CN118320063A

CN118320063A - Antishock resuscitation fluid

Info

Publication number: CN118320063A
Application number: CN202310031182.8A
Authority: CN
Inventors: 李金宝; 帖红涛; 周义萱; 曹静怡
Original assignee: Beijing Caimicrobiota Technology Co ltd
Current assignee: Beijing Caimicrobiota Technology Co ltd
Priority date: 2023-01-10
Filing date: 2023-01-10
Publication date: 2024-07-12

Abstract

The invention discloses an antishock resuscitation fluid. The antishock resuscitation fluid comprises the following components: albumin, sodium octoate and crystal liquid, wherein each 500ml of anti-shock recovery liquid contains 2.5-50 g of albumin, 1.67 g of sodium octoate, 3.0-10 g of salt in the crystal liquid and 500ml of water for injection. The invention reduces the clinical shock patient's resuscitation treatment steps from three steps to one step. Namely: the three steps of traditional ringer's solution, albumin and normal saline are directly simplified into one step: the compound albumin resuscitation fluid saves the golden time of treatment; the antishock resuscitation fluid is input into a patient at one time, so that the survival rate of the patient is improved; reducing pollution risk. Clinically, the compound albumin resuscitation fluid is very convenient to use, is easier to popularize clinically, and is beneficial to shock patients.

Description

Antishock resuscitation fluid

Technical Field

The invention belongs to the field of medicines, and particularly relates to an anti-shock resuscitation fluid.

Background

Shock (shock) is a syndrome characterized by deregulation of body nerves, humoral factors and acute microcirculation disturbance caused by various serious pathogenic factors, and directly or indirectly causes extensive damage to important visceral cells. Shock is a systemic pathological process in which under the action of strong pathogenic factors such as serious infection, trauma, blood loss, and the like, tissue blood perfusion is seriously insufficient, and effective circulating blood volume is drastically reduced, so that vital organs and cell function are metabolized and structural damage is caused to the organism (reference document: huang Pei, et al: clinical effects and significance of pre-hospital emergency treatment of traumatic shock patients [ J ]. First-aid foods and medicines, 2019, 26 (10): 96).

Shock can be divided into several categories depending on the cause: severe war wound, disaster wound, burn and scald, sepsis, upper respiratory tract obstruction, asphyxia, tension pneumothorax, and hemorrhagic shock. Among them, hemorrhagic shock (hemorrhagic shock) is the most common. Hemorrhagic shock is a shock with hypovolemia that occurs as a result of a sudden decrease in circulating blood volume caused by massive blood loss. Traumatic hemorrhagic shock is a common clinical critical syndrome, both at ordinary times and at war time. It is reported to be the first cause of death in the age range of 1 to 44 years. Previous data indicate that about 50% of war wound wounded persons are fatal due to massive blood loss (reference document ：Stern SA.et al.Effect of initially limited re-suscitation in acombined model of fluid percussion brain injury and severe uncontrolled hemorrhagic shock[J].J Neurosurg,2000,93:305314.).

Hemorrhagic shock is a common clinical critical syndrome. Liquid resuscitation is the primary treatment for hemorrhagic shock, and in existing liquid resuscitation methods, lyophilized plasma (freeze-DRIED PLASMA) and human albumin (human albumin) play a significant role (ref: zhao Xiong et al: use of lyophilized plasma and human albumin in the treatment of war wound shock. Military medicine, 2015, 39 (3): 216-219). At present, physiological saline or ringer's solution is mainly infused into the pre-hospital treatment of hemorrhagic shock, but for severe hemorrhagic shock wounded persons caused by repeated and continuous blood loss for a long time in the transportation process, the infusion of crystal solution can not effectively supplement erythrocytes, relieve tissue hypoxia, and the wounded persons are easy to develop into refractory shock, which is a main cause of death of the hemorrhagic shock wounded persons. Currently, there are three main types of antishock resuscitation fluids: crystalline fluid, colloidal fluid and lyophilized plasma.

1. Crystalline liquid:

the crystal liquid refers to sodium chloride liquid and ringer's liquid. Ringer's solution is also called compound sodium chloride, and is invented by British physiology' ringer's solution, so that it is called ringer's solution, and is actually commonly called compound sodium chloride injection. The ringer's solution contains potassium ions, calcium ions and magnesium ions in addition to sodium chloride. Ringer's solution is an isotonic electrolyte solution containing lactate and no bicarbonate, and its electrolyte concentration, pH, osmotic pressure, etc. are very similar to those of extracellular fluid, so it is also called equilibrium salt solution. Has important physiological effects on preventing and treating hypovolemic shock, has been widely used to replace lost extracellular fluid or as a vehicle for plasma substitutes to increase its therapeutic value, and can prevent chronic intravascular coagulation (DIC) to prevent shock. Belongs to a medicine for regulating water, electrolyte and acid-base balance.

Sodium lactate is added on the basis of ringer's solution to form sodium lactate ringer's solution, which is also called Hartmann's solution. And adding other components such as sodium acetate, glucose and the like on the basis of the ringer's solution to obtain the sodium acetate ringer's solution. The sodium lactate ringer's solution or the sodium acetate ringer's solution has better therapeutic shock effect than the physiological saline of sodium chloride in clinic, so the sodium lactate ringer's solution or the sodium acetate ringer's solution is widely used in clinic.

2. Gel liquid:

The colloid liquid comprises gelatin, glucose polymer, hydroxyethyl starch (high molecular blood albumin is the third generation hydroxyethyl starch injection), and human serum albumin. The colloidal fluid has strong dilatation effect, long residence time in blood vessels, and can be rapidly infused in the colloidal fluid in a short time, thereby more effectively increasing blood volume and cardiac output than the rapid infusion of the colloidal fluid. From the aspect of capacity replacement requirements, the blood volume supplement and the interstitial and intracellular fluid and electrolyte supplement are required. Numerous studies have shown that the degree of activation of neutrophils during traumatic hemorrhagic shock is related to the type of resuscitation fluid, not just to the resuscitation itself, and that activation and release of a large number of inflammatory cytokines by inflammatory cells, neutrophils, endothelial cells, etc., as well as oxygen radical damage, etc., are important causes of shock.

