CN108578417B - Compound sulfachlorpyridazine sodium dry suspension and preparation method thereof - Google Patents
Compound sulfachlorpyridazine sodium dry suspension and preparation method thereof Download PDFInfo
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- CN108578417B CN108578417B CN201810004452.5A CN201810004452A CN108578417B CN 108578417 B CN108578417 B CN 108578417B CN 201810004452 A CN201810004452 A CN 201810004452A CN 108578417 B CN108578417 B CN 108578417B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/63—Compounds containing para-N-benzenesulfonyl-N-groups, e.g. sulfanilamide, p-nitrobenzenesulfonyl hydrazide
- A61K31/635—Compounds containing para-N-benzenesulfonyl-N-groups, e.g. sulfanilamide, p-nitrobenzenesulfonyl hydrazide having a heterocyclic ring, e.g. sulfadiazine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/38—Cellulose; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
Abstract
The invention discloses a compound sulfachlorpyridazine sodium dry suspension and a preparation method thereof, and belongs to the technical field of veterinary drug preparations. Every 100kg of dry suspension contains: 60-70kg of sulfachlorpyridazine sodium, 10-15kg of trimethoprim, 0.02-0.05kg of suspending agent, 0.6-0.8kg of surfactant and 14.15-29.38kg of filling auxiliary material. The high-efficiency suspending agent adopted by the invention is one or two combinations of xanthan gum and hydroxypropyl methyl cellulose, the surfactant is one or two combinations of sodium lauryl sulfate and N-hexadecyl-N-ethyl morpholinyl ethyl sodium sulfate, and the filling auxiliary material is one or two combinations of maltodextrin, anhydrous glucose and water-soluble starch. The compound sulfachlorpyridazine sodium dry suspension has good suspension stability, and the prepared suspension can be uniform for a long time without layering. In clinical use, the medicament is uniformly administered no matter mixed with materials or drunk water, has good synchronous absorption effect, less medicament residue, short drug withdrawal period and convenient use, and can reduce the culture cost to the maximum extent.
Description
Technical Field
The invention relates to the field of veterinary drug preparations, and in particular relates to a compound sulfachlorpyridazine sodium dry suspension and a preparation method thereof.
Background
The sulfachlorpyridazine sodium is tasteless light yellow crystal powder in raw material property, is easy to dissolve in water, has a chemical name of N- (6-chloro-pyridazinyl) -4-aminobenzenesulfonamide sodium salt, and is one of the most important sulfanilamide medicaments. Has good curative effect on livestock and poultry infected by escherichia coli, salmonella and pasteurella, as well as coccidia such as toxoplasma of protozoa and the like. Research shows that trimethoprim has obvious synergistic effect on sulfachlorpyridazine sodium, is the most common synergist applied to sulfonamides, and has the characteristics of broad spectrum, high efficiency and low toxicity. Therefore, the sulfachlorpyridazine sodium and trimethoprim are compounded into a preparation according to a certain proportion, and the preparation is an ideal anti-infective medicament for livestock and poultry. However, trimethoprim is almost insoluble in water, and the synergistic effect of trimethoprim is seriously influenced. In the production of the preparation, the trimethoprim is dissolved by glacial acetic acid to prepare the compound sulfachlorpyridazine sodium, the problem of water solubility of trimethoprim is solved to a certain extent, however, the acetic acid residue is difficult to eliminate, the solubility of the sulfachlorpyridazine sodium in water is reduced, and the product acetic acid smell is heavy, so that the drinking water and the ingestion of animals are influenced. In addition, trimethoprim is treated by adopting a betacyclodextrin inclusion technology, the water solubility of the trimethoprim is slightly improved, but the trimethoprim is difficult to be considered as ideal and cannot meet the requirement of practical application.
