CN1631876A - Preparation of Probenecid Sodium and Potassium and Combination of Probenecid Sodium and Potassium with β-Lactam Antibiotics and Its Application - Google Patents

Abstract

The invention relates to a compound injection composed of probenecid sodium and potassium and beta-lactam antibiotics and application thereof, belonging to the field of chemical pharmacy. The probenecid is prepared into sodium salt and potassium salt, and then is prepared into powder injection and freeze-dried powder injection together with the beta-lactam antibiotic, and the powder injection and the beta-lactam antibiotic are simultaneously injected into the vein or infused into the vein. Can obviously prolong the blood elimination half-life of antibiotics_1/2β) Increasing the area under the curve (AUC), and obviously prolonging the time that the blood concentration of the beta-lactam antibiotics in the compound preparation exceeds the MIC of the corresponding bacteria. Reducing abuse of antibiotics, avoiding occurrence and development of drug-resistant strains, having good preparation safety, definite curative effect, stable quality and more convenient use.

Description

Preparation of Probenecid Sodium and Potassium and Combination of Probenecid Sodium and Potassium with β-Lactam Antibiotics and Its Application

Technical field:

The invention relates to the preparation of probenecid sodium and potassium and the composition of a compound preparation with β-lactam antibiotics and its application, including powder injection and freeze-dried powder injection, belonging to the field of chemical pharmacy.

Background technique:

Probenecid can competitively inhibit the secretion of many β-lactam antibiotics in the renal tubules, prolong their blood elimination half-life (t _1/2β ), increase the area under the curve (AUC), and increase the blood drug concentration Level, so that their blood concentration exceeds the minimum inhibitory concentration (MIC) of the corresponding bacteria significantly prolongs, thereby increasing their curative effect. At present, there is no compound prescription made of probenecid and β-lactam antibiotics, but only one oral compound preparation composed of probenecid and ampicillin, that is, ampicillin probenecid capsules. In addition, since probenecid itself is not soluble in water but only soluble in organic solvents, it cannot form compound prescriptions with β-lactam antibiotics for injection. We have confirmed through a large number of experiments that probenecid sodium (potassium) not only can be dissolved in water, but also has the same pharmacological effect as probenecid. At present, there is no report on probenecid sodium (potassium) itself and compound preparations composed of antibiotics. Probenecid sodium (potassium) and β-lactam antibiotics form a compound prescription, which can reduce the amount of β-lactam antibiotics in the compound prescription, thereby effectively reducing the occurrence and occurrence of bacterial resistance caused by ultra-high doses of antibiotics. Development; also can save the resource of antibiotic, reduce the misuse of antibiotic; Because the probenecid sodium (potassium) in the compound can prolong the blood elimination half-life of the antibiotic in the compound, this just makes the dosing interval prolong, convenient for the patient's use.

Chemical structural formula of Probenecid

Invention content:

The object of the present invention is to provide a kind of probenecid sodium, potassium and preparation thereof, probenecid is made into the probenecid sodium (potassium) raw material powder meeting the human medicine injection standard by changing the process approach, and the content of probenecid is changed. The physical and chemical properties make probenecid sodium (potassium) soluble in water.

Another object of the present invention is to provide a compound prescription composed of probenecid sodium, probenecid potassium and β-lactam antibiotics and its application. Probenecid sodium (potassium) and β-lactam antibiotics include penicillins Antibiotics and cephalosporin antibiotics consist of powder injections and freeze-dried powder injections, which are intravenously injected or intravenously dripped together with β-lactam antibiotics. It can effectively prolong the half-life (t _1/2 ) of β-lactam antibiotics, increase the area under the curve (AUC), and eliminate the hemolytic property of probenecid sodium (potassium) The safety in the human body plays a protective role, and can safely and effectively enter the human body through the intravenous route as a new compound preparation. The compound medicine composed of probenecid sodium (potassium) and β-lactam antibiotics, because it directly enters the blood circulation, can be completely absorbed by the human body, can quickly reach the peak, and has a rapid effect and more significant curative effect.

Technical scheme of the present invention is realized like this:

1. (1) Probenecid sodium: p-[(dipropylamino)sulfonyl]sodium benzoate

The structural formula is:

Molecular formula: C ₁₃ H ₁₈ NO ₄ SNa Molecular weight: 307.34

(2) Probenecid Potassium: Potassium p-[(dipropylamino)sulfonyl]benzoate

The structural formula is:

Molecular formula: C ₁₃ H ₁₈ NO ₄ K Molecular weight: 323.45

2, (1) The preparation technology of sodium sulfonidate is completed in the following steps (by weight):

First, dissolve 0.9-102 sodium hydroxide in 11.5-1308 purified water (or water for injection), add 6.5-663 probenecid, heat to 60±10°C, adjust the pH to 7.5-10, and keep stirring until the pH The value is basically constant; under the condition of 60±10°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions. The raw material of sodium sulfonidate is white or off-white crystalline powder; calculated as dry product, the content of probenecid should not be less than 88.50%.

Its chemical reaction formula is:

(2) the preparation technology of probenecid potassium is finished (by weight) as follows::

First, dissolve 0.17-173.44 potassium hydroxide in 11.5-1308 purified water (or water for injection), add 0.88-885.75 probenecid, heat to 60±10°C, adjust the pH to 7.5-10, and keep stirring until the pH The value is basically constant; under the condition of 60±10°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions. The raw material of sodium sulfonidate is white or off-white crystalline powder; calculated as dry product, the content of probenecid should not be less than 86.25%.

The chemical reaction formula is:

3. A compound preparation composed of probenecid sodium (potassium) and β-lactam antibiotics, probenecid made into sodium (calculated as dry basis of probenecid) or potassium of probenecid (calculated as dry basis of probenecid) It is mixed with β-lactam antibiotics (based on the dry basis of each antibiotic) at a ratio of 1:1~3, mixed by conventional techniques, and subpackaged to make powder injections; β-lactam antibiotics include penicillin antibiotics and cephalosporins steroid antibiotics;

Penicillin antibiotics include: penicillin G and ampicillin.

Cephalosporin antibiotics include: cefazolin, cefuroxime, cefotaxime, and cefoxitin.

4. Probenecid sodium (potassium) combined with β-lactam antibiotics including penicillins and cephalosporins can effectively prolong the half-life (t _1/2 ) of β-lactam antibiotics and increase the area under the curve (AUC), while also eliminating the hemolytic properties of probenecid sodium (potassium).

In order to better understand the essence of the present invention, the new application of probenecid sodium (potassium) and the above-mentioned β-lactam antibiotics to form a compound preparation will be explained below with our test results. Related experimental research on ampicillin sodium/probenecid sodium

1. Ampicillin Sodium/Probenecid Sodium Acute Toxicity Test

1. Summary

This experiment adopts mouse single intravenous and intraperitoneal injection routes to observe the acute toxic reaction of ampicillin sodium/probenecid sodium to mice, and the LD ₅₀ and 95% credible limits of intravenous injection of mice are measured as 2594mg/kg and (2380-2858) mg/kg; the measured LD ₅₀ and 95% confidence limits of intraperitoneal injection mice were 3113mg/kg and (2880-3383) mg/kg, respectively.

