, the present invention provides the preparation method of the polyadenylic-polyuridylic acid parenteral solution for animals, step is：
S1, take polyinosinic acid, add in the ringer's solution of 1/2 amount, be placed in 60-70 DEG C of water-bath, be stirred continuously to
It is completely dissolved, it is spare；
S2, poly- cytimidine acid is taken, adds in the ringer's solution of 1/2 amount, be placed in 60-70 DEG C of water-bath, be stirred continuously
It is spare to being completely dissolved；
S3, the liquid for obtaining S1 and S2 mix, and 10-20min are stirred with the speed of 30-40 r/min, at 60-70 DEG C
At a temperature of react 40-60min, the speed stirring of 30-40 r/min is kept in reaction process, obtains reaction solution；
S4, poly-D-lysine and lauryl sodium sulfate are added in into the reaction solution described in S3, continues to stir to completely molten
Solution, treats that solution temperature is down to room temperature, with 0.20-0.45 μm of filter filtration sterilization, be dispensed into the ampoule bottle of 5mL to get.
Compared with prior art, the invention has the advantages that：The formula letter of present invention polyadenylic-polyuridylic acid parenteral solution for animals
Singly, the quality examinations such as finished product pH value obtained, osmotic pressure, clarity, visible foreign matters inspection, sterility test meet the requirements.This
Polyadenylic-polyuridylic acid parenteral solution for animals is invented using poly-D-lysine and lauryl sodium sulfate as stabilizer, RNA enzyme can be inhibited
Activity increases the stability of polyadenylic-polyuridylic acid RNA duplex structures.Poly-D-lysine has strong bacteriostasis, water-soluble strong,
Stability is good, safe, in vivo the degradable essential amino acid in L-lysine this human body, and poly in addition relies ammonia
Polycation in acid increases the stability of polyadenylic-polyuridylic acid RNA duplex structures.Lauryl sodium sulfate is a kind of nontoxic
Anion surfactant, it is water-soluble strong, it is inhibited to RNA enzyme.In addition the present invention is used in ringer's solution
Contain the Ca in calcium chloride2+Also there are important role, and inventive formulation each component water to the secondary structure for stablizing poly IC
Dissolubility is preferable, can directly with ringer's solution compatibility.Finally, the preparation side of present invention polyadenylic-polyuridylic acid parenteral solution for animals
Method is simple, stable quality, is suitble to large-scale production.
Specific embodiment
It will be understood by those skilled in the art that technology disclosed in following embodiment represent the inventors discovered that in this hair
The technology of good action is played in bright practice.However, many changes can be made in disclosed specific embodiment, and
Still it obtains the same or similar as a result, without departing from the spirit and scope of the present invention.
Embodiment 1
Present invention polyadenylic-polyuridylic acid parenteral solution for animals includes compound sodium chloride injection, also comprises the following components and its dense
Degree：8g/L polyadenylic-polyuridylic acids, 4g/L poly-D-lysines, 3g/L lauryl sodium sulfate.
Preparation method is as follows：
S1, polyinosinic acid is taken, adds in the ringer's solution of 1/2 amount, be placed in 60 DEG C of water-baths, be stirred continuously to complete
Dissolving, it is spare；
S2, poly- cytimidine acid is taken, adds in the ringer's solution of 1/2 amount, be placed in 60 DEG C of water-baths, be stirred continuously to complete
Fully dissolved, it is spare；
S3, the liquid for obtaining S1 and S2 mix, and 15min is stirred with the speed of 40 r/min, anti-at a temperature of 60 DEG C
60min is answered, the speed stirring of 40r/min is kept in reaction process, obtains reaction solution；
S4, poly-D-lysine and lauryl sodium sulfate are added in into the reaction solution described in S3, continues to stir to completely molten
Solution, treats that solution temperature is down to room temperature, with 0.22 μm of filter filtration sterilization, be dispensed into the ampoule bottle of 5mL to get.
Embodiment 2
Present invention polyadenylic-polyuridylic acid parenteral solution for animals includes compound sodium chloride injection, also comprises the following components and its dense
Degree：6g/L polyadenylic-polyuridylic acids, 4g/L poly-D-lysines, 2g/L lauryl sodium sulfate.
The preparation method is the same as that of Example 1.
