CN111346093A

CN111346093A - Application of combination of lamivudine and domperidone in preparation of sedative drugs

Info

Publication number: CN111346093A
Application number: CN202010325570.3A
Authority: CN
Inventors: 颜晓萍; 戴云翔; 李嫣妮; 郑文忠
Original assignee: Zhangzhou Health Vocational College
Current assignee: Zhangzhou Health Vocational College
Priority date: 2020-04-23
Filing date: 2020-04-23
Publication date: 2020-06-30
Anticipated expiration: 2040-04-23
Also published as: CN111346093B

Abstract

The invention provides application of combination of lamivudine and domperidone in preparing a sedative drug. The research of the invention finds that when the lamivudine and the domperidone are combined in a specific proportion, the lamivudine and the domperidone have exact sedative effect; moreover, its sedative effect is relatively mild and does not cause loss of communication ability or difficulty in arousal. Further pharmacological property research shows that the medicine can promote GABA and GABA_AReceptor binding, increasing the frequency of chloride channel opening; at the same timeRegulating the content of glutamic acid in cerebral cortex and hypothalamus of experimental animals, thereby relieving excitable neurotoxicity in brain; moreover, the inhibitor can be selectively combined with 5-hydroxytryptamine 1A receptor, inhibit the synapse from absorbing 5-hydroxytryptamine, and thus up-regulate the content of 5-hydroxytryptamine. Based on the above new properties discovered, the invention determines the new application of the combination of lamivudine and domperidone in preparing the sedative, not only expands the selectable range of the sedative, but also improves the sedative effect to a certain extent.

Description

Application of combination of lamivudine and domperidone in preparation of sedative drugs

Technical Field

The invention relates to the technical field of medicinal chemistry, and further relates to a novel medical application of a substance, in particular to an application of lamivudine and domperidone in preparation of a sedative drug.

Background

Sedative is a general term for a large class of medicines for promoting human bodies to calm and sleep, is mainly used for symptomatic treatment of symptoms such as mental disorder anxiety, dysphoria and the like, and the ideal sedative medicine needs to have the following characteristics: 1. has little effect on circulation and respiration: 2. the effect is quick and obvious; 3. the drug metabolism is fast, and basically no obvious accumulation function exists: 4. the influence on the liver and kidney functions is small; 5. the anxiolytic and the compliance forgetting effect are excellent, the easy awakening during the use, the rapid recovery of consciousness state after stopping taking medicine, and the like, at present, the common sedative mainly comprises benzene: azepines, phenothiazines, butyrophenones, and the like.

Benzodiazepines are ideal sedative hypnotic drugs, and are commonly used as midazolam, diazepam and lorazepam. Diazepam has good anxiolytic effect under small dosage, does not affect consciousness, can cause lethargy and loss of consciousness when injected with larger dosage, has no obvious influence on respiration, can generate apnea when used together with other central inhibitory drugs, and can lower blood pressure, dilate coronary artery and increase coronary blood flow when used with large dosage. It has strong irritation, and can cause pain by intramuscular or intravenous injection, and large vein should be selected for high incidence of local phlebitis. Midazolam is a benzodiazepine drug with the strongest water solubility, has the effect of 2-3 times of diazepam, has quick response, short duration and relatively quick waking, is suitable for treating patients with acute restlessness, can cause respiratory depression, blood pressure reduction and hypovolemia at a large dose, prolongs the accumulation and the sedation effect after long-term application, and has particularly obvious effect on patients with renal failure and tolerance phenomenon on part of patients. When applied in large dose, it has certain effect in inhibiting respiration, and can be used for treating apnea.