2. Purpose

After a single intravenous and intraperitoneal injection of ampicillin sodium/probenecid sodium to mice, observe the toxic reaction and determine the LD ₅₀ , to provide acute toxicity test data for clinical application.

3. Test materials

(1) Drugs: ampicillin sodium/probenecid sodium (3:1), dissolved in normal saline before the test and used

(2) Animals: ICR clean level mice, weighing 18-20 g, provided by the Experimental Animal Center of Jilin University School of Basic Medicine, certificate number: SCXK (Ji) 2003-0001.

4. Methods and results

(1) Determination of LD ₅₀ of intravenous injection of ampicillin sodium/probenecid sodium in mice

According to the pre-test results, the group spacing was calculated to be 1:0.85, and they were divided into 5 dose groups. Take 50 mice, and divide them into 5 groups at random, 10 in each group, half male and half male, inject intravenously with 0.15ml/10g body weight respectively, observe and record the animal reaction symptoms and death time immediately after administration, observe continuously for 14 days, the results It was shown that after the administration of the high dose group, the mice convulsed and died, and the activities of the other dose groups decreased after administration. The dead mice were immediately dissected, and no abnormal changes in various organs were seen with the naked eye. With the reduction of the drug dose, the mice died. According to the mortality rate of each dose group, the LD ₅₀ and 95% confidence interval were calculated by SPSS processing software, and the results are shown in Table 1.

Table 1 Acute Toxicity Experiment Results of Ampicillin Sodium/Probenecid Sodium Intravenous Injection in Mice

Dose Log Dose Number of Animals Number of Deaths Mortality LD50 and 95% Confidence Interval

mg/kg (X) (only) (only) (%)

3375 3.53 10 10 100.00 2594mg/kg,

2868 3.46 10 6 60.00 (2380～

2438 3.39 10 3 3 30.00 2858)mg/kg

2072 3.32 10 2 20.00

1761 3.25 10 0 0 00.00

(2) Determination of LD ₅₀ of intraperitoneal injection of ampicillin sodium/probenecid sodium in mice

According to the pre-test results, the group spacing was calculated to be 1:0.85, and they were divided into 5 dose groups. Take 50 mice and divide them into 5 groups at random, 10 in each group, half male and half male, inject intraperitoneally with 0.18ml/10g body weight respectively, observe and record the animal reaction symptoms and death time immediately after administration, observe continuously for 14 days, the results It was shown that after the administration of the high dose group, the mice convulsed and died, and the activities of the other dose groups decreased after administration. The dead mice were immediately dissected, and no abnormal changes in various organs were seen with the naked eye. With the reduction of the drug dose, the mice died. According to the mortality rate of each dose group, the LD ₅₀ and 95% confidence interval were calculated by SPSS processing software, and the results are shown in Table 2.

Table 2 Acute toxicity test results of ampicillin sodium/probenecid sodium intraperitoneal injection in mice

Dose Log dose Number of animals Deaths Mortality LD ₅₀ and 95% CI

mg/kg (X) (only) (only) (%)

4050 3.61 10 10 100.00 3113mg/kg,

3442 3.54 10 7 70.00 (2880～

2926 3.47 10 3 30.00 3383)mg/kg

2487 3.40 10 1 10.00

2114 3.33 10 0 0 00.00

5 Conclusion

Using a single intravenous and intraperitoneal injection of ampicillin sodium/probenecid sodium in mice, the LD ₅₀ and 95% confidence limits of intravenous injection of mice were measured to be 2594 mg/kg and (2380～2858) mg/kg respectively; The LD ₅₀ and 95% confidence limits of intraperitoneal injection of mice were 3113 mg/kg and (2880-3383) mg/kg, respectively.

2. Subacute toxicity test of ampicillin sodium/probenecid sodium in rats

1. Abstract In this experiment, two doses of ampicillin sodium/probenecid sodium were injected intraperitoneally into rats. The doses of ampicillin sodium/probenecid sodium were: high dose 0.9g/kg; low dose 0.4g/kg; (equivalent to 1/2.5, 1/5.5 of rat LD ₅₀ ). The volume of intraperitoneal injection was 0.5ml/100g body weight for three consecutive weeks. The results of the subacute toxicity test showed that ampicillin sodium/probenecid sodium had no significant effect on animal food intake, appearance behavior, body weight, hair growth, and excretion of feces; Compared with the control group, there was no significant difference in visceral coefficient; there was no significant difference in blood routine compared with the control group; the blood urea nitrogen (UREA) high-dose group had a slight increase in blood biochemical indicators, and there was a difference compared with the control group (P<0.05 , Other indicators were not significantly different from those in the control group, and the morphological changes of various organs in histopathological examination were not significantly different from those in the control group. It is suggested that long-term medication may cause a transient increase in blood urea nitrogen.

2. The purpose of the test To observe and determine whether rats have toxic reactions due to drug accumulation after long-term intraperitoneal injection of ampicillin sodium/probenecid sodium, and provide the target organs of toxic reactions and the reversible degree of damage, which is the safety of clinical medication. provide a test basis.

3. Test materials

(1) Drug: ampicillin sodium/probenecid sodium (3:1), dissolved in normal saline before use

(2) Animals: Wistar rats, weighing 80-100 g, half male and half male, provided by the Animal Center of Jilin University School of Basic Medicine. Quality certificate number: SCXK (Ji) 2003-0001.

4. Test method

Wistar rats aged 6-8 weeks (body weight 80-100g) were used in the experiment. After the animals were fed normally for one week, they were randomly divided into three groups according to their body weight, with 5 male and female rats in each group, administered by intraperitoneal injection at 9:00 am every day, ampicillin sodium The dose of probenecid sodium is: high dose 0.9g/kg; low dose 0.4g/kg; administration volume is 0.5ml/100g body weight, and the control group is given the same volume of normal saline. During the dosing period, observe the general state of the animals, behavioral activities, diet and urine and feces, record abnormal changes at any time, weigh once a week, record food intake every day, and recalculate the dosing amount according to the change in body weight. Five animals in each group were taken from the abdominal aorta to measure hematology and blood biochemical indicators, including red blood cell count, white blood cell count, platelet count, white blood cell classification and hemoglobin determination; the serum was separated for liver and kidney function tests, and then taken out And weigh rat organs, including heart, liver, spleen, lung, kidney, adrenal gland, pancreas, stomach, duodenum, colon, rectum, thyroid, thymus, brain, pituitary, eyeball, optic nerve, spinal cord, medulla oblongata, bladder , testis (epididymis), uterus (ovary), prostate, manubrium sternum, trachea, lymph nodes, calculate the coefficient of major organs according to body weight, and carry out gross and microscopic pathological examination on each dose group, and the remaining animals continue to be raised after stopping the drug, and observe The recovery period varies.

5. Test results

(1) General observation:

Ampicillin sodium/probenecid sodium was administered to rats by intraperitoneal injection of high and low doses for three consecutive weeks, and all the tested animals survived. Before administration, during administration and recovery period, the animals in each group had normal activities, lively behavior and normal growth. , with a smooth coat, normal diet, normal urine and stool, and no other abnormal signs. The test results showed that continuous intraperitoneal injection of ampicillin sodium/probenecid sodium had no significant effect on the behavioral activities and appearance signs of the rats.