Compared with the conventional large-scale application of the liquid crystal resuscitation, the liquid crystal resuscitation can reduce the level of plasma volume during traumatic hemorrhagic shock, the liquid volume required by resuscitation is obviously reduced, and the lung tissue injury is reduced. In addition, the application of the hypertonic solution such as colloid liquid has obvious capacity expansion effect and the effect of avoiding ischemia reperfusion injury, overload of liquid and the like caused by the recovery of a large amount of crystal liquid.

3. Freeze-dried plasma:

The freeze-dried blood plasma is solid powder product prepared by mixing whole blood donated by healthy people according to certain proportion, inactivating virus and freeze-drying. The main components of the freeze-dried blood plasma are basically the same as fresh frozen blood plasma, and most of blood coagulation factors including albumin, immunoglobulin, fibrinogen and the like can effectively maintain the osmotic pressure of colloid in vivo, increase the circulating blood volume and supplement the blood plasma protein. The freeze-dried blood plasma is generally matched with erythrocytes for use in the early stage of severe war injury, and the two are applied according to the ratio of 1:1. By correcting clotting factor disturbances, the survival rate of wounded persons is greatly increased (ref: martinaud C, et al, the frequency frame DRIED PLASMA [ J ]. J Trauma,2011, 71 (4): 1091-1092).

The existing antishock resuscitation fluid has the problems that:

1. Crystalline liquid:

the crystal liquid mainly comprises sodium chloride physiological saline and ringer's solution. Sodium chloride physiological saline has inferior therapeutic effect on shock. Ringer's solution is deficient in that it is higher than plasma Cl ^- and less Na ⁺, and has a lower osmotic level than plasma, and patients with head trauma tend to have cerebral edema associated therewith, so it has been suggested that ringer's solution should be limited to patients at risk for cerebral edema.

For sodium lactate ringer's solution or sodium acetate ringer's solution, metabolic alkalosis can result due to rapid metabolism of lactate or acetate into CO ₂ and water, and if ringer's solution is administered in large amounts, such an alkali substitution can result in compensatory respiratory acidosis and postoperative respiratory depression.

The ringer's solution is used simply, which is easy to cause hypoalbuminemia and causes obvious adverse reaction.

The use of ringer's solution is a basic therapy to correct water and electrolyte deficiency after blood and body fluid loss because it is inexpensive, effective, well protected from kidney function, and conventional massive ringer's solution resuscitation during shock can lead to significant reperfusion injury and fluid overload, resulting in tissue edema, and can induce pulmonary edema due to dilution of plasma proteins.

2. Gel liquid:

In recent years, a relatively large number of colloid solutions are clinically used, namely high molecular blood-replacement albumin, which is a third-generation product of hydroxyethyl starch. Hydroxyethyl starch belongs to a mixture composed of a plurality of different molecular weights, has high molecular solutes, is not easy to exude from capillaries, has longer residence time in human blood vessels, can obviously promote colloid osmotic pressure, helps tissue fluid to return to the blood vessels, and plays a good dilatation effect. Clinical and animal experiments show that the polymer blood-replacing albumin injection can continuously raise the mean arterial pressure, the systolic pressure and the diastolic pressure of hemorrhagic shock; can quickly recover hematocrit of hemorrhagic shock and improve ischemia state (reference document: he Jingwei Na et al: high molecular substituted albumin for the observation of therapeutic effect of canine hemorrhagic shock: heilongjiang animal doctor, 2020 (9): 107-109: wang Xiaojing: clinical comparison of hydroxyethyl starch solution and albumin in the treatment of hemorrhagic shock patient volume: new medicine, 2007, 37 (1): 29-31). However, the macromolecular blood-replacing albumin not only has small protein content, but also can cause adverse reaction of blood transfusion; hydroxyethyl starch has a dose-dependent effect on renal function, clotting function and possible allergic reactions and is prone to hypoalbuminemia in severely burn patients after burn shock phase application.

3. Freeze-dried plasma:

In recent years, the risk of death of patients is significantly increased by the potential hazards of lyophilized plasma, including the transmission of infectious diseases such as hepatitis, aids, and the like, as well as the resulting serious complications such as acute hemolytic transfusion reactions, bacterial contamination, allergic reactions, transfusion-related acute lung injuries, and the like, in addition to blood-borne stress factors. In the case of increased capillary permeability (e.g., in sepsis), it also penetrates the interstitial space and exacerbates tissue edema (ref: jiang Shouyin et al: research on the use of a novel strategy for resuscitation in traumatic hemorrhagic shock and its mechanism [ D ]. Hangzhou: zhejiang university 2015: 67-84).

The prior antishock resuscitation fluid used clinically has complex procedures, and wastes precious gold treatment time. The first principle in pre-hospital emergency is to revive liquid in time and early.

The literature (Lin Jinbo et al: albumin combined with fluid resuscitation for the observation of changes in clearance of blood lactate and lactate in pediatric septic shock. Contemporary medicine, 2021, 27 (14): 169-170) reports that patients were first intravenously instilled with ringer's solution for 30 minutes, then with human serum albumin for 20 minutes, and then with physiological saline for 30 minutes. It is known that, because ringer's solution, human serum albumin and physiological saline are clinically used, they cannot be combined together to complete one-time instillation, which is very inconvenient clinically, and at the same time, the risk of pollution and the labor cost are brought by frequent liquid exchange. The infusion method delays precious treatment time.

The literature (Zhou Xuejian et al, influence of resuscitation fluid on traumatic shock plasma TNF- α, IL 6. Armed forces, 2011, 22 (4): 26-27) reports that traumatic hemorrhagic shock patients were treated with different antishock resuscitation fluids. Ringer's solution group, 25% human serum albumin group and hydroxyethyl starch injection group. By detecting TNF-alpha and IL6 in the lungs of patients, the following results are found: the therapeutic effect of human serum albumin and hydroxyethyl starch is better than that of ringer's solution. Meanwhile, the ringer's solution also has certain curative effect in traumatic hemorrhagic shock. The reported disadvantage is that three groups were not cross tested.