Phenothiazines belong to potent sedatives and can be used for the treatment of psychotic patients due to their anxiolytic properties. Chlorpromazine and promethazine are commonly used. The chlorpromazine is also called as hibernating, is commonly used for stabilizing and calming patients and is also used for mental patients. The medicine has strong autonomic nerve inhibiting effect, and a-adrenergic receptor blocking effect, and has effects of dilating blood vessel, lowering blood pressure, and easily causing orthostatic hypotension promethazine, has strong antihistaminic effect, can be applied in large dosage, has less influence on hemodynamics than chlorpromazine, is mainly used for administration before anesthesia and treating allergic diseases, and also has good tranquilizing and antiemetic effects,

the mechanism of action of butyrophenones is similar to that of phenothiazines. The antipsychotic action and extrapyramidal reaction are both strong, and the sedative and antihypertensive actions are weak. Because of its remarkable antimanic, antimigraine and delusional effects, it is used to treat schizophrenia and mania mainly manifested by agitation, hallucination and delusion, and has strong antiemetic effect, and can be used to treat emesis caused by various diseases and drugs, and also has therapeutic effect on persistent hiccup. Extrapyramidal system response is as high as 80%, common acute dystonia and akathisia can not be seen, and a large amount of long-term application can cause myocardial injury. The commonly used drugs include haloperidol and haloperidol. Wherein haloperidol is thought to be manic. The treatment effect of hallucinations and delusions is 50 times stronger than that of chlorpromazine, and is mainly used for psychotherapy. The haloperidol has stronger sedative effect than haloperidol, has less extrapyramidal symptoms, has better sedative and antiemetic effects, and is suitable for sedation of ICU patients, general anesthesia induction medicines and auxiliary medicines for nerve block anesthesia.

In summary, the conventional sedatives generally have obvious side effects, and care should be taken in selecting and administering the sedatives. Under such circumstances, if a drug with less side effects and stronger sedative effect can be developed, it is expected to improve the safety of administration and expand the range of choice of sedative drugs.

Lamivudine is a nucleoside analogue, and existing researches only find that lamivudine has competitive inhibition effect on synthesis and extension of virus DNA chains, and further is used as an antiviral drug: domperidone is an imidazole compound that acts as a peripheral dopamine receptor antagonist and exhibits gastrokinetic effects by blocking gastrointestinal dopamine receptors. In the prior art, lamivudine or domperidone has no sedative effect or central nervous system inhibiting effect on human bodies.

Disclosure of Invention

The invention aims to provide the application of dry preparation of a sedative drug by combining lamivudine and domperidone aiming at the technical defects of the prior art so as to expand the selectable range of the sedative drug.

Another technical problem to be solved by the present invention is that the indication ranges of lamivudine and domperidone in the prior art are limited.

The invention further solves the technical problem of how to further improve the sedative effect of the medicines.

In order to realize the technical scheme, the invention adopts the following technical scheme:

the application of the combination of lamivudine and domperidone in preparing sedative drugs.

Preferably, the mass ratio of lamivudine to domperidone is 3.18: 1.

Preferably, the medicine also comprises sodium hyaluronate, diosmin and nicotinamide.

Preferably, the medicine comprises the following components in parts by weight: 31.8 parts of lamivudine, 10 parts of domperidone, 6 parts of sodium hyaluronate, 1.5 parts of diosmin and 0.8 part of nicotinamide.

Preferably, in the medicament, the effective dose of lamivudine is 19.08 mg/d.

Preferably, the effective dose of domperidone in the medicine is 6 mg/d.

Preferably, the drug promotes GABA binding to its receptor and increases CT channel opening frequency.

Preferably, the medicament down-regulates the glutamate content in the cerebral cortex and hypothalamus.

Preferably, the drug agonizes the 5-hydroxytryptamine 1A receptor of the postsynaptic membrane.

Preferably, the dosage form of the medicament is oral preparation.