(2) Effects on body weight and food intake

Rats in each dosage group had normal diet and drinking water before administration, during administration and recovery period, and their body weight increased, and there was no significant difference between the administration group and the control group (see Table 1.1, 1.2, 2.1).

Table 1.1 The impact of ampicillin sodium/probenecid sodium subacute toxicity test on food intake during administration of rats (g) (x ± s)

after administration

Group Before administration

|

Blank group 15.1±1.42 17±2.24 17.0±2.86 18.2±1.60

High-dose group 15.9±1.36 16.7±2.76 17.8±1.99 18.8±1.33

Low-dose group 14.7±1.24 16.9±3.68 17.1±3.21 18.1±1.76

Compared with the control group, P＞0.05

Table 1.2 Effect of Ampicillin Sodium/Probenecid Sodium Subacute Toxicity Test on Food Consumption of Convalescent Rats (g) (x±s)

Group 1W 2W

Blank group 20.97±1.04 22.77±1.10

High dose group 20.17±1.13 22.46±1.68

Low dose group 20.83±1.13 22.00±1.63

Compared with the control group, P＞0.05

Table 2.1 Effect of Ampicillin Sodium/Probenecid Sodium Subacute Toxicity Test on Rat Body Weight Gain (g) (x±s)

Groups Control group 0.9g/kg high dose group 0.4g/kg low dose group

Before administration 112.3±7.009 113.3±5.982 107.3±3.974

1W 151.6±12.05 143.7±19.18 140.4±15.39

2W 173.5±16.83 167.3±18.98 174.7±21.64

3W 210±23.68 197.1±26.78 196.4±23.65

recovery period

1W 224±27.55 226±18.87 225.2±41.07

2W 247±17.77 248±22.97 249.8±20.41

Compared with the control group, P＞0.05

6. Clinical examination:

(1) Hematology:

During the administration period, the white blood cells (WBC), red blood cells (RBC), lymphocytes (LY), hemoglobin (Hb), platelets (PLT) and neutral lobes (GRAN) were measured by the American Abbott blood cell counter in each dosage group.

Test results: the hematological indexes of the animals in the administration group were all within the normal range, and compared with the control group, there was no significant difference (see Table 3)

Table 3 The effect of ampicillin sodium/probenecid sodium on the hematological indicators of rats during the administration period (n=5 x ± s)

Grouping Units Control group 0.9g/kg high dose group 0.4g/kg low dose group

WBC ×10 ⁹ /L 6.98±1.30 7.36±2.65 6.7±2.22

LY% 79.86±3.49 78.6±5.13 78.68±4.90

Plt ×10 ⁹ /L 710.6±103.67 661.8±117.29 664±123.77

GR % 12.4±3.48 14.06±6.21 13.96±5.00

RBC ×10 ¹² /L 6.98±0.30 6.71±0.18 6.99±0.26

HB (g/L) 136.4±5.13 134±5.43 136.4±3.04

Compared with the control group, P＞0.05

(2) Blood biochemistry:

During the administration period, each dosage group was measured with a Hitachi (7150) automatic biochemical analyzer for aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), urea nitrogen (UREA ), total protein (TP), albumin (ALB), blood glucose (GLU), total bilirubin (TBIL), creatinine liver (CRE), and total cholesterol (CHO).

Test results: the blood urea nitrogen (UREA) of the ampicillin sodium/probenecid sodium administration high-dose group was slightly elevated, which was different from that of the control group (P<0.05=, other biochemical indicators were all within the normal range, compared with the control group There was no significant difference between groups (see Table 4).

Table 4 Effect of Ampicillin Sodium/Probenecid Sodium on Blood Biochemical Indexes of Rats (n=5 x±s)

Grouping Units Control group 0.9g/kg high dose group 0.4g/kg low dose group

ALT U/L 73.46±17.87 69.58±13.78 64.38±9.44

TP g/L 63.26±3.46 63.18±2.90 64.78±2.33

ALB g/L 30.58±2.06 29.9±2.28 31.28±2.11

ALP U/L 251±38.63 266.5±31.37 261.36±2.65

AST U/L 232.82±8.70 235.72±34.77 223.18±9.07

UREA mmol/L 8.48±0.26 10.052±1.22 ^* 9.698±0.76

CR μmol/L 71.66±4.97 80.96±13.72 75.94±8.54

CHO mmol/L 2.146±0.23 2.184±0.28 2.174±0.26

GLU mmol/L 6.43±0.62 6.378±0.63 6.564±0.614

T-BIL μmol/L 7.54±0.56 7.42±0.63 7.36±0.50

Compared with the control group: ^* p<0.05

7. Histological examination

(1) Gross observation: Rats were sacrificed after intraperitoneal injection of ampicillin sodium/probenecid sodium for three weeks, and the main organs of each dose group were visually observed (including heart, liver, spleen, lung, kidney, stomach, adrenal gland, thyroid gland, Brain, thymus, testis (epididymis), prostate, uterus (ovary) had no significant changes, and organ coefficients had no significant difference compared with the control group (see Table 5).

Table 5 Ampicillin Sodium/Probenecid Sodium is on the influence (x±sn=5g/100g) of administration period rat organ coefficient

Groups Control group High dose group Low dose group

Dose 0.9g/kg 0.4g/kg

Heart 0.4957±0.0894 0.3985±0.0733 0.4813±0.0761

Liver 4.6767±1.0663 3.9651±0.5274 4.8682±1.2375

Spleen 0.2307±0.0446 0.2106±0.0276 0.2301±0.0392

Lung 0.7609±0.1448 0.6713±0.1052 0.7300±0.1329

Kidney 0.9776±0.1883 0.8948±0.2878 0.9661±0.2180

Adrenal gland 0.0190±0.0098 0.0122±0.0064 0.018±0.0110

Thyroid 0.0076±0.0039 0.0071±0.0025 0.0112±0.0064

Brain 0.6864±0.1201 0.6438±0.1116 0.6926±0.1146

Thymus 0.3537±0.0547 0.3148±0.0729 0.3778±0.1226

Testis (epididymis) 1.3072 1.3562 1.2837

Prostate 0.1379 0.1091 0.1629

Uterus (ovary) 0.2557 0.1989 0.3036

Compared with the control group, P＞0.05

(2) Observation under the microscope: three weeks after intraperitoneal injection of ampicillin sodium/probenecid sodium in rats, the specimens of each major organ (same as above) in each dose group and control group were routinely fixed, HE stained, histologically sliced, and microscopically Under observation, no obvious pathological changes were seen, and the results were shown in the pathology report.

8. Test conclusion

Three weeks after intraperitoneal injection of ampicillin sodium/probenecid sodium high-dose and low-dose groups of rats, there was a difference (P<0.05) between the high-dose blood urea nitrogen (UREA) group and the control group in blood biochemical indicators (P<0.05), suggesting that: Long-term medication should monitor changes in renal function, other indicators were not significantly different from those in the control group, and the morphological changes of various organs in histopathological examination were not significantly different from those in the control group.