The most common measure adopted for solving the problem of limited application of the insoluble medicament at present is to prepare a dry suspension, which means that the insoluble medicament and proper auxiliary materials are prepared into powder or granular materials, and the powder or granular materials can be dispersed into a suspension for oral liquid preparation by adding water and shaking when the dry suspension is used. The sedimentation speed of the drug particles of the dry suspension and the stability of the system are the most important embodiments of the suspension effect of the dry suspension. According to Stokes' law, the settling speed of the drug particles depends on the density, viscosity and particle size of the solvent, and the settling speed of the particles is proportional to the density and viscosity of the solvent and inversely proportional to the particle size of the suspension particles. Methods to increase the density and viscosity of the vehicle have limited effect on slowing the rate of microparticle sedimentation and, in many cases, adjust the particle size of the dispersed phase to maintain the physical stability of the suspension. The particle size is reduced by one time, and the sedimentation velocity is reduced by 4 times. When the particle size is reduced to the point where brownian motion is gradually apparent until the particle size is counteracted to drop and the stable state is maintained, the common dry suspension product cannot achieve the effect.
The invention emphasizes on solving the problem of drug granularity of the dry suspension, and simultaneously, the dry suspension is matched with the suspending agent and the surfactant for use, so that the dry suspension achieves the best sedimentation volume ratio, redispersibility and other rheological characteristics, and is more convenient and efficient in clinical application and wide in prospect.
Disclosure of Invention
Aiming at the problems in clinical use, the invention aims to provide a compound sulfachlorpyridazine sodium dry suspension convenient for clinical application and a technical scheme of a preparation method thereof.
The compound sulfachlorpyridazine sodium dry suspension is characterized in that 100kg of the dry suspension comprises the following components by weight:
the compound sulfachlorpyridazine sodium dry suspension is characterized in that the suspending agent is one or the combination of xanthan gum and hydroxypropyl methyl cellulose.
The compound sulfachlorpyridazine sodium dry suspension is characterized in that the surfactant is one or a combination of two of sodium lauryl sulfate and N-hexadecyl-N-ethylmorpholinyl ethyl sodium sulfate.
The compound sulfachlorpyridazine sodium dry suspension is characterized in that the filling auxiliary materials are one or two of maltodextrin, anhydrous glucose and water-soluble starch.
The compound sulfachlorpyridazine sodium dry suspension is characterized by comprising the following process steps:
(1) weighing the raw materials according to the required weight parts for later use;
(2) mixing trimethoprim and surfactant uniformly, adding water according to formula ratio, stirring uniformly, and grinding for 30min in a vertical grinding machine to obtain particle suspension with particle diameter controlled at 5-10 μm;
(3) adding suspending agent into the microparticle suspension, shearing, and passing through high pressure homogenizer to obtain microparticle emulsion;
(4) sieving the filling auxiliary materials with a 120-mesh sieve for later use;
(5) and (4) filling auxiliary materials in the step (4), uniformly mixing the auxiliary materials with the sulfachlorpyridazine sodium raw material, putting the mixture into a vertical fluidized bed, taking the particle emulsion in the step (2) as a wetting agent, and carrying out top-spraying granulation to obtain the compound sulfachlorpyridazine sodium dry suspension.
Specifically, the amount of water in the formula in the step (2) is 15-20 kg;
specifically, in the step (5), the inlet air temperature of the fluidized bed is 90-95 ℃, the outlet air temperature is 50 +/-2 ℃, and the peristaltic pump speed is 120-.
The trimethoprim raw material is strictly treated to ensure that the particle size of the trimethoprim raw material is controlled to be 5-10 mu m, and the suspension effect is excellent under the coordination of xanthan gum serving as a suspending agent, hydroxypropyl methylcellulose, sodium lauryl sulfate serving as a surfactant and the like. And the filling auxiliary material which can increase the density and viscosity of the solvent and is completely water-soluble is adopted, so that the dry suspension has good overall stability and is convenient for clinical use. Compared with the prior art, the compound sulfachlorpyridazine sodium dry suspension prepared by the invention has the following advantages:
(1) the compound sulfachlorpyridazine sodium dry suspension is produced by adopting a common fluidized bed one-step granulation technology, is simple to operate, high in safety and low in cost, and is suitable for large-scale production.
(2) The compound sulfachlorpyridazine sodium dry suspension has the advantages of good suspension effect, large volume ratio of long-time standing and sedimentation, good redispersibility, no water line blockage in the drinking water use process of poultry, and suitability for large-scale farms.
(3) The compound sulfachlorpyridazine sodium dry suspension provided by the invention is rapidly dispersed in a solvent, is synchronously absorbed in vivo, and is quick in effect.
Drawings
FIG. 1 is a graph showing the variation of sedimentation volume ratio between an experimental group and a control group;
FIG. 2 is the result of comparing the blood concentration of the experimental group with that of the control group.