In the above technical scheme, GABA_AThe receptor is a supermolecular mass protein complex, and is a pentagon composed of 5 subunits embedded in the double lipid layer of nerve cell membraneA heterogeneous polypeptide oligomer having a GABA-gated chloride channel formed in its central portion. GABA_AThe receptor is the main inhibitory receptor in the central nervous system, its ligand and GABA_AThe corresponding sites of the receptor are combined, the conformation of the complex receptor is changed, and finally, the chloride ion flux in the chloride ion channel is regulated, so that the efficiency of GABA acting on the receptor is enhanced or weakened. There are two main types of GABA in the brain_AReceptor subtypes: omega₁Receptors are involved in sedative hypnosis: omega₂The receptors are involved in cognition, memory and psychomotor activity, and have muscle relaxant and anticonvulsant effects. The two receptors act together to achieve the sedative and hypnotic effects. The invention is based on experimental means to find that when the lamivudine and the domperidone are combined at a specific ratio, the GABA and the GABA can be promoted_AReceptor binding increases the opening frequency of chloride ion channels, thereby enhancing GABA-mediated chloride ion influx and further generating a neuroinhibition effect.

Glutamate is an excitatory amino acid, is widely distributed in the center, has relatively high levels in cerebral cortex and dorsal part of marrow, is a main excitatory transmitter, and plays an excitatory role in all neurons of the central nervous system to discharge the neurons. The medicine has high affinity with an N-methyl-D-aspartate receptor and long combination time, causes repeated development of the receptor channel, causes calcium ion inflow and intracellular calcium ion release caused by the activation of a metabotropic glutamate receptor, and continuously increases the concentration of the intracellular calcium ions, thereby causing agitation and excitation expression.

A large number of experiments and clinical researches show that the psychogenic hyperexcitability symptom is possibly related to the insufficient content of the 5-hydroxytryptamine, and when the 5-hydroxytryptamine 1A receptor of the presynaptic membrane is excited, the synthesis and the release of the 5-hydroxytryptamine are inhibited, so that the content of the 5-hydroxytryptamine in the synaptic cleft is obviously reduced. The drug combination scheme of the invention can be selectively combined with 5-hydroxytryptamine 1A receptors with high affinity, and has an exciting effect on the 5-hydroxytryptamine 1A receptors of the postsynaptic membranes, and inhibits the uptake of 5-hydroxytryptamine by synapses, so that the content of 5-hydroxytryptamine is up-regulated, and the sedative effect is achieved.

The pharmaceutical scheme provided by the invention has the advantages that the effective dose of lamivudine is 19.08mg/d, the effective dose of domperidone is 6mg/d, the effective doses are lower than the effective doses of the lamivudine and the domperidone which are used for treating the previous indications respectively, the safety is good under the dosage condition, and the safety can be proved based on the existing pharmacokinetic study.

The invention provides application of combination of lamivudine and domperidone in preparing a sedative drug. The technical scheme firstly screens compounds having potential influence on central nerves from existing medicines, and the result shows that although lamivudine and domperidone do not show a nerve inhibition effect when being singly administered, the lamivudine and the domperidone can play a definite sedative effect when being combined according to a specific ratio; moreover, its sedative effect is relatively mild and does not cause loss of communication ability or difficulty in arousal. On the basis, the invention researches the pharmacological properties of the medicament, and experiments show that the combined scheme of the invention can promote GABA and GABA_AReceptor binding, increasing the frequency of chloride channel opening; simultaneously, the glutamic acid content in the cerebral cortex and the hypothalamus of the experimental animal is reduced, so that the excitable neurotoxicity in the brain is reduced; moreover, the inhibitor can be selectively combined with a 5-hydroxytryptamine 1A receptor to inhibit the synaptic uptake of the 5-hydroxytryptamine, thereby up-regulating the content of the 5-hydroxytryptamine. Based on the discovery of the above new properties of the medicament, the invention determines the new application of the combination of lamivudine and domperidone in preparing the sedative, not only expands the selectable range of the sedative, but also improves the sedative effect to a certain extent.

Detailed Description

Hereinafter, specific embodiments of the present invention will be described in detail. Well-known structures or functions may not be described in detail in the following embodiments in order to avoid unnecessarily obscuring the details. Approximating language, as used herein in the following examples, may be applied to identify quantitative representations that could permissibly vary in number without resulting in a change in the basic function. Unless defined otherwise, technical and scientific terms used in the following examples have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs,

example 1

A sedative drug comprises lamivudine and domperidone, wherein the dosage of the lamivudine is 19.08mg/d, and the dosage of the domperidone is 6 mg/d.