3. Vascular irritation test of ampicillin sodium/probenecid sodium

1. Abstract: The test shows that the intravenous infusion of ampicillin sodium/probenecid sodium has no obvious vascular irritation.

2. Purpose of the test: To observe whether ampicillin sodium/probenecid sodium has vascular irritation through the test.

3. Test drug: ampicillin sodium/probenecid sodium (3:1), dissolved in normal saline for use during the test. 4. Test animals: New Zealand big-eared white rabbits, weighing 2.3-2.5kg. Provided by the Experimental Animal Department of Jilin University. Certificate number: SCXK (Ji) 2003-0001.

5. Test method: Take 4 healthy rabbits, half male and half male. According to body weight and sex, they were divided into 0.9% sodium chloride injection control group and ampicillin sodium/probenecid sodium, with 2 rats in each group. Ampicillin sodium/probenecid sodium was slowly infused into the left ear vein of rabbits at 10ml/kg, once a day for 7 consecutive days. The control group was intravenously infused with 0.9% sodium chloride injection in the same way. In addition to observing the local manifestations of drug administration during and after each administration, the auricle on the side of the drug was cut off after the last intravenous infusion. cm wide specimens, a total of 3 specimens were taken. Sections were stained for pathological observation under a microscope.

6. Results: There was no dilatation of blood vessels in the auricle of the rabbits, no redness and swelling; pathological examination under the microscope showed no inflammatory cell infiltration in the control group and the drug group, and no obvious changes.

7. Conclusion: Ampicillin sodium/probenecid sodium was dripped 10ml/kg into rabbit's ear vein, once a day, for 7 consecutive days. The results of macroscopic and microscopic pathological examination showed that the intravenous infusion of ampicillin sodium/probenecid sodium had no local irritation.

4. Hemolysis test of ampicillin sodium/probenecid sodium

1. Abstract: The test shows that Ampicillin Sodium/Probenecid Sodium has no hemolysis and erythrocyte agglutination.

2. The purpose of the test: To observe whether ampicillin sodium/probenecid sodium has hemolysis and erythrocyte agglutination.

3. Test drugs: ampicillin sodium/probenecid sodium, provided by the entrusting party. It was used after dissolving with physiological saline during the test.

4. Test animals: New Zealand big-eared white rabbits, weighing 2.3-2.5kg. Provided by the Experimental Animal Department of Jilin University.

5. Test method: Take the blood of New Zealand big-eared white rabbits without fibrin, add normal saline to shake well, pour off the supernatant after centrifugation, measure and dilute red blood cells to 2% red blood cell suspension for test use. Take a number of test tubes, number them as shown in Table 1, add various test solutions, shake gently, put them in a 37°C water bath, observe and record the results for 0.5, 1, 2, 3, and 4 hours respectively, and observe whether there is any Hemolysis and erythrocyte agglutination, and placed at room temperature for 24 hours, continue to observe whether hemolysis and erythrocyte agglutination occur.

6. Test results: Ampicillin sodium/probenecid sodium was observed in a 37°C water bath for 4 hours. Except for the hemolysis control, the seventh tube showed obvious pink hemolysis and no red blood cell sedimentation. No hemolysis and erythrocyte agglutination occurred in 4h and 4h, and the erythrocytes were seen to sink. After shaking the test tube, the sinking erythrocytes floated up, and no erythrocyte agglutination was seen.

7. Conclusion: Ampicillin sodium/probenecid sodium was observed in a water bath at 37°C for 4 hours at the concentrations shown in Table 2 and Table 3, and no hemolysis was observed, and no hemolysis and agglutination were observed at room temperature for 24 hours.

Table 1 Operation sequence of ampicillin sodium/probenecid sodium hemolytic test Test tube number 1 2 3 4 5 6 7 Ampicillin sodium/probenecid sodium (ml) (see Table 2 and Table 3 for concentration) 0.1 0.2 0.3 0.4 0.5 - - Normal saline (ml) 2.4 2.3 2.2 2.1 2.0 2.5 - Distilled water (ml) - - - - - - 2.5 2% red blood cell suspension (ml) 2.5 2.5 2.5 2.5 2.5 2.5 2.5

Table 2 Ampicillin Sodium/Probenecid Sodium Hemolysis Test

10% mixed 22.5% mixed 45% mixed Probenecid 1.11% is equivalent to undissolved undissolved undissolved undissolved undissolved probenecid 2.2% is equivalent to propane Undissolved sulfonid 10%

Table 3 Ampicillin Sodium/Probenecid Sodium Hemolysis Test

5. Pharmacokinetic test of ampicillin sodium/probenecid sodium in beagle dogs

1. Purpose: This experiment aims to observe the pharmacokinetic process of ampicillin sodium after single intravenous injection of ampicillin and joint administration of ampicillin sodium/probenecid sodium in dogs at the same dose, so as to confirm the intravenous administration of probenecid sodium Whether the pathway can prolong the area under the curve (AUC) and blood elimination half-life (t _1/2β ) of ampicillin sodium will lay the foundation for the formulation modification of the compound preparation.

2. Test materials

2.1 Drugs and reagents

Ampicillin sodium and ampicillin sodium/probenecid sodium; Enalapril maleate (internal standard, purity: 99.8% provided by Beijing Institute for the Control of Pharmaceutical and Biological Products); supply).

Methanol was chromatographically pure and was purchased from Tianjin Concord Technology Co., Ltd. Other reagents were analytically pure and were products of Tianjin Fuchen Chemical Reagent Factory and Beijing Chemical Factory.

2.2 Test animals

Six adult Beagle dogs with a weight range of 10.0±1.6kg were purchased from the Experimental Animal Center of Sichuan Academy of Medical Sciences, certificate number: Yidongzi No. 24103123.

2.3 Experimental program and administration method

Six dogs were randomly divided into two groups (control group R and test group T) for a two-period crossover test, with the interval between two administration cycles being 7 days. The dosage and method of administration are as follows:

Control group: Each dog was given 40, 80 and 160 mg·kg ^-1 of ampicillin sodium reference substance alone, dissolved in normal saline, and then intravenously injected.

Experimental group: each dog was given ampicillin sodium/probenecid sodium 40/13.5, 80/27 and 160/54 mg·kg ^-1 , dissolved in normal saline, and then intravenously injected.

2.4 Sample collection and processing

Time points for intravenous blood collection: 0.17, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 10.0, 12.0h. (The control group continued to take 24.0, 36.0, 48.0h)

About 1 ml of blood was collected from the small saphenous vein of the dog's hind limb according to the above blood collection time points. Immediately after the blood sample was taken out, it was transferred into a heparin test tube, centrifuged (3500rpm) for 10min, the plasma was separated, and stored frozen in a -20°C refrigerator until assayed.

2.5 Plasma sample analysis method

Canine plasma samples were determined by LC/MS/MS method.