The clinical efficacy of plasma exchange using albumin and/or ringer's solution (Li Cuifang et al, J.International urinary System, 2018, 38 (3): 467-470) reported that authors used ringer's solution and/or human serum albumin for plasma exchange assays. As a result, it was found that plasma exchange treatment was performed by substituting part of plasma with ringer's solution+5% human serum albumin, and a preferable result was obtained. This document is mainly used in connection with plasmapheresis therapy and has not been used for anti-shock resuscitation fluids.

Human albumin (Alb) is the most abundant protein in plasma, accounting for 55% -60% of total protein in plasma, and the osmotic pressure produced by the human albumin (Alb) accounts for 80% of the osmotic pressure of plasma colloid. Albumin has a relative molecular mass of (65-68) ×10 ³, an isoelectric point of 4.7-4.9, a half-life period of about 15d, and has physiological functions of maintaining plasma colloid osmotic pressure, combining and transporting small molecular substances in blood, resisting oxidation, anticoagulation, and immunoregulation in muscle, liver and kidney catabolism (refer to ：Friedman AN et al.Reassessment of albumin as an nutritional marker in kidney disease[J].J Am Soc Nephrol,2010,21(2):223-230). albumin) ：Ha CE,et al.Novel insights into the pleiotropic effects of human serum albumin in health and disease[J].Biochim Biophys Acta,2013,1830(12):5486-5493).

Maintaining plasma colloid osmotic pressure is one of the most important physiological functions of albumin. The colloid osmotic pressure is in direct proportion to the number of macromolecules in the solution, and the albumin has higher relative molecular mass and slower speed of penetrating through the membrane compared with salts and moisture, so that the colloid osmotic pressure of the albumin is balanced with the static pressure of capillary vessels, thereby maintaining normal blood volume. In addition, albumin has a certain hydration effect. 1g albumin may retain 18ml water, while 1g plasma protein may retain 15ml water. From this, it was estimated that the ability of 10g albumin to retain moisture in the circulation corresponds to 200ml plasma. Therefore, albumin is effectively supplemented in early stage, the osmotic pressure of the intravascular colloid is increased, the interstitial resorption is facilitated, and the effective blood volume is increased. Human serum albumin can regulate the dynamic balance of water between tissues and blood vessels, and is one of important medicines for preventing and treating shock (refer to literature ：Bansal ML,et al.Relative survival benefit and morbidity with fluids in severe sepsis a network meta-analysis of alternative therapies[J].Curr Drug Saf,2013,8(4):236-245).. Infusion of human serum albumin can quickly suck water into blood circulation to maintain blood volume and heart beat volume. Severe bleeding on combat wounds, especially when the blood volume loss is more than 50%, besides supplementing whole blood, the human serum albumin needs to be infused to maintain the plasma protein more than 52g/L so as to ensure the plasma colloid osmotic pressure.

Albumin plays a vital role and role as an anti-shock resuscitation fluid. Albumin alone, however, has the following drawbacks as an anti-resuscitation fluid:

1) Albumin in anti-shock resuscitation fluids, the pH of the solution is maintained less stable than ringer's solution. Ringer's solution has an advantage in terms of the pH buffering capacity of the solution because ringer's solution, either lactate or acetate, contains a pH buffer, whereas albumin solutions lack such a buffer system.

2) Albumin has a certain osmotic pressure in anti-shock resuscitation fluid, but the osmotic pressure of albumin itself is not high because albumin acts as a colloid. If albumin is diluted from 25% to 5%, the osmotic pressure drops sharply, and is insufficient in maintaining the osmotic pressure of the shock patient, requiring ringer's solution or physiological saline to regulate the osmotic pressure.

Disclosure of Invention

The invention aims to provide an anti-shock resuscitation fluid.

The anti-shock resuscitation fluid provided by the invention, namely, the compound albumin resuscitation fluid, comprises the following components:

albumin, sodium octoate and crystal liquid,

Each 500ml of the anti-shock resuscitation fluid contains 2.5-50 g (preferably 5-40 g, more preferably 25 g) of albumin, 1.67 g of sodium octoate, 3.00 g-10.00 g (preferably 3.13-8.951 g) of salt in the crystal fluid and 500ml of water for injection.

Wherein the albumin is at least one of blood plasma-derived human serum albumin and gene recombination-derived human serum albumin;

The crystal liquid can be any one of sodium chloride physiological saline and ringer's solution.

Specifically, the crystalline fluid may be selected from: sodium lactate ringer's solution and sodium acetate ringer's solution.

Preferably, the anti-shock resuscitation fluid consists of the formulation of any one of the following 1) -4):

1) Each 500ml contains:

2.5-50 g of plasma source human serum albumin;

sodium octoate 1.67g

Sodium lactate 1.55g

Sodium chloride 1.33g

Potassium chloride 0.15g

Calcium chloride dihydrate 0.10g

500Ml of water for injection;

2) Each 500ml contains:

2.5-50 g of plasma source human serum albumin;

sodium octoate 1.67g

Sodium chloride 1.516g

Potassium chloride 0.150g

Magnesium chloride 0.102g

Sodium citrate 0.294g

1.702G of sodium acetate

Calcium gluconate 0.336g

Anhydrous dextrose 5.001g

500Ml of water for injection;

3) Each 500ml contains:

2.5-50 g of gene recombination source human serum albumin;

sodium octoate 1.67g

Sodium lactate 1.55g

Sodium chloride 1.33g

Potassium chloride 0.15g

Calcium chloride dihydrate 0.10g

500Ml of water for injection;

4) Each 500ml contains:

2.5-50 g of gene recombination source human serum albumin;

sodium octoate 1.67g

Sodium chloride 1.516g

Potassium chloride 0.150g

Magnesium chloride 0.102g

Sodium citrate 0.294g

1.702G of sodium acetate

Calcium gluconate 0.336g

Anhydrous dextrose 5.001g

500Ml of water for injection.

The antishock resuscitation fluid is prepared by a method comprising the following steps:

dissolving other raw materials except albumin in water for injection, filtering to remove impurities, and sterilizing to obtain solution; adding albumin injection, ultrafiltering, concentrating, and preserving at low temperature.