Example 2

A sedative drug comprises lamivudine and domperidone, wherein the dosage of the lamivudine is 20.08mg/d, and the dosage of the domperidone is 5 mg/d.

Example 3

A sedative drug comprises lamivudine and domperidone, wherein the dosage of the lamivudine is 18.08mg/d, and the dosage of the domperidone is 7 mg/d.

Example 4

A sedative drug comprises lamivudine, domperidone, sodium hyaluronate, diosmin and nicotinamide, wherein the dosage of lamivudine is 19.08mg/d, the dosage of domperidone is 6mg/d, the dosage of sodium hyaluronate is 3.6mg/d, the dosage of diosmin is 0.9mg/d, and the dosage of nicotinamide is 0.48 mg/d.

Comparative example 1

Lamivudine was used as a sedative at a dose of 19.08 mg/d.

Comparative example 2

Domperidone is used as a sedative, and the dosage of the domperidone is 6 mg/d.

Comparative example 3

A sedative drug comprises lamivudine and domperidone, wherein the dosage of the lamivudine is 300mg/d, and the dosage of the domperidone is 30 mg/d.

Comparative example 4

Phenobarbital in a dose of 180 mg/d.

Comparative example 5

Diazepam in a dose of 40 mg/d.

Sedation experiment

Selecting score of 6 points according to Riker sedation and agitation scoring standard (Table 1)The above patients were used as subjects, 1000 subjects were randomly divided into 10 groups of 100 persons each, one group was blank control, and the other nine groups were administered according to the protocols of examples 1 to 4 and comparative examples 1 to 5, and scored again 2 hours after administration₁The experimental results are shown in table 2 below.

TABLE 1 Riker sedation and agitation scoring criteria

TABLE 2 results of sedation experiments characterized by Riker criteria

Grouping \ score values	7 points of	6 minutes	5 points of	4 is divided into	3 points of	2 is divided into	1 minute (1)
								Blank control	42	51	4	3	0	0	0
Example 1	0	0	1	83	14	2	0
								Example 2	0	13	32	48	7	0	0
Example 3	1	18	45	30	6	0	0
								Example 4	0	0	0	95	5	0	0
Comparative example 1	32	64	2	2	0	0	0
								Comparative example 2	45	52	1	2	0	0	0
Comparative example 3	47	44	8	1	0	0	0
								Comparative example 4	0	0	21	22	37	12	8
Comparative example 5	0	0	12	31	35	20	2

According to the RASS sedation degree evaluation table (Table 3), patients with score of +4 were selected as subjects, 1000 subjects were randomly divided into 10 groups of 100 subjects each, one group was blank-controlled, the remaining groups were administered according to the protocols of examples 1-4 and comparative examples 1-5 above, and scored again after 2h administration, with the experimental results shown in Table 4 below.

TABLE 3 RASS sedation assessment Table

TABLE 4 results of sedation experiments characterized by RASS criteria

Grouping \ score values	+4	+3	+2	+1	0	-1	-2	-3	-4	-5
											Blank control	92	4	3	1	0	0	0	0	0	0
Example 1	0	0	0	7	88	5	0	0	0	0
											Example 2	7	13	22	15	31	8	4	0	0	0
Example 3	10	15	19	18	28	7	3	0	0	0
											Example 4	0	0	0	2	95	3	0	0	0	0
Comparative example 1	90	8	2	0	0	0	0	0	0	0
											Comparative example 2	95	5	0	0	0	0	0	0	0	0
Comparative example 3	91	6	3	0	0	0	0	0	0	0
											Comparative example 4	0	0	7	26	34	15	8	4	4	2
Comparative example 5	0	2	12	11	25	13	12	14	6	5