2.6 Data processing

List the blood drug concentration data after each dog intravenous injection of ampicillin sodium and ampicillin sodium//probenecid sodium, give the blood drug concentration-time curve, and according to the blood drug concentration-time data, use Topfit 2.0 software (Thomae GmbH, Germany)'s non-compartmental model to calculate the pharmacokinetic parameters of dogs after intravenous injection and oral administration, and compare the t _1/2β of ampicillin of the control group and the test group, AUC _0-t and AUC _{0 -∞} were compared to see if the results were statistically different.

3. Results and discussion

3.1 Plasma drug concentration and pharmacokinetic parameters after intravenous administration in Beagle dogs

6 Beagle dogs were intravenously injected low, middle and high three dosage groups of ampicillin sodium (control group) and ampicillin sodium/probenecid sodium (test group) after the ampicillin sodium average blood drug concentration-time curve as shown in Figure 1 shown. It can be seen from Figure 1 that after intravenous injection of ampicillin sodium/probenecid sodium, the average blood concentration of ampicillin at each blood collection time point was higher than that of the control group. The pharmacokinetic parameters of the control group and the test group are shown in Tables 1 to 6, respectively.

3.2 Results

Beagle dogs were given ampicillin sodium alone (control group), when the doses were 40, 80 and 160 mg·kg ^-1 , the t _1/2β of ampicillin sodium in plasma were 0.70±0.10, 0.92±0.07 and 1.49±0.13 h; AUC _0-t were 129.8±28.04, 142.8±24.92 and 233.7±48.71μg·h·mL ^-1 , AUC _0-∞ were 130.1±28.07, 143.0±24.94 and 234.0±48.78μg·h·mL ^{- 1} ; Beagle dogs were intravenously administered ampicillin sodium/probenecid sodium (test group), and the dose was 40/13.5, 80/27 and 160/54 mg·kg ^-1 respectively, the t _1/2β of ampicillin sodium in blood plasma 0.98±0.09, 1.15±0.11 and 1.75±0.21h; AUC _0-t were 160.2±16.27, 171.8±41.79 and 275.5±79.55μg·h·mL ^-1 , AUC _0-∞ were 160.5±16.29, 172.1±41.79 and 276.1±79.77 μg·h·mL ⁻¹ . It is particularly worth mentioning that the time for the blood concentration of ampicillin in the test group of the high, medium and low dose groups to exceed the minimum inhibitory concentration (MIC) of a specific bacterium was two hours longer than that of the control group.

The pharmacokinetics of table 1 low-dose control group ampicillin sodium Table 2 The pharmacokinetics of low-dose test group ampicillin sodium

Parameters Parameters Parameters

t _1/2 AUC _0-t AUC _0-∞ t _1/2 AUC _0-t AUC _0-∞

Low Dose-R Low Dose-T

(h) (μg·h·mL ^-1 ) (μg·h·mL ^-1 ) (h) (μg·h·mL ^-1 ) (μg·h·mL ^-1 )

1 0.80 127.5 128.1 1 0.98 150.3 150.4

2 0.81 184.0 184.2 2 0.99 186.2 186.4

3 0.64 121.9 122.1 3 0.89 148.5 148.8

4 0.58 118.8 118.8 4 0.95 160.7 161.1

5 0.63 101.9 102.0 5 0.94 171.8 172.3

6 0.74 124.8 125.1 6 1.15 143.6 143.8

Mean 0.70 129.8 130.1 Mean 0.98 160.2 160.5

SD 0.10 28.04 28.07 SD 0.09 16.27 16.29

Pharmacokinetic parameters of dosage group ampicillin sodium in table 3 Pharmacokinetic parameters of dosage group ampicillin sodium in table 4

t _1/2 AUC _0-t AUC _0-∞ t _1/2 AUC _0-t AUC _0-∞

Medium Dose-R Medium Dose-T

(h) (μg·h·mL ^-1 ) (μg·h·mL ^-1 ) (h) (μg·h·mL ^-1 ) (μg·h·mL ^-1 )

1 0.97 132.9 133.0 1 0.99 152.3 152.6

2 0.98 141.1 141.4 2 1.09 131.8 132.0

3 0.89 100.8 100.9 3 1.13 120.5 121.0

4 0.82 158.6 158.7 4 1.30 213.3 213.7

5 0.86 173.4 173.6 5 1.15 208.0 208.4

6 0.99 150.2 150.3 6 1.25 204.8 205.0

Mean 0.92 142.8 143.0 Mean 1.15 171.8 172.1

SD 0.07 24.92 24.94 SD 0.11 41.79 41.79

Table 3-11 Pharmacokinetic parameters of ampicillin in high-dose control group Table 3-12 Pharmacokinetic parameters of ampicillin in high-dose test group

t _1/2 AUC _0-t AUC _0-∞ t _1/2 AUC _0-t AUC _0-∞

High Dose-R High Dose-T

(h) (μg·h·mL ^-1 ) (μg·h·mL ^-1 ) (h) (μg·h·mL ^-1 ) (μg·h·mL ^-1 )

1 1.61 250.8 251.1 1 1.71 269.9 270.7

2 1.59 191.8 192.1 2 1.98 237.4 238.0

3 1.34 199.8 200.1 3 1.46 238.7 239.0

4 1.27 200.3 200.5 4 1.53 196.5 196.8

5 1.52 244 244.3 5 1.88 336.9 337.9

6 1.47 331.5 331.9 6 1.73 425.3 426.2

Mean 1.49 233.7 234 Mean 1.75 275.5 276.1

SD 0.13 48.71 48.78 SD 0.21 79.55 79.77

      Table 3-1 Plasma concentration of ampicillin in the low-dose control group at different times (g·mL-1)

                                             

Time(h)

1 2 3 3 4 5 5 6 Mean SD

0.17 115.2 161.0 122.0 121.0 108.0 125.0 125.4 18.47

0.50 78.70 92.9 67.9 69.6 61.3 68.30 73.1 11.18

1.00 35.60 53.3 40.9 36.7 31.9 35.30 39.0 7.60

1.50 26.40 37.5 22.5 21.9 16.8 29.60 25.8 7.19

2.00 18.10 24.3 12.6 11.4 9.23 15.00 15.1 5.44

3.00 5.65 12.8 5.24 5.17 3.82 4.30 6.16 3.32

4.00 2.25 5.65 1.77 1.88 1.31 1.56 2.40 1.62

6.00 0.54 0.99 0.18 0.10 0.12 0.33 0.38 0.34

8.00 n.d. 0.18 n.d. n.d. n.d. n.d. 0.03 0.07

n.d.: below the lower limit of quantitation

Table 3-2 Plasma concentration of ampicillin in the low-dose test group at different times (μg·mL ^-1 )

                                             

Time(h)

1 2 3 3 4 5 6 6 Mean SD

0.17 123.0 152.0 124.0 133.0 143.0 142.0 136.2 11.51

0.50 76.6 85 68.9 78.4 74.7 63.7 74.6 7.46

1.00 44.3 50.8 45.8 49.3 55.9 36.9 47.2 6.47

1.50 32.9 33.3 29.6 28 33.4 25.1 30.4 3.41

2.00 18.2 25.8 20.0 24.8 23.8 14.4 21.2 4.42

3.00 7.86 16.0 10.7 11.8 13.7 10.2 11.71 2.85

4.00 7.06 8.00 5.75 5.50 5.90 4.66 6.15 1.19

6.00 1.44 2.45 1.18 1.24 1.36 1.59 1.54 0.47

8.00 0.37 0.55 0.23 0.30 0.34 0.58 0.40 0.14

10.0 0.10 0.12 n.d. n.d. n.d. 0.12 0.06 0.06

n.d.: below the lower limit of quantitation

The blood concentration of ampicillin in the dose control group at different times in Table 9 (μg·mL ^-1 )