Compared with the prior art, the invention has the following advantages:

1) Shortening the treatment time: traumatic hemorrhagic shock, during the rescue process, urgency and rapidity are key points for rescuing patients from resuscitation in golden time. The invention reduces the steps of resuscitation treatment of shock patients from three steps to one step. Namely: the three steps of traditional ringer's solution, albumin and normal saline are directly simplified into one step: the compound albumin resuscitation fluid saves the golden time for treatment.

2) The survival rate is improved: traumatic hemorrhagic shock, the first few hours are critical time points. The compound albumin resuscitation fluid is infused into a patient at one time, so that the survival rate of the patient is improved.

3) Reducing pollution risk: the repeated injection of different kinds of medicine to patient may result in medicine pollution and patient infection. The invention reduces three steps into one step, and the pollution risk is greatly reduced.

4) Clinically, the compound albumin resuscitation fluid is very convenient to use, is easier to popularize clinically, and is beneficial to shock patients.

The invention mixes albumin and crystal liquid for composite use. The crystal liquid is not only a buffer solution of albumin, but also plays a role in stabilizing albumin; the crystalloid fluid is also an antishock resuscitation fluid. Simultaneously, the crystal liquid and albumin are simultaneously input into the wounded body, so that the survival rate of the shock patient is improved, and the treatment time of the shock patient is shortened.

Detailed Description

The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.

The sources of raw materials in the following examples:

1) Plasma-derived human serum albumin injection: the manufacturer is Shanghai Laiyi. Specification of: 12.5g,50 ml/bottle. Protein concentration 25%.

2) Recombinant human serum albumin injection: the manufacturer is Wuhan grass. Specification of: 12.5g,50 ml/bottle. Protein concentration 25%.

3) Sodium L-lactate solution: the manufacturer is a national pharmaceutical group chemical reagent company. Specification of: 60% solution (Walki). And (3) packaging: 25 g.

4) Sodium chloride: the manufacturer is the company of Simanyue medical science and technology. Specification of: raw material medicines. And (3) packaging: 500 g.

5) Potassium chloride: the manufacturer is a national pharmaceutical group chemical reagent company. Specification of: GR. And (3) packaging: 500 g.

6) Calcium chloride dihydrate: the manufacturer was Sigma. Specification of: sigma 99+%. And (3) packaging: 500 g.

7) Anhydrous magnesium chloride: the manufacturer is a national pharmaceutical group chemical reagent company. Specification of: 99.9% powder (Wobbe). And (3) packaging: 500 g.

8) Trisodium citrate dihydrate: the manufacturer is a national pharmaceutical group chemical reagent company. Specification of: GR (Shanghai test) is more than or equal to 99.5 percent. And (3) packaging: 500 g.

9) Sodium acetate trihydrate: the manufacturer is a national pharmaceutical group chemical reagent company. Specification of: AR (Shanghai test) is more than or equal to 99.0%. And (3) packaging: 500 g.

10 Calcium gluconate: the manufacturer is a national pharmaceutical group chemical reagent company. Specification of: 99% (Wobbe). And (3) packaging: 500 g.

11 Anhydrous dextrose: the manufacturer is Shanxi jin ocean pharmaceutical excipients Limited company. Specification of: pharmaceutical grade. And (3) packaging: 500 g.

12 Sodium octoate): the manufacturer was Sigma. Specification of: 99% (CAPILLARY GC) or more. And (3) packaging: 500 g.

13 Water for injection): the manufacturer is Shandong Qi Du pharmaceutical Co. Specification of: sterilizing the water for injection. And (3) packaging: 500ml.

The instrument and equipment used are as follows:

1) 30kDa ultrafiltration membrane package: the manufacturer is Millipore company. Specification of: biomax 30kda.

2) An electronic balance: the manufacturer is Metrehler company. Model: XPR106DUH/AC.

3) UV/visible spectrophotometer: the manufacturer is Metrehler company.

Example 1 preparation of Compound Albumin resuscitation fluid A

In the raw materials used in the test, human serum albumin from plasma was purchased from Shanghai Laiyer and packaged in glass bottles of 12.5 g/50 ml and 25% albumin concentration. The sodium L-lactate solution was purchased from national pharmaceutical systems chemical reagent Co., ltd, and was packaged as a 60% strength solution (Wobbe). Sodium chloride is purchased from the company of Simarouba medical science and technology, and raw materials with package specifications are packaged. Potassium chloride was purchased from Guogou chemical Co., ltd, and was packaged to a GR specification. Calcium chloride dihydrate was purchased from Sigma, package size Sigma 99+%. The water for injection is purchased from Shandong Mimo pharmaceutical Co., ltd, and the water for injection is sterilized in package specification.

The specific formula is as follows:

Formula I: compound albumin resuscitating liquid A

Each 500ml contains:

plasma-derived human serum albumin 25 g

Sodium octoate 1.67g

Sodium lactate 1.55g

Sodium chloride 1.33g

Potassium chloride 0.15g

Calcium chloride dihydrate 0.10g

500Ml of water for injection

According to the above formula, each raw material (except for blood plasma source human serum albumin) is accurately weighed by an electronic balance, placed in a clean beaker after sterilization, after weighing, sterilized injection water is added, fully stirred until the raw materials are completely dissolved, and then impurities are removed by a1 μm filter and a 0.45 μm filter respectively, and filtrate is collected. Then, the mixture was filtered through a 0.22 μm sterilization filter, and the filtrate was collected for use.

Since the plasma-derived human serum albumin injection contains small molecules such as salt ions, it is necessary to perform the test by replacing the plasma-derived human serum albumin injection with a salt solution of formula I. The present invention employs a 30Kda ultrafiltration membrane pack (model number Biomax30Kda, manufactured by Millipore Co.). A30 Kda ultrafiltration membrane pack, according to manufacturer's instructions, entraps compounds having a molecular weight greater than 30Kda, while compounds having a molecular weight less than 30Kda are filtered out. Therefore, the method can not only keep the human serum albumin (the molecular weight is 65-68 Kda), but also filter out small molecules such as salt ions in the human serum albumin injection. In addition, the 30Kda ultrafiltration membrane bag also has a concentration function, and can be concentrated to 1/5-1/20 of the original volume at most, so that the concentration of protein is improved.