Embodiment 3
Present invention polyadenylic-polyuridylic acid parenteral solution for animals includes compound sodium chloride injection, also comprises the following components and its dense
Degree：5g/L polyadenylic-polyuridylic acids, 2g/L poly-D-lysines, 2g/L lauryl sodium sulfate.
The preparation method is the same as that of Example 1.
Embodiment 4
Present invention polyadenylic-polyuridylic acid parenteral solution for animals includes compound sodium chloride injection, also comprises the following components and its dense
Degree：10g/L polyadenylic-polyuridylic acids, 4g/L poly-D-lysines, 3g/L lauryl sodium sulfate.
The preparation method is the same as that of Example 1.
Test example one：Quality examination
According to National Drug Administration's national drug standards [WS1-XG-050-2000] to 1-4 of the embodiment of the present invention
Polyadenylic-polyuridylic acid parenteral solution for animals obtained carries out pH value, osmotic pressure, clarity, visible foreign matters inspection, particulate matter, sterile
The quality examinations such as inspection, the results are shown in Table 1.
1 polyadenylic-polyuridylic acid parenteral solution quality examination result for animals of table
As shown in Table 1, polyadenylic-polyuridylic acid parenteral solution for animals made from 1-4 of the embodiment of the present invention meets national drug supervision pipe
The reason office national drug standards [WS1-XG-050-2000], it is quality controllable.
Test example two：Hyperchromicity measures
Polyadenylic-polyuridylic acid for animals made from example 1-4 of the present invention is noted using Fluorescence Increasing experiment and ultra-violet absorption spectrum experiment
Liquid is penetrated to be differentiated, and with commercially available Polyinosinic injection（Chinese medicines quasi-word H2000374, Guangdong Nanguo Medicine Co., Ltd）It carries out
Compare.Wherein Fluorescence Increasing test method is：Polyadenylic-polyuridylic acid parenteral solution 1mL is taken, adds in phenanthrene bromine red solution（Take 0.02mg luxuriant and rich with fragrance
Pyridine bromine it is red add in 100 mL water, dissolve to get）0.5mL is placed in ultraviolet lamp（254nm）Lower observation.Ultra-violet absorption spectrum experiment side
Method is：Polyadenylic-polyuridylic acid parenteral solution 1mL is taken, adds in sodium chloride-phosphate buffer（pH 7.2）, it is made in every mL and about contains
The solution of 0.04mg is measured according to the ultraviolet spectrophotometry of Republic of China Veterinary Pharmacopoeia introduction, the results are shown in Table 2.
2 polyadenylic-polyuridylic acid parenteral solution hyperchromicity measurement result for animals of table
As shown in Table 2, polyadenylic-polyuridylic acid parenteral solution for animals made from 1-4 of the embodiment of the present invention has apparent Fluorescence Increasing
Effect, characteristic spectrum scanning result show that present invention polyadenylic-polyuridylic acid parenteral solution for animals has absorption maximum at 266nm,
There is minimal absorption at 231nm, conform to quality requirements.In addition, the polyadenylic-polyuridylic acid parenteral solution for animals of 1-4 of the embodiment of the present invention is hyperchromic
Effect is apparently higher than commercial product, and better with the product of embodiment 1.
Test example three：Sedimentation coefficient measures
Sedimentation coefficient, reference substance are measured according to the polyacrylamide gel electrophoresis of Republic of China Veterinary Pharmacopoeia introduction
The yeast rna for being 4S for known sedimentation coefficient（tRNA）（Electrophoresis is pure）, measure relative mobility, replication 6 times, and
With commercially available Polyinosinic injection（Chinese medicines quasi-word H2000374, Guangdong Nanguo Medicine Co., Ltd）It is compared.It the results are shown in Table 3.
3 relative mobility measurement result of table
Note：Compared with tRNA,*** P＜ 0.001；Compared with commercially available Polyinosinic injection,### P＜ 0.001.
As shown in Table 3, the mobility of polyadenylic-polyuridylic acid parenteral solution for animals made from 1-4 of the embodiment of the present invention and commercial product
Pole is significantly less than the mobility of standard items tRNA（P＜ 0.001）, illustrate that its sedimentation coefficient is all higher than 4S；In addition, the present invention is real
Product mobility pole made from a 1-4 is applied significantly less than the mobility of commercially available Polyinosinic injection（P＜ 0.001）, illustrate it
Quality is better than commercial product.