Detailed Description
In order that the invention may be more readily understood and put into practical effect by the reader, reference will now be made in detail, by way of example, to the present invention. It should be understood that the following examples are only representative of the application of the present invention, and the scope of the application of the present invention is not limited thereto.
Example 1
Every 100kg of compound sulfachlorpyridazine sodium dry suspension contains: 62.5kg of sulfachlorpyridazine sodium, 12.5kg of trimethoprim, 0.03kg of xanthan gum, 0.6kg of sodium lauryl sulfate and 24.37kg of anhydrous glucose.
A preparation method of the compound sulfachlorpyridazine sodium dry suspension comprises the following steps:
(1) weighing the raw materials according to the required weight parts for later use;
(2) mixing trimethoprim and sodium lauryl sulfate uniformly, adding 16kg of water, stirring uniformly, grinding for 30min in a vertical grinding machine to obtain a particle suspension, wherein the particle size of the particles is controlled to be 5-10 mu m;
(3) adding xanthan gum into the particle suspension, uniformly shearing, and passing through a high-pressure homogenizer to obtain particle emulsion;
(4) anhydrous glucose as filling auxiliary material is sieved by a 120-mesh sieve for later use;
(5) and (3) uniformly mixing the anhydrous glucose and the sulfachlorpyridazine sodium raw material in the step (4), putting into a vertical fluidized bed, taking the particle emulsion in the step (2) as a wetting agent, and carrying out top-spraying granulation to obtain the compound sulfachlorpyridazine sodium dry suspension. The air inlet temperature of the fluidized bed is 90-95 ℃, the air outlet temperature is 50 +/-2 ℃, and the peristaltic pump speed is 120-150 ml/min.
Example 2
Every 100kg of compound sulfachlorpyridazine sodium dry suspension contains: 62.5kg of sulfachlorpyridazine sodium, 12.5kg of trimethoprim, 0.04kg of hydroxypropyl methyl cellulose, 0.8kg of N-hexadecyl-N-ethyl morpholinyl ethyl sodium sulfate and 24.16kg of maltodextrin.
A preparation method of the compound sulfachlorpyridazine sodium dry suspension comprises the following steps:
(1) weighing the raw materials according to the required weight parts for later use;
(2) mixing trimethoprim and methyl oleate uniformly, adding 16kg of water, stirring uniformly, and grinding for 30min in a vertical grinding machine to obtain a particle suspension, wherein the particle size of the particles is controlled to be 5-10 mu m;
(3) adding hydroxypropyl methyl cellulose into the particle suspension, uniformly shearing, and passing through a high-pressure homogenizer to obtain particle emulsion;
(4) sieving maltodextrin as stuffing material with 120 mesh sieve;
(5) and (3) uniformly mixing the maltodextrin and the sulfachlorpyridazine sodium raw material in the step (4), putting into a vertical fluidized bed, taking the particle emulsion in the step (2) as a wetting agent, and carrying out top-spraying granulation to obtain the compound sulfachlorpyridazine sodium dry suspension. The air inlet temperature of the fluidized bed is 90-95 ℃, the air outlet temperature is 50 +/-2 ℃, and the peristaltic pump speed is 120-150 ml/min.
Example 3
Every 100kg of compound sulfachlorpyridazine sodium dry suspension contains: 65kg of sulfachlorpyridazine sodium, 13kg of trimethoprim, 0.02kg of xanthan gum and hydroxypropyl methyl cellulose respectively, 0.3kg of sodium lauryl sulfate and 0.36 kg of N-hexadecyl-N-ethyl morpholinyl ethyl sodium sulfate respectively and 21.36kg of maltodextrin.
A preparation method of the compound sulfachlorpyridazine sodium dry suspension comprises the following steps:
(1) weighing the raw materials according to the required weight parts for later use;
(2) mixing trimethoprim with sodium lauryl sulfate and N-hexadecyl-N-ethyl morpholinyl ethyl sodium sulfate, adding 16kg of water, stirring, and grinding in a vertical grinding machine for 30min to obtain particle suspension with particle diameter of 5-10 μm;
(3) adding xanthan gum and hydroxypropyl methyl cellulose into the microparticle suspension, shearing uniformly, and passing through a high-pressure homogenizer to obtain microparticle emulsion;
(4) sieving maltodextrin as stuffing material with 120 mesh sieve;
(5) and (3) uniformly mixing the maltodextrin and the sulfachlorpyridazine sodium raw material in the step (4), putting into a vertical fluidized bed, taking the particle emulsion in the step (2) as a wetting agent, and carrying out top-spraying granulation to obtain the compound sulfachlorpyridazine sodium dry suspension. The air inlet temperature of the fluidized bed is 90-95 ℃, the air outlet temperature is 50 +/-2 ℃, and the peristaltic pump speed is 120-150 ml/min.