The test uses the ultrafiltration membrane package produced by Millopre company to carry out ultrafiltration and concentration.

According to the product operating manual provided by Millipore company of the manufacturer. The Biomax 30Kda ultrafiltration membrane bag was properly mounted. The ultrafiltration membrane pack was placed on ice for pre-cooling for 30 minutes. The liquid peristaltic pump was turned on and the membrane pack was rinsed with sterile water for injection for 3 pack volumes. Then, 5 membrane package volumes were equilibrated with albumin-free compound albumin resuscitation fluid a. And adding the blood plasma source human albumin into the albumin-free compound albumin resuscitation fluid A, and uniformly mixing. After 5 membrane-pack volumes were replaced, concentration was performed, and the concentration of human serum albumin was simultaneously detected with a UV/visible spectrophotometer. Concentration was stopped until the human serum albumin concentration reached 25 g/500 ml. Collecting concentrated solution, and placing in a refrigerator at 2-8deg.C for use.

Example 2 preparation of Compound Albumin resuscitation fluid B

In the raw materials used in the test, human serum albumin from plasma was purchased from Shanghai Laiyer and packaged in glass bottles of 12.5 g/50 ml and 25% albumin concentration. Sodium chloride is purchased from the company of Simarouba medical science and technology, and raw materials with package specifications are packaged. Potassium chloride was purchased from Guogou chemical Co., ltd, and was packaged to a GR specification. Anhydrous magnesium chloride was purchased from national pharmaceutical chemicals limited, package size 99.9% powder (wokak). Trisodium citrate dihydrate is purchased from national pharmaceutical group chemical reagent company, and has a package specification GR (Shanghai test) of 99.5% or more. Sodium acetate trihydrate is purchased from national pharmaceutical group chemical reagent company, and the packaging specification AR (Shanghai test) is more than or equal to 99.0%. Calcium gluconate is purchased from national pharmaceutical group chemical reagent company, inc., package size 99% (Woki). Anhydrous glucose is purchased from Shanxi jin ocean pharmaceutical excipients, inc., and is packaged into pharmaceutical grade. The water for injection is purchased from Shandong Mimo pharmaceutical Co., ltd, and the water for injection is sterilized in package specification.

The specific formula is as follows:

and the formula II: compound albumin resuscitating liquid B

Each 500ml contains:

plasma-derived human serum albumin 25 g

Sodium octoate 1.67g

Sodium chloride 1.516g

Potassium chloride 0.150g

Magnesium chloride 0.102g

Sodium citrate 0.294g

1.702G of sodium acetate

Calcium gluconate 0.336g

Anhydrous dextrose 5.001g

500Ml of water for injection

Since the plasma-derived human serum albumin injection contains small molecules such as salt ions, it is necessary to perform the test by replacing the plasma-derived human serum albumin injection with a salt solution of the second formula. The present invention employs a 30Kda ultrafiltration membrane pack (model number Biomax30Kda, manufactured by Millipore Co.). A30 Kda ultrafiltration membrane pack, according to manufacturer's instructions, entraps compounds having a molecular weight greater than 30Kda, while compounds having a molecular weight less than 30Kda are filtered out. Therefore, the method can not only keep the human serum albumin (the molecular weight is 65-68 Kda), but also filter out small molecules such as salt ions in the human serum albumin injection. In addition, the 30Kda ultrafiltration membrane bag also has a concentration function, and can be concentrated to 1/5-1/20 of the original volume at most, so that the concentration of protein is improved.

The operation was performed according to the product operation manual provided by Millipore company of the manufacturer. The Biomax30Kda ultrafiltration membrane bag was properly mounted. The ultrafiltration membrane pack was placed on ice for pre-cooling for 30 minutes. The liquid peristaltic pump was turned on and the membrane pack was rinsed with sterile water for injection for 3 pack volumes. Then, 5 membrane package volumes were equilibrated with albumin-free compound albumin resuscitation fluid B. Adding the plasma-derived human serum albumin injection into the albumin-free compound albumin resuscitator B, uniformly mixing, replacing 5 membrane package volumes, concentrating, and simultaneously detecting the concentration of human serum albumin by using a UV/visible spectrophotometer. Concentration was stopped until the human serum albumin concentration reached 25 g/500 ml. Collecting concentrated solution, and placing in a refrigerator at 2-8deg.C for use.

Example 3 preparation of Compound Albumin resuscitation fluid C

In the raw materials adopted in the test, the human serum albumin from the gene recombination source is purchased from Wuhan Gramineae, the packaging specification is 12.5 g/50 ml glass bottle, and the albumin concentration is 25%. The sodium L-lactate solution was purchased from national pharmaceutical systems chemical reagent Co., ltd, and was packaged as a 60% strength solution (Wobbe). Sodium chloride is purchased from the company of Simarouba medical science and technology, and raw materials with package specifications are packaged. Potassium chloride was purchased from Guogou chemical Co., ltd, and was packaged to a GR specification. Calcium chloride dihydrate was purchased from Sigma, package size Sigma 99+%. The water for injection is purchased from Shandong Mimo pharmaceutical Co., ltd, and the water for injection is sterilized in package specification.

The specific formula is as follows:

and the formula III: compound albumin resuscitating liquid C

Each 500ml contains:

2.5-50 g of gene recombination source human serum albumin;

Sodium octoate 1.67g

Sodium lactate 1.55g

Sodium chloride 1.33g

Potassium chloride 0.15g

Calcium chloride dihydrate 0.10g

500Ml of water for injection

According to the above formula, each raw material (excluding the human serum albumin from the gene recombination source) is accurately weighed by an electronic balance, placed in a clean beaker after sterilization, after weighing, sterilized water for injection is added, fully stirred until completely dissolved, and then impurities are removed by a1 μm filter and a 0.45 μm filter respectively, and filtrate is collected. Then, the mixture was filtered through a 0.22 μm sterilization filter, and the filtrate was collected for use.