                                     

Time(h)

1 2 3 3 4 5 6 6 Mean SD

0.17 134.6 127.6 89.00 149.8 188.6 144.8 139.1 32.43

0.50 80.20 73.20 60.80 73.20 98.00 98.40 80.6 14.98

1.00 42.40 42.20 34.60 61.80 54.00 53.80 48.1 10.05

1.50 24.60 28.80 22.40 33.60 29.40 28.60 27.9 3.93

2.00 12.78 18.10 11.90 18.04 15.72 12.76 14.9 2.79

3.00 3.40 6.66 2.40 5.50 4.30 2.80 4.20 1.65

4.00 2.33 4.31 2.25 2.83 2.95 2.35 2.80 0.78

6.00 0.46 0.97 0.42 0.64 0.43 0.47 0.60 0.21

8.00 0.11 0.21 0.10 n.d. n.d. n.d. 0.07 0.09

n.d.: below the lower limit of quantitation

The plasma concentration of ampicillin at different times in the dosage test group in Table 10 (μg·mL ^-1 )

Time(h) Animal ID

1 2 3 3 4 5 5 6 Mean SD

0.17 151.2 106.4 98.20 182.6 181.4 161.4 146.9 36.62

0.50 81.00 57.00 54.60 115.8 100.4 95.60 84.07 24.56

1.00 45.80 39.40 37.00 75.00 63.00 66.80 54.50 15.84

1.50 29.20 28.00 28.20 46.40 40.80 42.00 35.77 8.22

2.00 17.20 18.50 18.06 24.80 27.20 26.40 22.03 4.58

3.00 5.92 7.0 6.1 9.18 9.92 11.4 8.25 2.24

4.00 3.43 6.53 4.68 5.16 8.16 9.36 6.22 2.24

6.00 0.76 1.68 1.21 1.35 2.00 2.52 1.59 0.62

8.00 0.19 0.59 0.31 0.41 0.72 0.81 0.51 0.24

10.0 n.d. 0.13 n.d. 0.22 0.21 0.22 0.13 0.11

n.d.: below the lower limit of quantitation

Table 11 Plasma concentration of ampicillin in the high-dose control group at different times (μg·mL ^-1 )

Animal ID

Time(h)

1 2 3 3 4 5 6 6 Mean SD

0.17 197.6 179.6 193.6 188.4 225.2 287.6 212.0 40.11

0.50 126.4 94.4 130.8 117.2 138.8 174 130.3 26.30

1.00 79.20 63.20 49.20 67.60 78.40 98.40 72.67 16.76

1.50 54.40 38.00 39.68 41.20 47.20 72.00 48.75 12.88

2.00 33.92 21.04 24.68 20.72 26.96 46.00 28.89 9.67

3.00 19.16 11.40 8.70 8.07 12.40 19.50 13.21 5.01

4.00 7.64 3.99 2.65 3.00 4.73 7.89 4.98 2.28

6.00 1.76 0.86 0.45 0.52 0.93 1.43 0.99 0.51

8.00 0.44 0.25 0.19 0.20 0.34 0.55 0.33 0.15

10.0 0.21 0.16 0.11 0.11 0.17 0.26 0.17 0.06

12.0 0.13 0.12 n.d. n.d. 0.12 0.17 0.09 0.07

n.d.: below the lower limit of quantitation

Table 12 Plasma concentration of ampicillin in the high-dose test group at different times (μg·mL ^-1 )

Animal ID

Time(h)

1 2 3 3 4 5 6 6 Mean SD

0.17 210.4 194.8 176.4 169.2 266 337.6 225.7 64.74

0.50 120.4 127.2 142.0 78.80 164.4 200.8 138.9 41.43

1.00 97.20 66.40 64.40 60.80 107.2 134.0 88.33 29.43

1.50 54.40 43.20 48.40 33.88 72.00 90.00 56.98 20.59

2.00 35.04 34.80 35.40 27.16 33.96 52.80 36.53 8.55

3.00 11.72 14.80 15.32 15.30 26.12 30.32 18.93 7.44

4.00 11.88 8.28 8.84 7.06 14.16 17.00 11.20 3.85

6.00 3.18 1.93 2.64 2.10 3.36 3.99 2.87 0.79

8.00 2.07 1.03 0.69 0.78 1.24 1.49 1.22 0.51

10.0 0.50 0.48 0.26 0.36 0.59 0.66 0.47 0.15

12.0 0.34 0.24 0.15 0.13 0.37 0.36 0.27 0.11

n.d.: below the lower limit of quantitation

4. Discussion

The study found that the blood elimination half-life t _1/2β of ampicillin at low, medium and high dose levels in the test group was longer than that of the control group, prolonging 0.28, 0.23 and 0.26h respectively; the AUC _0-t and AUC of ampicillin in the test group _0- also improved compared to the control group by 23.4%, 20.3% and 17.9%, respectively. The above data were statistically processed, and the differences were significant (P<0.05). The AUC _0-t and AUC _0-∞ of the three dose groups were linearly correlated with the dose (r>0.9708, P<0.05). This shows that probenecid sodium (potassium) can obviously prolong the half-life of ampicillin sodium and improve blood drug concentration, so that the total amount of medicine in the body in the unit time of ampicillin has also been increased accordingly, so the drug effect will be obviously improved. It is particularly important that probenecid can significantly prolong the time for ampicillin sodium to exceed the MIC of the corresponding bacteria, which is not only beneficial to kill bacteria, but also more conducive to reducing the occurrence and development of drug resistance, because when the concentration of the drug is lower than the MIC It is very easy to lead to the generation of drug-resistant strains.

Effect of Probenecid Sodium (Potassium) on Elimination Half-life (t _1/2β ) of Penicillin G Sodium, Cefazolin Sodium, Cefuroxime Sodium, Cefotaxime Sodium and Cefoxitin Sodium

1. Purpose: This test aims to observe the blood elimination half-life (t _1/2β ) of probenecid sodium (potassium) on penicillin G sodium salt, cefazolin sodium, cefuroxime sodium, cefotaxime sodium and cefoxitin sodium. The impact laid the foundation for the research and development of a series of compound prescriptions.

2. Test materials

2.1 Drugs and reagents

Probenecid Sodium (Potassium)/ Penicillin G Sodium Salt, Probenecid Sodium (Potassium)/ Cefazolin Sodium, Probenecid Sodium (Potassium)/ Cefuroxime Sodium, Probenecid Sodium (Potassium)/ Cefotaxime Sodium and probenecid sodium (potassium)/cefoxitin sodium compound prescription powder (pencillin bottle) and corresponding antibiotic reference substance; enalapril maleate (internal standard, purity provided by Beijing Institute for the Control of Pharmaceutical and Biological Products: 99.8%) ; Canine blank plasma (provided by the Department of Experimental Animals, School of Basic Medicine, Jilin University).

