CN116023284B - Addition salts of propofol derivatives with pharmaceutically acceptable acids, compositions and uses thereof - Google Patents

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a low-toxicity propofol derivative salt, a composition and application thereof. The in vitro screening test results show that the hemolysis of the addition salt disclosed by the invention is obviously lower than that of 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyric acid ester hydrochloride disclosed in CN106132922A, and the respiratory depression effect is weakened, so that the safety is obviously improved.

Description

Addition salts of propofol derivatives with pharmaceutically acceptable acids, compositions and uses thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a low-toxicity propofol derivative salt, a composition and application thereof.

Background

According to the "national drug administration" in regard to the registration classification of issued chemicals and the announcement of requirements of the claim materials (No. 44 in 2020) (No. 20 in 6 in 2020), the 2.1 new drug means "a drug which contains an optical isomer of a known active ingredient prepared by resolution or synthesis, or forms an ester of the known active ingredient, or forms a salt of the known active ingredient (including a salt containing a hydrogen bond or a coordinate bond), or changes an acid group, a base or a metal element of the known salt active ingredient, or forms other noncovalent bond derivatives (such as a complex, a chelate or an inclusion compound), and has remarkable clinical advantages. Wei Minji et al, in the "drug registration management methods" the problem thinking about human pharmacokinetics of 3 and 4 classes of drugs "were published (in China, J.New drug 2014,23 (19): 2219-2223.)" if the therapeutic window of the drug was narrow, acid groups or bases (or metal recommended elements) might affect the pharmacologically active moiety, it was necessary to consider the point of view of the effectiveness and safety in the 4.1 test design. Among these, the concept of "pharmacological activity" includes toxicological effects, such as Zheng Xinting, in the "observations of effects of toluene sulfonic acid Rayleigh Malun on sedative analgesia in the elderly colonoscopy" (in northern medicine, 2021,18 (02): 35-36), indicates that toluene sulfonic acid Rayleigh Malun is less toxic than benzene sulfonic acid Ramazole.

Zhang Daping et al, in the "salt form selection of drugs in New drug development" (journal of the Chinese medical industry, 2011,42 (08): 631-635.) "indicate" from the pH-solubility relationship of drugs, which counter ions can form salts with drugs, the ease of conversion to the corresponding acids or bases in the physiological pH range after salt formation, whether the solubility and dissolution rate are affected by the same ions, indicate that salt-type compounds are not necessarily immediately converted to the corresponding free acids/bases in aqueous solution ".

CN106132922a discloses a series of water-soluble prodrugs of propofol and pharmaceutically acceptable salts thereof, and pharmacological test results show that the hydrochloride or sodium salt of the water-soluble prodrug is specific for rat and mouse LD ₅₀ In the range of 20 to 80mg/kg, the present inventors found that the hydrochloride of 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyric acid ester had a certain hemolysis, and the safety profile thereof was still further improved.

Disclosure of Invention

The object of the present invention is to provide an addition salt of a propofol derivative with a pharmaceutically acceptable acid, which salt has significantly less haemolysis than the salt disclosed in CN106132922a, as well as a composition and use thereof.

In order to achieve the above object, the present invention provides, in one aspect, an addition salt of 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyric acid ester with a pharmaceutically acceptable acid.

In one aspect, the pharmaceutically acceptable acid of the present invention is one selected from succinic acid, tartaric acid, sulfuric acid, p-toluenesulfonic acid, nitric acid, phosphoric acid, methanesulfonic acid, citric acid, malic acid, fumaric acid, maleic acid and benzoic acid.

In another aspect, the salt of the present invention preferably has a molar ratio of 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyric acid ester to pharmaceutically acceptable acid of 1:1.

In another aspect the invention provides a composition comprising an addition salt as hereinbefore described.

In one aspect, preferably, the compositions of the present invention are formulated as liquid formulations; further preferably, the liquid formulation is sterile.

In another aspect, preferably, the composition of the present invention may be formulated as a powder for injection; further preferably, the injectable powder of the present invention is lyophilized.

In a further aspect the invention provides the use of an addition salt or composition as hereinbefore described in the manufacture of an anaesthetic.

The in vitro screening assay showed that the addition salts of the present invention showed significantly lower hemolysis in canine blood than the 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyrate hydrochloride disclosed in CN106132922 a.

Drawings

FIGS. 1-3 Compounds A phosphate, sulfate and nitrate ¹ H-NMR chart

FIGS. 2-1 to 2-5 are the results of the hemolysis test.