Test example four：Inhibit the degradation of RNA enzyme using determined by ultraviolet spectrophotometry product of the present invention
Polyadenylic-polyuridylic acid parenteral solution for animals made from 1-4 of the embodiment of the present invention is taken to be diluted to effective concentration respectively be
0.08mg/mL adds in the RNA enzyme of 20uL a concentration of 10 mg/mL, is placed in 37 DEG C of water-baths, every 15min, take be placed in right amount it is ultraviolet
Spectrophotometer measures absorbance value at 248nm, and with commercially available Polyinosinic injection（Chinese medicines quasi-word H2000374, Guang Dongnan
Pharmaceutcal corporation, Ltd of state）It is compared.It the results are shown in Table 4.
4 polyadenylic-polyuridylic acid parenteral solution for animals of table inhibits RNA enzyme degradation results
As shown in Table 4, polyadenylic-polyuridylic acid parenteral solution for animals made from 1-4 of the embodiment of the present invention and commercial product are in 0-
In the period of 90min, with the extension of time, RNA double-strands are degraded by RNA enzyme, UV absorption enhancing, wherein commercial product exists
The UV absorption intensity of 90min is stronger than product UV absorption intensity made from embodiment 1-4, shows commercial product RNA enzyme quilt
The degree of inhibition not as good as embodiment 1-4, the above result shows that, product can preferably inhibit RNA made from 1-4 of the embodiment of the present invention
Enzymatic activity.
Test example five：Interferon-induced experiment
It takes 48 SPF grades of chickens, 25 ages in days, is randomly divided into 6 groups, every group 8, takes 4 groups to inject the present embodiment 1-4 systems respectively
The polyadenylic-polyuridylic acid parenteral solution for animals obtained, separately takes one group of commercially available Polyinosinic injection of injection（Chinese medicines quasi-word H2000374, Guang Dongnan
Pharmaceutcal corporation, Ltd of state）, remaining one group of injection ringer's solution is as blank control group.Every chicken presses 0.01mg/kg bodies
Re-injection is penetrated, and interval is injected once for 3 days again, and the 12nd, 24 and 36 hour after last time is injected is taken a blood sample, and is surveyed with ELISA method
Determine the alpha interferon content in blood.It the results are shown in Table 5.
5 interferon-induced result of the test of table
Note：Compared with blank control group,*** P＜ 0.001；Compared with commercially available Polyinosinic injection,## P＜ 0.01,### P＜
0.001。
As shown in Table 5, compared with blank control group, polyadenylic-polyuridylic acid for animals made from injection embodiment of the present invention 1-4 is noted
After penetrating liquid and commercial product, the alpha interferon content in chicken body is dramatically increased in 12,24 and 36h poles（P＜ 0.001）, but with
The extension of time, the alpha interferon content in chicken body gradually decrease.In 12,24 and 36h, injection 1-4 of the embodiment of the present invention is made
Polyadenylic-polyuridylic acid parenteral solution for animals after chicken body in alpha interferon content with injection commercial product after chicken body in α interfere
Cellulose content compares, and is respectively provided with pole significant difference（P＜ 0.01 orP＜ 0.001）, show product made from 1-4 of the embodiment of the present invention
Induce the effect for generating alpha interferon in chicken body better than commercial product.
Test example five：Safety testing
It takes 48 SPF grades of chickens, 25 ages in days, is randomly divided into 6 groups, every group 8, injects respectively made from the present embodiment 1-4
Polyadenylic-polyuridylic acid parenteral solution for animals, commercially available Polyinosinic injection（Chinese medicines quasi-word H2000374, Guangdong Nanguo Medicine Co., Ltd）
And ringer's solution.Every chicken is injected by 0.1mg/kg weight, is used in conjunction 3 days, observe after injection animal whether occur it is different
Paradoxical reaction, cuing open for the 7th day after final injection kill all animals, check injection site and visceral organ tissue.
The result shows that：The non-abnormal response of each group animal, off-test after injection, cut open and kill all animals, injection site and
Visceral organ tissue is normal, and it is safe to chicken to illustrate this preparation.