Because the human serum albumin injection from the gene recombination source contains small molecules such as salt ions, the test needs to be carried out by replacing the small molecules with a salt solution in a formula III. The present invention employs a 30Kda ultrafiltration membrane pack (model number Biomax 30Kda, manufactured by Millipore Co.). A30 Kda ultrafiltration membrane pack, according to manufacturer's instructions, entraps compounds having a molecular weight greater than 30Kda, while compounds having a molecular weight less than 30Kda are filtered out. Therefore, the method can not only keep the human serum albumin (the molecular weight is 65-68 Kda), but also filter out small salt ion molecules in the human serum albumin injection. In addition, the 30Kda ultrafiltration membrane bag also has a concentration function, and can be concentrated to 1/5-1/20 of the original volume at most, so that the concentration of protein is improved.

The operation was performed according to the product operation manual provided by Millipore company of the manufacturer. The Biomax30Kda ultrafiltration membrane bag was properly mounted. The ultrafiltration membrane pack was placed on ice for pre-cooling for 30 minutes. The liquid peristaltic pump was turned on and the membrane pack was rinsed with sterile water for injection for 3 pack volumes. Then, 5 membrane package volumes were equilibrated with albumin-free compound albumin resuscitation fluid C. Adding the gene recombinant source human serum albumin injection into the albumin-free compound albumin resuscitator C, uniformly mixing, replacing 5 membrane package volumes, concentrating, and simultaneously detecting the concentration of human serum albumin by using a UV/visible spectrophotometer. Concentration was stopped until the human serum albumin concentration reached 25 g/500 ml. Collecting concentrated solution, and placing in a refrigerator at 2-8deg.C for use.

Example 4 preparation of Compound Albumin resuscitation fluid D

In the raw materials adopted in the test, the human serum albumin from the gene recombination source is purchased from Wuhan Gramineae, the packaging specification is 12.5 g/50 ml glass bottle, and the albumin concentration is 25%. Sodium chloride is purchased from the company of Simarouba medical science and technology, and raw materials with package specifications are packaged. Potassium chloride was purchased from Guogou chemical Co., ltd, and was packaged to a GR specification. Anhydrous magnesium chloride was purchased from national pharmaceutical chemicals limited, package size 99.9% powder (wokak). Trisodium citrate dihydrate is purchased from national pharmaceutical group chemical reagent company, and has a package specification GR (Shanghai test) of 99.5% or more. Sodium acetate trihydrate is purchased from national pharmaceutical group chemical reagent company, and the packaging specification AR (Shanghai test) is more than or equal to 99.0%. Calcium gluconate is purchased from national pharmaceutical group chemical reagent company, inc., package size 99% (Woki). Anhydrous glucose is purchased from Shanxi jin ocean pharmaceutical excipients, inc., and is packaged into pharmaceutical grade. The water for injection is purchased from Shandong Mimo pharmaceutical Co., ltd, and the water for injection is sterilized in package specification.

The specific formula is as follows:

And a formula IV: compound albumin resuscitator D

Each 500ml contains:

gene recombination source human serum albumin 25 g

Sodium octoate 1.67g

Sodium chloride 1.516g

Potassium chloride 0.150g

Magnesium chloride 0.102g

Sodium citrate 0.294g

1.702G of sodium acetate

Calcium gluconate 0.336g

Anhydrous dextrose 5.001g

500Ml of water for injection

Because the human serum albumin injection from the gene recombination source contains small molecules such as salt ions, the test needs to be carried out by replacing the small molecules with a salt solution in a formula IV. The present invention employs a 30Kda ultrafiltration membrane pack (model number Biomax 30Kda, manufactured by Millipore Co.). A30 Kda ultrafiltration membrane pack, according to manufacturer's instructions, entraps compounds having a molecular weight greater than 30Kda, while compounds having a molecular weight less than 30Kda are filtered out. Therefore, the method can not only keep the human serum albumin (the molecular weight is 65-68 Kda), but also filter out small molecules such as salt ions in the human serum albumin injection. In addition, the 30Kda ultrafiltration membrane bag also has a concentration function, and can be concentrated to 1/5-1/20 of the original volume at most, so that the concentration of protein is improved.

The operation was performed according to the product operation manual provided by Millipore company of the manufacturer. The Biomax30Kda ultrafiltration membrane bag was properly mounted. The ultrafiltration membrane pack was placed on ice for pre-cooling for 30 minutes. The liquid peristaltic pump was turned on and the membrane pack was rinsed with sterile water for injection for 3 pack volumes. Then, 5 membrane package volumes were equilibrated with albumin-free compound albumin resuscitation fluid D. Adding the gene recombinant source human serum albumin injection into albumin-free compound albumin resuscitator D, uniformly mixing, replacing 5 membrane package volumes, concentrating, and simultaneously detecting the concentration of human serum albumin by using a UV/visible spectrophotometer. Concentration was stopped until the human serum albumin concentration reached 25 g/500 ml. Collecting concentrated solution, and placing in a refrigerator at 2-8deg.C for use.

Example 5 stability test of formulation

The above formulations one to four were each sterile filtered and filled into 50ml glass bottles, and stability at 57 degrees celsius was examined as compared with plasma-derived albumin and gene recombination-derived albumin. The judgment standard is as follows: and (5) precipitation occurs in the heated solution, so that the solution is thermolabile, and the experiment is ended. The results are shown below:

therefore, the formula of the invention has obviously enhanced thermal stability. This is feasible for the relief of traumatic hemorrhagic shock in extreme conditions such as field or natural disasters.

Example 6 bacterial infection

In clinical, the first 6 hours are critical periods in the resuscitation treatment of shock, so that liquid and needle are continuously replaced for wounded in the infusion treatment process, bacterial pollution is easily caused, and the wounded is infected. The experiment adopts the detection of the content of C-sensitive protein of shock rabbits to judge the condition of bacteria infection of the rabbits. The experimental results are as follows:

therefore, frequent liquid exchange obviously improves the content of C-sensitive protein, and can possibly cause bacterial infection of shock patients. In general, frequent clinical fluid changes have a low probability of causing infection. However, in shock patients, the body immunity is low due to blood loss, thereby increasing the probability of bacteria infection during infusion.