Detailed Description

Example 1 preparation and Structure characterization of addition salts

Preparation method

0.1mol of 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyric acid ester (compound A) hydrochloride prepared by the method disclosed in CN106132922A was taken and suspended in 100mL of isopropyl acetate, and then 0.1mol of sodium bicarbonate (saturated aqueous solution) was added thereto, and the mixture was stirred at room temperature until the mixture was dissolved. Standing for separating, separating organic phase, concentrating under reduced pressure to about 1/5 of original volume, adding 0.1mol of acid respectively, stirring overnight, and making TLC complete reaction, wherein the reaction solution is neutral. Filtering, crystallizing with isopropyl acetate, filtering, and drying to constant weight.

The number of addition salts prepared using different acids and the yield and purity data are shown in table 1.

TABLE 1

Structural characterization

Phosphates, nitrates and sulfates ¹ The H-NMR chart and data and the results of elemental analysis are shown in Table 2.

TABLE 2

As shown in the table 2 below,even phosphoric, sulfuric and nitric acids, which are relatively highly acidicNeither compound a is degraded, but an acid addition salt of a is formed.

Elemental analysis results showed that compound a was mixed with either phosphoric acid, sulfuric acid or nitric acid at 1:1 in a molar ratio of salt formation.

The elemental analysis results of other acid addition salts of Compound A are shown in Table 3

TABLE 3 Table 3

EXAMPLE 2 hemolysis test

Test example 1: hemolysis test 1 of addition salts

1. Sample: 337API-1, 337API-2, 337API-3, 337API-4, 337API-5, 337API-6 shown in Table 1, the hydrochloride of Compound A was taken as 337API-7.

2. Materials and instruments: 0.9% sodium chloride injection, 2% erythrocyte suspension (canine blood), a medical centrifuge and a constant-temperature water bath kettle.

3. The test method comprises the following steps: the samples are prepared into concentrations of 77.6mg/mL, 38.8mg/mL, 19.4mg/mL, 9.7mg/mL, 4.85mg/mL and 2.425mg/mL by using 0.9% sodium chloride injection, 5 clean glass test tubes are used for each concentration, 1 and 2 tubes are used as test sample tubes, 3 tubes are used as negative control tubes, 4 tubes are used as positive control tubes, 5 tubes are used as test sample control tubes, 2% red cell suspension, 0.9 sodium chloride injection and pure water are sequentially added as shown in the table, and after uniform mixing, the mixture is immediately incubated in a constant-temperature water bath kettle at 37+/-0.5 ℃. After 3 hours, hemolysis and coagulation reactions were observed.

4. Test results

The hemolysis test results are shown in the accompanying figures 2-1 to 2-5 and Table 2.

TABLE 2

Note that: "+" indicates hemolysis, "-" indicates non-hemolysis, "+" indicates slight hemolysis, and "++" indicates hemolysis and coagulation

As can be seen from FIGS. 2-1 to 2-5 and Table 2, the hemolysis of the phosphate of Compound A is lower than that of the other addition salts at a sample concentration of 19.4mg/mL or less.

Test example 2 hemolysis test 2 of acid addition salt

Test examples 1 were the same as test examples 1 except that 337API-8 to 337API-13 and 337API-7 shown in Table 1 were used as the test samples.

The results are shown in Table 3.

TABLE 3 Table 3

Test example 3 anesthetic action of acid addition salts

1. Sample: the acid addition salts shown in Table 1 were prepared in a concentration of 12.5mg/mL using water for injection as a solvent.

2. Test animals: beagle dogs, 52 animals, each half of which is female and male, weight 7-12 kg, are equally divided into 13 groups, each group of which is female and male.

3. The method comprises the following steps: each sample was given to Beagle dogs by single intravenous injection at a rate of 6.0mL/min at an anesthesia induction dose of 15mg/kg (12.50.+ -. 1.88 mg/kg), and the incubation time for induction of anesthesia, the maintenance of anesthesia time and the wake-up standing time were recorded.

4. Results: as shown in table 4 (time units s).

The results in table 4 show that: the various addition salts provided by the invention maintain good anesthetic effect.

Example 3 respiratory depression assay

1. Sample: the acid addition salts and propofol shown in Table 1 were prepared in a solution having a concentration of 12.5mg/mL using water for injection as the vehicle.

2. Test animals: 60 healthy Beagle dogs are half-divided into a male and a female, the ages of the male and the female are 2-4 years, the weight of the healthy Beagle dogs is 10-15 kg, and the healthy Beagle dogs are equally divided into 15 groups, and each group is half-divided into the male and the female.