Methanol was chromatographically pure and was purchased from Tianjin Concord Technology Co., Ltd. Other reagents were analytically pure and were products of Tianjin Fuchen Chemical Reagent Factory and Beijing Chemical Factory.

2.2 Test animals

Adult Beagle dogs with a weight range of 10.50±1.3 kg were purchased from the Experimental Animal Center of Sichuan Academy of Medical Sciences.

2.3 Experimental program and administration method

Beagle dogs were randomly divided into groups (control group R and test group T), and a two-period crossover test was carried out, and the interval between two administration cycles was 7 days. The dosage and method of administration are as follows:

Control group: Each dog in each control group was individually given the corresponding antibiotic control substance, dissolved in normal saline, and then intravenously injected. The specific doses are: penicillin G: 80.65 mg/kg, cefazolin: 53.76 mg/kg, cefuroxime: 40.32 mg/kg, cefotaxime: 40.32 mg/kg, cefoxitin: 53.76 mg/kg.

Experimental group: each dog in each experimental group was given the corresponding antibiotic and probenecid sodium (potassium) mixture, dissolved in normal saline, and then injected intravenously. The specific dose was: each dog was given the same antibiotics as the above-mentioned control group, and at the same time, 26.88 mg/kg of probenecid was injected intravenously.

2.4 Sample collection and treatment

About 1ml of blood was collected from the small saphenous vein of the canine hindlimb. Immediately after the blood sample was taken out, it was transferred into a heparin test tube, centrifuged (3500rpm) for 10min, the plasma was separated, and stored frozen in a -20°C refrigerator until assayed.

2.5 Plasma sample analysis method

Canine plasma samples were determined by LC/MS/MS method.

2.6 Data processing

Compare and examine whether there is any statistical difference in the blood elimination half-life (t _1/2β ) between the experimental group and the control group.

3. Results

The impact of table 1 probenecid sodium (potassium) on the blood elimination half-life of different β-lactam antibiotics Blood elimination half-life (hours, probenecid sodium group) Blood elimination half-life (hours, probenecid potassium group) control group test group control group test group Penicillin G 0.48 1.26 0.51 1.22 Cefazolin 1.36 2.23 1.32 2.31 Cefuroxime 0.72 1.21 0.68 1.28 Cefotaxime 1.48 2.13 1.42 2.26 Cefoxitin 0.68 1.36 0.71 1.31

Note: There is no statistical difference between the probenecid sodium group and the probenecid potassium group, but the difference between the control group and the test group is significant P<0.05.

4. Discussion

The study found that probenecid sodium (potassium) significantly prolonged the blood elimination half-life (t1/2β) of antibiotics in the test group. Since the above-mentioned β-lactam antibiotics are all time-dependent antibiotics, the most critical factor for the antibacterial effect of time-dependent antibiotics is not the level of the peak blood concentration, but the key is whether the blood concentration exceeds the corresponding pathogenic bacteria. MIC. Probenecid sodium (potassium) prolongs the half-life of the above-mentioned antibiotics, which means that probenecid sodium (potassium) can significantly prolong the time when the blood concentration of the above-mentioned antibiotics exceeds the MIC of the corresponding pathogenic bacteria. This shows that probenecid sodium (potassium) can also form a compound preparation with the above-mentioned β-lactam antibiotics. In addition, there was no significant difference between the sodium and potassium salts of probenecid in prolonging the effects of the above antibiotics.

The positive effects of the present invention are: probenecid sodium (potassium) and β-lactam antibiotics are intravenously injected or intravenously dripped at the same time, which increases the total amount of medicine in the body of β-lactam antibiotics per unit time, and can obviously improve efficacy. At the same time, it can also reduce the dosage of antibiotics and prolong the interval of administration, which not only saves resources but also facilitates patients. It is safe, has no toxic and side effects, and has good medicinal prospects; the preparation process is simple and the use is more convenient. Reduce the occurrence and development of bacterial resistance, save antibiotic resources, facilitate patients, and conform to the principles of medical economics

Detailed ways:

The present invention will be further described below in conjunction with embodiment:

Example 1: Dissolve 22 kilograms of sodium hydroxide in 200 liters of purified water, add 142 kilograms of probenecid after all the dissolution, heat to 60° C., adjust the pH to above 7.5, and keep stirring until the pH value is basically constant. Under the condition of 60°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions to obtain the raw material of probenecid sodium that meets the standards for human drug injections. White or off-white crystalline powder.

Probenecid sodium and penicillin G sodium are mixed under aseptic conditions at a weight ratio of 1:3, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 2: Dissolve 28 kg of sodium hydroxide in 280 liters of purified water, add 143 kg of probenecid after all the dissolution, heat to 65°C, adjust the pH to above 8, and keep stirring until the pH value is basically constant. Under the condition of 65°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions to obtain the raw material of probenecid sodium that meets the standards for human drug injections. White or off-white crystalline powder.

Probenecid sodium and ampicillin sodium are mixed under aseptic conditions at a weight ratio of 1:2.8, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 3: Dissolve 40 kg of sodium hydroxide in 300 liters of purified water, add 150 kg of probenecid after all dissolved, heat to 70°C, adjust pH to 10, and keep stirring until the pH value is basically constant. Under the condition of 70°C, it is sterilized by filtration through a 0.45 μm filter membrane, then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions to obtain the raw material of probenecid sodium that meets the standards for human drug injections. White or off-white crystalline powder.

Probenecid sodium and cefazolin sodium are mixed under aseptic conditions at a weight ratio of 1:2, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 4: first dissolve 102 g of sodium hydroxide in 130 g of purified water (or water for injection) and completely dissolve, add 663 g of probenecid, heat to 68 ° C, adjust the pH to 9, and keep stirring until the pH value is basically constant; Sterilize by filtering through a 0.45μm filter membrane at ℃, then finely filter through a 0.22μm filter membrane to remove pyrogens, and dry under sterile conditions to obtain the raw material of probenecid sodium that meets the standards for human drug injections, white or Off-white crystalline powder;

Probenecid sodium and cefuroxime sodium are mixed under aseptic conditions at a weight ratio of 1:1.5, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 5: First, dissolve 0.17% potassium hydroxide in 11.5% purified water (or water for injection), add 0.88% probenecid, heat to 60°C, adjust the pH to 7.5, and keep stirring until the pH value is basically constant; Under the condition of 0.45 μm filter membrane to filter and sterilize, then pass through a filter membrane of 0.22 μm for fine filtration to remove pyrogens, and dry under sterile conditions to obtain the probenecid potassium raw material that meets the standards for human medicine injections. It is white or Off-white crystalline powder; mix probenecid potassium and cefotaxime sodium at a weight ratio of 1:1.5 under aseptic conditions, subpackage under aseptic conditions, and finally make a powder injection.