Example 7 comparative experiments of lyophilized plasma with the formulation of the invention

In antishock resuscitation fluid, lyophilized plasma and human serum albumin play a vital role. The freeze-dried blood plasma can greatly increase the survival rate of wounded by correcting clotting factor disturbance in war wound hemorrhagic shock. However, compared with the formula of the invention, the freeze-dried blood plasma is difficult to dissolve in water, is easy to generate sediment, has to be filtered, and is not resistant to high temperature. The experimental results are as follows:

Therefore, the freeze-dried blood plasma is precipitated due to intolerance to high temperature, and the rescue efficiency and safety of wounded are directly affected under extreme other conditions such as the wild or disasters.

Example 8 creation of shock animal model

1. Hemorrhagic shock animal model grouping:

The invention is exemplified by, but not limited to, hemorrhagic shock, which encompasses shock caused by burns, war wounds, infantile sepsis, chest blood, geological disasters, etc., and falls within the scope of the invention.

Healthy New Zealand rabbits, male, 72. Average body weight 2.5.+ -. 0.4kg. Supplied by the laboratory animal center in Changping parks in beijing. The cells were randomly divided into 12 groups of 6 cells.

The experimental group of the experimental animals:

2. The method for establishing the hemorrhagic shock animal model comprises the following steps:

The establishment of a model of hemorrhagic shock in rabbits, reference (Su Rong et al, effect of infusion temperature on liver function in hemorrhagic shock model rabbits. Southern Protect, 2004, 11 (6): 15-17) and literature (Liu Liangming: new concept for fluid resuscitation for war wound shock. National medical trauma & surgical basic problem handbook, 1998, 19 (2): 68-70).

The indwelling needle at the vein of the rabbit ear edge is connected with an intravenous infusion set controlled by an infusion pump, and the channel is used for intravenous infusion. The left external jugular vein was isolated and a Swan-Ganz floating catheter was inserted 5cm to collect venous blood samples. An arterial catheter was placed along the left femoral artery and connected to an HP Model 64s (M1166A) multifunctional monitor. The hemorrhagic shock model was prepared by bleeding through femoral artery cannulation for about 20 minutes, reducing Mean Arterial Pressure (MAP) to 5.33Kpa (40 mmHg), and maintaining rabbit MAP stable for about 30 minutes. The infusion rate of the antishock resuscitation fluid was controlled to 1.5 ml/(min. Kg) by the infusion pump.

The liquid resuscitation treatment scheme is as follows: after successful modeling of the rabbit model, mean Arterial Pressure (MAP) was reduced to 5.33Kpa (40 mmHg) and fluid resuscitation was initiated after maintaining MAP stable for about 30 minutes. The resuscitation fluid was infused only within 24 hours after injury, with the infusion time points being: 0.3, 9 and 18 hours. When MAP >80mmHg is detected, resuscitation fluid infusion is stopped.

The detection scheme is as follows: MAP detection time point: 0.6, 12 and 24 hours. Lac detection time point: 0.6, 12 and 24 hours. Survival rate detection time points: 0.1, 3 and 6 days.

The invention discloses a post-injury treatment and detection scheme:

example 9 treatment of hemorrhagic shock with different antishock resuscitation fluids

In examples 1-4, different formulations of anti-shock resuscitation fluid were used. From formulation one to formulation four, all served as experimental groups. The control group used a three-step combination (ringer's solution + albumin + saline) as the antishock resuscitation fluid. Meanwhile, a negative control group (shock recovery treatment with water for injection only) and a positive control group (normal health, without hemorrhagic shock treatment) are also provided.

After modeling, the therapeutic effect against shock resuscitation fluid was examined. The detection indexes adopted by the invention are as follows:

1. mean Arterial Pressure (MAP): the level of MAP is an important life recovery indicator. If the MAP is less than 5.33Kpa (40 mmHg), the animal is in shock state, and if the MAP is greater than 50mmHg, the animal may be resuscitated. When MAP is more than 80mmHg, the experimental animal is basically recovered, and the death rate is greatly reduced. It can be seen that the detection of MAP values is of great clinical significance and reference value.

2. Lactic acid (Lac): lactate index represents whether compensatory shock and systemic tissue oxygenation disorder are corrected, and only on the basis of hemodynamic index recovery, oxygen debt is repared, intra-tissue acidosis is corrected, and recovery from shock is complete upon recovery from aerobic metabolism.

3. Survival rate: the first 4-6 hours after hemorrhagic shock are critical phases of fluid resuscitation therapy, which determine the critical phase of animal survival. Therefore, the survival rate is the most representative, and the treatment effect of the anti-shock resuscitation fluid is fully reflected.

EXAMPLE 10 evaluation of the effectiveness of the anti-shock resuscitation fluid of the invention

The invention adopts different antishock resuscitation fluid formulas to carry out resuscitation treatment on hemorrhagic shock, and the antishock resuscitation fluid formulas of the invention are verified to have curative effect in treating shock by detecting various indexes of rabbit survival rate, MAP and Lac.

TABLE 1 MAP changes at various time points after shock for each group (mmHg)

As can be seen from the experimental data in Table 1 (MAP), the compound albumin resuscitation fluid (from formulation one to formulation four) of the present invention was subjected to antishock treatment, and MAP from 40mmHg to 80mmHg or more was raised in rabbits only for 6 hours, and MAP values were returned to positive control levels for 24 hours. MAP is the most important criterion for clinically judging the shock state and the resuscitation state of a wounded person. It is generally clinically recognized that a victim is in shock when MAP is less than 40 mmHg. When MAP >80mmHg, it is basically judged that the victim has been resuscitated. However, only when the MAP value reaches the level of a normal healthy person, it is possible to judge that the wounded person is completely resuscitated from the shock state to the awake state.