From the above tables 2 and 4, it can be seen that when lamivudine and domperidone are used in combination at the doses of 19.08mg/d and 6mg/d, respectively (example 1), good sedation is exhibited, and the sedation is mild, and the subject's level of sedation is concentrated in a quiet, awake state without excessive sedation or coma. In contrast, phenobarbital (comparative example 4) or diazepam (comparative example 5) administered at therapeutic doses, although also producing a sedative effect, did not provide a concentrated level of sedation, with some patients having less sedative effects and some subjects showing over-sedation and coma. On the basis of the combination scheme of the invention, although the administration ratio is adjusted in a small range (examples 2 and 3), the therapeutic effect is shown to be certain, but even if the ratio is changed in such a small range, the therapeutic effect is obviously reduced compared with that in example 1. In addition, on the basis that lamivudine and domperidone are respectively combined at the doses of 19.08mg/d and 6mg/d, a specific amount of sodium hyaluronate, diosmin and nicotinamide (example 4) are further introduced to further improve the sedative effect, so that the sodium hyaluronate, diosmin and nicotinamide can be used as an improvement scheme of the technical scheme of the invention, comparative examples 1-3 do not show any sedative effect, and prove that the sedative effect is achieved only when the combined scheme of the invention is adopted, and no sedative effect is achieved when the single administration or other proportioning schemes are adopted. The results of the above experiments in tables 2 and 4 are mutually corroborated.

Pharmacological experiments

1. Effect of the inventive Agents on the binding of GABA to its receptors

Collecting male Kunming mouse, weighing 20-25g, performing intragastric administration, killing by pulling neck after 2h administration, rapidly separating four brain areas of hypothalamus, cerebral cortex, hippocampus and cerebellum, weighing, electrically homogenizing for 30 times in a homogenizer containing 2mL sucrose, centrifuging at 3000rpm for 10min, collecting supernatant, dividing into two tubes, centrifuging at 4 deg.C at 12000rpm for 20min, collecting precipitate, adding appropriate amount of 50mM Tris-HCL buffer solution, and repeating high speed centrifugation for two times to obtain GABA-welded product_AWhen the sediment of the coating preparation of the receptor is frozen and stored at the temperature of minus 20 ℃ for more than 16 hours and unfrozen, 100 mu L of 0.02 percent Tritom-100 is added into water bath at the temperature of 37 ℃ for 30 minutes, a proper amount of Tris-HCL buffer solution is added, high-speed centrifugation is repeated twice to fully remove endogenous inhibitors, and a proper amount of Tris-HCL buffer solution is added for suspension for standby application.

0.9mL of meningeal receptor suspension was added with 100. mu.L of 0.1. mu.M³Mixing H-GABA, and allowing binding reaction at room temperature overnight to allow receptor to react with GABA³H-GABA binds well to the equilibrium. The reaction was terminated by centrifugation, and the supernatant was discarded and washed twice with triple-distilled water. Adding 100 μ L of mixed solution of formic acid and hydrogen peroxide (3: 1) into the precipitate, and placing in an oven at 80 deg.C for digestion for 30 min. After cooling, 2mL of toluene scintillation fluid and 1mL of hexanediol ether were added to clarify the fluid and the dpm of the sample was measured on a liquid scintillation counter.

The protein content in the meningeal preparation was determined by Coomassie Brilliant blue method. The OD595 value of the sample was measured with a type 721 spectrophotometer using BSA as a standard upper line. According to the dpm value and the protein content, calculate and combine³I1-GABGABA of A_AReceptor concentration. The experimental result shows that after the medicine is administered, experimental objects GABA and GABA_AThe binding to the receptor is significantly enhanced, and it is reasonable to assume that the GABA-mediated chloride ion influx is correspondingly enhanced, thereby resulting in an inhibitory effect.