The compound sulfachlorpyridazine sodium dry suspension prepared by the invention is compared with a certain marketed product in the aspects of dispersion speed, sedimentation volume ratio and redispersibility in water.
Experimental groups: the compound sulfachlorpyridazine sodium dry suspension prepared in the embodiment 1 of the invention.
Control group: some market-sold compound sulfachlorpyridazine sodium product.
The test method comprises the following steps:
(1) preparing suspensions of the drugs of the experimental group and the control group according to 10 times of the dosage of 32mg (calculated by products) per kg of body weight of the clinical chicken;
(2) two 250ml beakers, labeled experimental and control, respectively, were prepared, and two 100ml test tubes, labeled experimental and control, respectively, were prepared.
(3) Weighing 320mg of the drug products of the experimental group and the control group respectively, putting the drug products into corresponding beakers, adding 100ml of tap water (25 +/-2 ℃), fully stirring the mixture, and recording the time required for the drug to be completely dispersed into a suspension state;
(4) the suspended medicines are respectively poured into corresponding test tubes, and the sedimentation volume ratio P is observed and recorded as H/H0(H: the volume after sedimentation and the volume before sedimentation of H0) every 4H;
(5) after 24h the tubes were inverted and the number of inversions required for resuspension of the experimental and control drug was recorded.
The experimental results are as follows:
TABLE 1 results of comparison of suspension effects between the experimental group and the control group
And (4) experimental conclusion:
(1) the same amount of the drug is dispersed in water by stirring, the experimental group can completely disperse in only 20s, while the control group needs 2 min. The compound sulfachlorpyridazine sodium dry suspension is obviously superior in dispersibility, better meets the characteristics of the dry suspension and is more convenient to use clinically.
(2) The sedimentation volume ratio of the experimental group is slowly reduced and more stable in the standing time of the suspension from 4h to 24h, the sedimentation volume ratio still reaches 0.902 after 24h, while the sedimentation volume ratio of the control group is rapid, and the sedimentation volume ratio is 0.412 and is unstable after 24 h.
(3) After the suspension is kept stand for 24 hours, the shaking times of the experimental group and the control group are respectively 8 times and 23 times due to redispersion, which shows that the compound sulfachlorpyridazine sodium dry suspension has good redispersibility and is more favorable for clinical use.
In order to better illustrate the absorption utilization rate and the drug effect of the compound sulfachlorpyridazine sodium dry suspension and a commercially available product, clinical tests are specially carried out in certain pig farms in Zhejiang.
The test method comprises the following steps:
(1) selecting 30 piglets which are 15 days old and have white scour symptom, and averagely dividing into A, B groups, wherein each group has 15 piglets;
(2) the group A is administrated by using the medicine of the invention in the drinking water, is a test group, the group B is administrated by using a certain marketed medicine in the drinking water, is a control group, and the addition amount is 200g/t water (calculated by products). After administration, 3 piglets are selected from each group, blood is collected and blood concentration is measured at 0.5h, 1h, 2h, 4h, 6h, 8h, 10h and 12h, and the average value of 3 piglets in each group is calculated.
(3) A, B two groups of piglets are administered with water continuously for 5 days, and the cure rate is counted 3 days and 6 days after administration.
And (3) test results:
TABLE 2 comparison of cure rates of porcine white diarrhea
And (4) test conclusion:
as can be seen from the results of comparing the blood concentration of the experimental group with that of the control group shown in figure 2, the peak area of the drug in the embodiment 1 of the invention is obviously larger than that of the marketed product, and the sulfachlorpyridazine sodium and the trimethoprim simultaneously reach the peak value of the blood, so that the drug has good effect taking synchronism. Meanwhile, the medicine of the example 1 has short peak reaching time and quick response.