3. The method comprises the following steps: the test animals were fasted for 8h, water was forbidden for 4h, weighed, and venous access was established prior to anesthesia. The blank group did not perform any operation; the other groups were given to Beagle dogs by intravenous injection, and were first subjected to anesthesia induction by 15mg/kg (12.50.+ -. 1.88 mg/kg) intravenous injection, the anesthesia induction rate was 6.0mL/min, the blank group was not subjected to any operation, and the other groups were then subjected to anesthesia maintenance by 20 mg/kg.h intravenous injection of the test sample. And monitoring physiological indexes such as respiration, blood oxygen saturation and the like of the dogs by adopting an iPM12Vet electrocardiograph monitor. Each group was monitored 1 time and recorded at 15min, 30min, and 60min after 1 time of physiological index as a base value before anesthesia. Finally, the physiological indexes such as respiration, blood oxygen saturation and the like of the tested animal within 60 minutes of injection are obtained.

4. The results are shown in tables 5 and 6

TABLE 5 monitoring results of respiratory rate of dogs in each group (times/min) during anesthesia practice

Table 6 monitoring results of blood oxygen saturation of groups of dogs during anesthesia practice (unit:%)

Table 5 shows the results of monitoring the respiratory rate of each group of dogs during the course of anesthesia administration. During anesthesia, the respiratory rate of the animals in the groups other than 337API-6 and 337API-9 differed little. Wherein the respiratory rate of the tested animals injected with the 337API-6 group is far higher than that of the other groups, and the respiratory rate of the tested animals injected with the 337API-9 group is also significantly higher than that of the other groups, which indicates that the phosphate and the citrate of the invention have the effect of significantly reducing respiratory depression.

Table 6 shows the results of monitoring blood oxygen saturation of dogs during anesthesia practice. From the blood oxygen saturation data of each group of test animals, the blood oxygen saturation of the test animals of the 337API-6 group is far higher than that of the test animals of other groups, and the blood oxygen saturation of the test animals of the 337API-9 group is also significantly higher than that of the other groups, which is related to the respiratory rate of the test animals, so that the product with the product number of 337API-6 is strongly proved to be capable of significantly reducing respiratory depression.

In conclusion, the respiratory depression experiments prove that the phosphate and the citrate have unexpected reduced respiratory depression effect and are safer. The propofol has obvious inhibition effect on respiration, can inhibit the ventilation reaction of carbon dioxide, is shown by the reduction of tidal volume, can increase the respiratory frequency in a waking state, has no obvious influence on bronchial smooth muscle due to the frequent apnea caused by intravenous injection. The discovery has remarkable significance, and the speculated mechanism is probably that the PI3K/Akt signal pathway is an important pathway for membrane receptor signal transmission into cells, and extracellular signal stimulation such as cytokines, growth factors, hormones and the like can promote PI3K activation, and the PI3K activation can phosphorylate Ser473 site of Akt to activate Akt. The test substance prepared by the invention can activate the expression of P13 protein, thereby activating the expression of Akt protein. The Akt protein can inhibit apoptosis of lung cells, thereby affecting respiratory function of the lung and achieving the effect of reducing respiratory inhibition.

EXAMPLE 4 preparation of a sterile solution containing Compound A phosphate

1000g of compound A phosphate is taken, 10 times of water for injection is added, heating reflux is carried out for 6 hours, distillation is carried out, 6000mL of primary distillate is collected, and then double distillation is carried out until 1000mL. After content measurement, 8.5g of sodium chloride and 80 mL of polysorbate are added to dissolve all, and the mixture is filtered, encapsulated and sterilized at 100 ℃ for 30 minutes to obtain the finished product.

EXAMPLE 5 preparation of sterile lyophilized powder for injection containing Compound A phosphate

Taking 10mg of compound A phosphate, 10mg of mannitol, 5mg of hydrolyzed gelatin and 1g of calcium gluconate, dissolving in a proper amount of water for injection, carrying out sterile filtration, subpackaging in ampoule bottles with 0.5mL of each, freeze-drying, sealing, and checking air leakage.

Claims (7)

1. Addition salts of 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyrate with pharmaceutically acceptable acids;

   the pharmaceutically acceptable acid is one selected from phosphoric acid and citric acid;

   the molar ratio of the 2, 6-diisopropylphenyl (R) -4- (dimethylamino) -2-fluorobutyric acid ester to the pharmaceutically acceptable acid in the salt is 1:1.
2. 2. A composition comprising the addition salt of claim 1.
3. 3. The composition according to claim 2, wherein the composition is formulated as a liquid formulation.
4. 4. A composition according to claim 3, characterized in that the liquid formulation is sterile.
5. 5. The composition according to claim 2, wherein said composition is formulated as a powder for injection.
6. 6. The composition of claim 5, wherein the injectable powder is lyophilized.
7. 7. Use of an addition salt according to claim 1 or a composition according to claim 2 for the preparation of an anesthetic.