The three-step combination of the same group of experiments (sodium lactate ringer's solution + albumin + physiological saline) indicated sequential infusion of different resuscitation fluids. From experimental results, it took 24 hours from the onset of shock (MAP 40 mmHg) to resuscitative (MAP >80 mmHg) in the injured person.

From the aspect of the infusion times of the resuscitation fluid, the compound albumin resuscitation fluid is infused for 2 times, the wounded can be resuscitated after 6 hours (MAP >80 mmHg), and the three steps of combination (sodium lactate ringer's fluid, albumin and normal saline) need to be infused with the resuscitation fluid for 4 times (MAP >80 mmHg) in sequence to resuscitate the wounded.

Therefore, compared with the traditional clinically commonly used anti-shock resuscitation fluid (such as ringer's solution, albumin, physiological saline and the like), the compound albumin resuscitation fluid has more remarkable curative effect and better effect in the aspect of treating shock.

Lac index indicates whether acidosis is corrected in the injured tissue, and recovery from shock is complete only when aerobic metabolism is restored. Lac often accumulates rapidly in the body when the body's oxygen content is insufficient, and if it cannot be cleared rapidly, it causes the occurrence of acidosis in the wounded. Thus, lac detection data reflects the actual resuscitation of the injured person after shock and reflects the subsequent complications. Referring to the test data of table 2 (Lac), it is known that the rabbit is in shock state, a large amount of lactic acid is accumulated in vivo, and the accumulation of lactic acid is rapidly increased with the lapse of time, and the rabbit is dead after a certain amount of lactic acid is accumulated. Thus, when hemorrhagic shock occurs, rapid anti-shock therapy is the key point, and the most effective route is to infuse large amounts of anti-shock resuscitation fluid.

TABLE 2 Lac Change (mmol/L) at various time points after shock for each group

From the experimental results in table 2, the compound albumin resuscitation fluid was superior to the three-step combination (sodium lactate ringer's solution + albumin + physiological saline), and the Lac content in the blood was reduced to the positive control level (normal health) only by 6 hours. But the three-step combination group (sodium lactate ringer's solution + albumin + physiological saline) took 24 hours to reduce Lac to the level of positive control (normal health). Therefore, compared with the clinically commonly used anti-shock resuscitation fluid (such as ringer's solution, albumin, physiological saline and the like), the compound albumin resuscitation fluid can greatly shorten the concentration of Lac in the body, which is really calculated as the complete shock resuscitation.

Table 3, change in survival at various time points after shock for each group

Remarks: in the table "; "×" indicates death.

Table 3 statistics of survival rates of different antishock resuscitation fluids during treatment of rabbit shock.

The calculation formula of the survival rate is as follows: survival = (number of survivors/total number) x 100%.

From table 3 (survival rate), the survival rate of the compound albumin resuscitation solution was 100% within 6 days. In comparison, the survival rate of the three-step combination (sodium lactate ringer's solution + albumin + physiological saline) was 54% over 6 days. It can be seen that the compound albumin resuscitation fluid is superior to the clinically commonly used anti-shock resuscitation fluid (e.g. ringer's solution, albumin, physiological saline, etc.) in terms of anti-shock treatment.

From the detection indexes of MAP, lac and survival rate, the compound albumin resuscitation fluid is superior to the clinically commonly used anti-shock resuscitation fluid (such as ringer's solution, albumin, physiological saline and the like) in terms of the anti-shock resuscitation fluid, and has obvious difference in treatment effect. Therefore, the compound albumin resuscitation fluid has definite and reliable curative effect in the aspect of shock treatment and has good clinical popularization value.

The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains.

Claims

1. An antishock resuscitation fluid comprising the following composition: albumin, sodium octoate and crystal fluids.

2. The antishock resuscitation fluid of claim 1, wherein: each 500ml of the anti-shock recovery liquid contains 2.5-50 g of albumin, 1.67 g of sodium octoate, 3.00-10 g of salt in the crystal liquid and 500ml of water for injection.

3. The antishock resuscitation fluid of claim 1 or 2, characterized in that: the albumin is at least one of blood plasma source human albumin and gene recombination source human albumin;

the crystal liquid is any one of sodium chloride physiological saline and ringer's solution.

4. The anti-shock resuscitation fluid according to any one of claims 1-3, wherein: the crystal liquid is selected from the group consisting of: sodium lactate ringer's solution and sodium acetate ringer's solution.

5. The anti-shock resuscitation fluid according to any one of claims 1-4, wherein: the antishock resuscitation fluid consists of the following formula:

Each 500ml contains:

2.5-50 g of plasma source human serum albumin;

sodium octoate 1.67g

Sodium lactate 1.55g

Sodium chloride 1.33g

Potassium chloride 0.15g

Calcium chloride dihydrate 0.10g

500Ml of water for injection.

6. The anti-shock resuscitation fluid according to any one of claims 1-4, wherein: the antishock resuscitation fluid consists of the following formula:

Each 500ml contains:

2.5-50 g of plasma source human serum albumin;

sodium octoate 1.67g

Sodium chloride 1.516g

Potassium chloride 0.150g

Magnesium chloride 0.102g

Sodium citrate 0.294g

1.702G of sodium acetate

Calcium gluconate 0.336g

Anhydrous dextrose 5.001g

500Ml of water for injection.

7. The anti-shock resuscitation fluid according to any one of claims 1-4, wherein: the antishock resuscitation fluid consists of the following formula:

Each 500ml contains:

2.5-50 g of gene recombination source human serum albumin;

sodium octoate 1.67g

Sodium lactate 1.55g

Sodium chloride 1.33g

Potassium chloride 0.15g

Calcium chloride dihydrate 0.10g

500Ml of water for injection.

8. The anti-shock resuscitation fluid according to any one of claims 1-4, wherein: the antishock resuscitation fluid consists of the following formula:

Each 500ml contains:

2.5-50 g of gene recombination source human serum albumin;

sodium octoate 1.67g

Sodium chloride 1.516g

Potassium chloride 0.150g

Magnesium chloride 0.102g

Sodium citrate 0.294g

1.702G of sodium acetate

Calcium gluconate 0.336g

Anhydrous dextrose 5.001g

500Ml of water for injection.