It is any in the spirit and/or range of the present invention due to describing the present invention by more than preferred embodiment
Implement the present invention for replacement/of the invention or combination, will be apparent from for those skilled in the art, and
Among the present invention.
Claims (3)
1.A kind of polyadenylic-polyuridylic acid parenteral solution for animals, which is characterized in thatThe parenteral solution includes compound sodium chloride injection, also wraps
Include following component and its concentration：4-10g/LPolyadenylic-polyuridylic acid, 0.5-4g/L poly-D-lysines, 0.5-3g/L dodecyl sulphates
Sodium；
It to match is 1 that the polyadenylic-polyuridylic acid, which is polyinosinic acid and poly- cytimidine acid,:1 is prepared；
The preparation method of the compound sodium chloride injection is：33 mg of calcium chloride is taken, the water for injection of 100mL is added in, is placed in
It in 60-70 DEG C of water-bath, is stirred continuously to being completely dissolved, adds in 30 mg of 850 mg of sodium chloride and potassium chloride, continue stirring to solution
Become clarification to get.
2. polyadenylic-polyuridylic acid parenteral solution for animals as described in claim 1, which is characterized in that the parenteral solution includes compound chlorination
Sodium injection, also comprises the following components and its concentration：8g/L polyadenylic-polyuridylic acids, 4g/L poly-D-lysines, 3g/L dodecyl sulphur
Sour sodium.
3. the preparation method of the polyadenylic-polyuridylic acid parenteral solution for animals as described in claim 1-2 is any, which is characterized in that including with
Lower step：
S1, polyinosinic acid is taken, adds in the compound sodium chloride injection of 1/2 amount, be placed in 60-70 DEG C of water-bath, be stirred continuously to complete
Fully dissolved, it is spare；
S2, take poly- cytimidine acid, add in the compound sodium chloride injection of 1/2 amount, be placed in 60-70 DEG C of water-bath, be stirred continuously to
It is completely dissolved, it is spare；
S3, the liquid for obtaining S1 and S2 mix, and 10-20min are stirred with the speed of 30-40 r/min, in 60-70 DEG C of temperature
Lower reaction 40-60min keeps the speed stirring of 30-40 r/min in reaction process, obtains reaction solution；
S4, poly-D-lysine and lauryl sodium sulfate are added in into the reaction solution described in S3, continues stirring to being completely dissolved, treat
Solution temperature is down to room temperature, with 0.20-0.45 μm of filter filtration sterilization, be dispensed into the ampoule bottle of 5mL to get.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1031325A (en) * | 1987-08-12 | 1989-03-01 | Hem研究公司 | The Topically active compositions of double stranded RNAS |
| CN1035050A (en) * | 1987-07-17 | 1989-08-30 | Hem研究公司 | Double chain rna correction circulation immunity compound and mononuclear cell function disorder |
| CN103599071A (en) * | 2013-11-08 | 2014-02-26 | 杭州美亚药业有限公司 | Preparation method of polyinosinic-polycytidylic acid dry powder |
| CN104434784A (en) * | 2014-11-15 | 2015-03-25 | 成都天台山制药有限公司 | Medicinal composition of polyinosinic cell injection and preparation method thereof |
| CN104706580A (en) * | 2015-03-25 | 2015-06-17 | 肇庆大华农生物药品有限公司 | Veterinary interferon inducer and preparation method thereof |
-
2015
- 2015-12-22 CN CN201510965952.1A patent/CN105434341B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1035050A (en) * | 1987-07-17 | 1989-08-30 | Hem研究公司 | Double chain rna correction circulation immunity compound and mononuclear cell function disorder |
| CN1031325A (en) * | 1987-08-12 | 1989-03-01 | Hem研究公司 | The Topically active compositions of double stranded RNAS |
| CN103599071A (en) * | 2013-11-08 | 2014-02-26 | 杭州美亚药业有限公司 | Preparation method of polyinosinic-polycytidylic acid dry powder |
| CN104434784A (en) * | 2014-11-15 | 2015-03-25 | 成都天台山制药有限公司 | Medicinal composition of polyinosinic cell injection and preparation method thereof |
| CN104706580A (en) * | 2015-03-25 | 2015-06-17 | 肇庆大华农生物药品有限公司 | Veterinary interferon inducer and preparation method thereof |
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