Table 2 shows that the medicine in the example 1 has short cure period and high cure rate for the white diarrhea of pigs, and is obviously superior to the marketed products.
Claims (2)
1. The compound sulfachlorpyridazine sodium dry suspension is characterized by comprising the following components in parts by weight of 100 kg:
the suspending agent is one or the combination of xanthan gum and hydroxypropyl methylcellulose;
the xanthan gum and the hydroxypropyl methyl cellulose are food grade, and the absolute viscosity is more than 10 ten thousand mPa & s;
the surfactant is one or two of sodium lauryl sulfate and N-hexadecyl-N-ethyl morpholinyl ethyl sodium sulfate;
the filling auxiliary material is one or two of maltodextrin, anhydrous glucose and water-soluble starch;
the dry suspension is a microparticle suspension, and the particle size of microparticles is controlled to be 5-10 mu m.
2. The preparation method of the compound sulfachlorpyridazine sodium dry suspension of any claim 1, is characterized by comprising the following steps:
(1) weighing the raw materials according to the required weight parts for later use;
(2) mixing trimethoprim and surfactant uniformly, adding water according to the formula amount, stirring uniformly, and grinding for 30min in a vertical sand mill to obtain particle suspension with the particle diameter controlled at 5-10 μm;
(3) adding suspending agent into the microparticle suspension, shearing, and passing through high pressure homogenizer to obtain microparticle emulsion;
(4) sieving the filling auxiliary materials with a 120-mesh sieve for later use;
(5) filling auxiliary materials in the step (4), uniformly mixing the auxiliary materials with the sulfachlorpyridazine sodium raw material, putting the mixture into a vertical fluidized bed, taking the particle emulsion in the step (3) as a wetting agent, and carrying out top-spraying granulation to obtain the compound sulfachlorpyridazine sodium dry suspension;
the amount of water in the formula in the step (2) is 15-20 kg;
in the step (5), the air inlet temperature of the fluidized bed is 90-95 ℃, the air outlet temperature is 50 +/-2 ℃, and the peristaltic pump speed is 120-150 ml/min.
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| CN101416973A (en) * | 2007-10-22 | 2009-04-29 | 洛阳普莱柯生物工程有限公司 | Preparation method of novel composite anti-coccidium, antibiotic preparation |
| CN101468025A (en) * | 2007-12-26 | 2009-07-01 | 天津瑞普生物技术集团有限公司 | Soluble powder for preventing and treating colibacillosis of fowl |
| WO2010143207A1 (en) * | 2009-06-11 | 2010-12-16 | Rubicon Research Private Limited | Taste-masked oral formulations of influenza antivirals |
| CN102247386A (en) * | 2011-05-26 | 2011-11-23 | 河南牧翔动物药业有限公司 | Soluble powder used for treating poultry coccidiosis |
| CN102824350A (en) * | 2012-08-20 | 2012-12-19 | 安徽新和成皖南药业有限公司 | Compound sulfamethoxazole dry suspension and preparation method thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416973A (en) * | 2007-10-22 | 2009-04-29 | 洛阳普莱柯生物工程有限公司 | Preparation method of novel composite anti-coccidium, antibiotic preparation |
| CN101468025A (en) * | 2007-12-26 | 2009-07-01 | 天津瑞普生物技术集团有限公司 | Soluble powder for preventing and treating colibacillosis of fowl |
| WO2010143207A1 (en) * | 2009-06-11 | 2010-12-16 | Rubicon Research Private Limited | Taste-masked oral formulations of influenza antivirals |
| CN102247386A (en) * | 2011-05-26 | 2011-11-23 | 河南牧翔动物药业有限公司 | Soluble powder used for treating poultry coccidiosis |
| CN102824350A (en) * | 2012-08-20 | 2012-12-19 | 安徽新和成皖南药业有限公司 | Compound sulfamethoxazole dry suspension and preparation method thereof |
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Denomination of invention: A compound sulfachlorpyridazine sodium dry suspension and its preparation method Effective date of registration: 20220929 Granted publication date: 20200515 Pledgee: Zhejiang Anji Rural Commercial Bank of the West Branch of Limited by Share Ltd. Pledgor: ZHEJIANG VEGA BIO-TECHNOLOGY CO.,LTD. Registration number: Y2022330002388 |
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