2. Influence of the drug of the invention on the content of glutamic acid in brain tissue

The rats are fasted for 12 hours before operation and water is forbidden for 4 hours. 10% chloral hydrate (0.30mL/100g) is used for intraperitoneal injection for anesthesia, and spontaneous breathing is ensured during the operation. Lying-down fixation, neck unhairing, strong iodine disinfection, incision along the middle of neck, separating bilateral common carotid artery, protecting vagus nerve, and tying near and far ends of bilateral common carotid artery with double heavy lines, and cutting off from the middle to ensure blocking of arterial direct current. Sterile operation is performed during operation, 2mL of gentamicin injection is given to an operation incision, and the incision is sutured. The animals were then transferred to a well ventilated animal house for feeding. After anaesthetizing the red rats, only bilateral common carotid arteries are separated, but the arteries are not ligated and separated. And (3) after operation, beginning intragastric administration at 3d, and evaluating the influence of the medicament on the content of glutamic acid in the brain tissue of the experimental animal after administration for 15 d.

Cutting off head rapidly, peeling off Hippocampus from both sides at low temperature, collecting in 1.5mL centrifugal tube, marking, and storing in-80 deg.C refrigerator for use. Taking about 50mg of hippocampus, adding 200 mu L of absolute ethyl alcohol, grinding into homogenate on an ice bench, sucking 200 mu L of homogenate, centrifuging for 20min at 16000r/min and 4 ℃, and taking 80 mu L of supernatant liquid to measure the content of glutamic acid by HPLC. The experimental results show that compared with the blank control, the glutamic acid content in the brain tissue of the rat under the condition of continuous administration is reduced by more than 30 percent.

3. Influence of the drug of the invention on the 5-hydroxytryptamine content of brain tissue

The method comprises the steps of collecting male Kunming mice, weighing 20-25g, carrying out intragastric administration, carrying out neck pulling for killing after 2h of administration, quickly separating four brain areas of a hypothalamus, a cerebral cortex, a hippocampus and a cerebellum, weighing, placing into a homogenizer filled with 2mL of cane sugar for electrically homogenizing for 30 times, centrifuging at the rotating speed of 3000rpm for 10min, taking supernate, and measuring the content of 5-hydroxytryptamine in a sample by adopting HPLC (high performance liquid chromatography), wherein a chromatographic column adopted is a Bondapak C18 stainless steel column (5 mu m, 250mm × 4.6.6 mm), and a mobile phase comprises methanol-water-acetic acid (70: 30: 0.1), the flow rate is 1mL/min, a fluorescence monitor is used, the excitation wavelength is 278nm, and the emission wavelength is 333 nm.

In conclusion, the medicine can exactly promote GABA and GABA_AReceptor binding, thereby increasing the chloride channel opening frequency; simultaneously, the content of glutamic acid in brain tissues of experimental animals can be reduced, so that the toxicity of excitant in the brain is reduced; moreover, the content of 5-hydroxytryptamine can be effectively adjusted. The pharmacological process is probably the principle of the sedative effect of the drug of the invention, thus providing sufficient pharmacological support for the sedative effect of the invention.

The embodiments of the present invention have been described in detail, but the description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention. Any modification, equivalent replacement, and improvement made within the scope of the application of the present invention should be included in the protection scope of the present invention.

Claims

1. The application of the combination of lamivudine and domperidone in preparing sedative drugs.

2. Use according to claim 1, wherein the mass ratio of lamivudine to domperidone is 3.18: 1.

3. The use according to claim 2, wherein said medicament further comprises sodium hyaluronate, diosmin, niacinamide.

4. The use according to claim 3, wherein the medicament comprises the following components in parts by weight: 31.8 parts of lamivudine, 10 parts of domperidone, 6 parts of sodium hyaluronate, 15 parts of diosmin and 0.8 part of nicotinamide.

5. The use according to claim 4, wherein the effective dose of lamivudine is 19.08mg/d in the medicament.

6. The use according to claim 5, wherein the effective amount of domperidone is 6 mg/d.

7. The use of claim 2, wherein the medicament promotes GABA binding to its receptor and increases Cl^-Channel open frequency.

8. The use of claim 2, wherein the medicament down-regulates glutamate content in the cerebral cortex and hypothalamus.

9. The use of claim 2, wherein the medicament agonizes the 5-hydroxytryptamine 1A receptor of the postsynaptic membrane.

10. The use according to claim 2, wherein the medicament is in the form of an oral dosage form.