Example 6: First, dissolve 173.44 potassium hydroxide in 1308 purified water (or water for injection) and completely dissolve it, add 885.75 probenecid, heat to 70°C, adjust the pH to 10, and keep stirring until the pH value is basically constant; at 70 Under the condition of 0.45 μm filter membrane to filter and sterilize, then pass through a filter membrane of 0.22 μm for fine filtration to remove pyrogens, and dry under sterile conditions to obtain the probenecid potassium raw material that meets the standards for human medicine injections. It is white or Off-white crystalline powder; mix probenecid potassium and cefoxitin sodium at a weight ratio of 1:2 under aseptic conditions, subpackage under aseptic conditions, and finally make a powder injection.

Example 7: Dissolve 22 kg of potassium hydroxide in 200 liters of purified water, add 142 kg of probenecid after all dissolved, heat to 60° C., adjust pH to above 7.5, and keep stirring until the pH value is basically constant. Under the condition of 60°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions to obtain the probenecid potassium raw material that meets the standards for human drug injections, as White or off-white crystalline powder.

Probenecid potassium and penicillin G sodium are mixed under aseptic conditions at a weight ratio of 1:3, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 8: Dissolve 28 kg of potassium hydroxide in 280 liters of purified water, add 143 kg of probenecid after all the dissolution, heat to 65°C, adjust the pH to above 8, and keep stirring until the pH value is basically constant. Under the condition of 65°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions to obtain the probenecid potassium raw material that meets the standards for human drug injections. White or off-white crystalline powder.

Probenecid potassium and ampicillin sodium are mixed under aseptic conditions at a weight ratio of 1:2.8, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 9: Dissolve 40 kg of potassium hydroxide in 300 liters of purified water, add 150 kg of probenecid after all dissolved, heat to 70° C., adjust pH to 10, and keep stirring until the pH value is basically constant. Under the condition of 70°C, it is sterilized by filtration through a 0.45 μm filter membrane, and then finely filtered through a 0.22 μm filter membrane to remove pyrogens, and dried under sterile conditions to obtain the probenecid potassium raw material that meets the standards for human drug injections, as follows: White or off-white crystalline powder.

Probenecid potassium and cefazolin sodium are mixed under aseptic conditions at a weight ratio of 1:2, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 10: First, dissolve 102% potassium hydroxide in 1308% purified water (or water for injection), add 663% probenecid, heat to 68°C, adjust the pH to 9, and keep stirring until the pH value is basically constant; Under the condition of 0.45 μm filter membrane to filter and sterilize, then pass through a filter membrane of 0.22 μm for fine filtration to remove pyrogens, and dry under sterile conditions to obtain the probenecid potassium raw material that meets the standards for human medicine injections. It is white or Off-white crystalline powder;

Probenecid potassium and cefuroxime sodium are mixed under aseptic conditions at a weight ratio of 1:1.5, subpackaged under aseptic conditions, and finally made into a powder injection.

Example 11: First, dissolve 0.17% sodium hydroxide in 11.5% purified water (or water for injection), add 0.88% probenecid, heat to 60°C, adjust the pH to 7.5, and keep stirring until the pH value is basically constant; Sterilize by filtering through a 0.45μm filter membrane at ℃, then finely filter through a 0.22μm filter membrane to remove pyrogens, and dry under sterile conditions to obtain the raw material of probenecid sodium that meets the standards for human drug injections, white or Off-white crystalline powder; mix probenecid sodium and cefotaxime sodium at a weight ratio of 1:1.5 under aseptic conditions, subpackage under aseptic conditions, and finally make a powder injection.

Example 12: First, dissolve 173.44 sodium hydroxide in 1308 purified water (or water for injection) and completely dissolve it, add 885.75 probenecid, heat to 70°C, adjust the pH to 10, and keep stirring until the pH value is basically constant; at 70 Sterilize by filtering through a 0.45μm filter membrane at ℃, then finely filter through a 0.22μm filter membrane to remove pyrogens, and dry under sterile conditions to obtain the raw material of probenecid sodium that meets the standards for human drug injections, white or Off-white crystalline powder; mix probenecid sodium and cefoxitin sodium at a weight ratio of 1:2 under aseptic conditions, and subpackage under aseptic conditions, and finally make a powder injection.

Claims (7)

1. The structural formula of probenecid sodium is as follows:

the molecular formula is: c₁₃H₁₈NO₄The molecular weight of SNa is: 307.34

The structural formula of probenecid potassium:

the molecular formula is: c₁₃H₁₈NO₄The molecular weight of K is: 323.45

2. The process for preparing probenecid according to claim 1, wherein the process comprises the following steps: firstly, dissolving 0.9-102 sodium hydroxide in 11.5-1308 purified water, adding 6.5-663 probenecid after all the sodium hydroxide is dissolved, heating to 60 +/-10 ℃ to ensure that the probenecid completely reacts; adjusting the pH value to 7.5-10, filtering and sterilizing through a 0.45 mu m filter membrane at the temperature of 60 +/-10 ℃, then performing fine filtration through a 0.22 mu m filter membrane to remove pyrogen, and drying under the aseptic condition to obtain the probenecid sodium raw material which is white or white-like crystalline powder and meets the human injection standard. The probenecid content of the product is not lower than 88.50% calculated on a dry product.

3. The process for preparing probenecid potassium according to claim 1, characterized in that: firstly, dissolving 0.17-173.44 potassium hydroxide in 11.5-1308 purified water, adding 0.88-885.75 probenecid after all the potassium hydroxide is dissolved, heating to 60 +/-10 ℃ to ensure that the probenecid completely reacts; adjusting the pH value to 7.5-10, filtering and sterilizing through a 0.45 mu m filter membrane at the temperature of 60 +/-10 ℃, then carrying out fine filtration through a 0.22 mu m filter membrane to remove pyrogen, and drying under the aseptic condition to obtain the probenecid potassium raw material which is white or white-like crystalline powder and meets the human injection standard. The probenecid content of the product is not lower than 86.25 percent calculated on a dry product.

4. The compound injection of claim 1, wherein the injection comprises probenecid sodium, probenecid potassium and β -lactam antibiotics, and is prepared by mixing probenecid sodium, probenecid potassium and β -lactam antibiotics at a ratio of 1: 1-3, packaging under aseptic condition, and making into powder for injection and lyophilized powder for injection.

5. The compound injection of probenecid sodium, probenecid potassium and β -lactam antibiotics of claim 4, wherein the powder for injection and the lyophilized powder for injection are simultaneously injected or instilled intravenously with β -lactam antibiotics.

6. The compound injection of claim 4, wherein the probenecid sodium and probenecid potassium and the β -lactam antibiotics comprise penicillin antibiotics and cephalosporin antibiotics, the penicillin antibiotics comprise penicillin G and ampicillin, and the cephalosporin antibiotics comprise cefazolin, cefuroxime, cefotaxime and cefoxitin.

7. Compound injection composed of probenecid sodium (potassium) and β -lactam antibiotics and its useUse for prolonging the blood elimination half-life (t) of β -lactam antibiotics_1/2β) Increasing the area under the curve (AUC) and significantly prolonging the time during which the blood concentration of β -lactam antibiotic exceeds the MIC of the corresponding bacterium.