Isoindoline-1, 3-dione derivatives, preparation method and application thereof, and pharmaceutical composition containing derivativesTechnical Field
The invention belongs to the field of pharmacy, and particularly relates to isoindoline-1, 3-dione derivatives, a preparation method thereof, application thereof and a pharmaceutical composition containing the derivatives.
Background
Pain is an unpleasant emotion with fear, tension, anxiety, etc. and with a painful sensation of cardiovascular, respiratory changes, which is usually accompanied by substantial or potential tissue damage. Currently marketed analgesics include non-steroidal anti-inflammatory drugs, opioid receptor agonists, anticonvulsants, and the like. Among them, opioid receptor agonists are the most effective drugs for treating pain, and have been widely used since several centuries. There are four subtypes of opioid receptors, namely Mu Opioid Receptor (MOR), delta Opioid Receptor (DOR), kappa Opioid Receptor (KOR), and opioid receptor-like receptor 1 (ORL-1 receptor). Wherein mu, delta and kappa opioid receptors all mediate analgesia, but have different side effects. Mu opioid-mediated respiratory depression, sedation, euphoria, nausea, urinary retention, bile cramping and constipation; activation of kappa opioid receptors produces a restless, aversive mood and has sedative and diuretic effects; delta opioid receptors can mediate the generation of rewards, respiratory depression, and convulsions.
Although opioid receptor agonists have potent analgesic effects, they are used with serious side effects, including strong respiratory depression (life threatening in severe cases), gastrointestinal dysfunction (e.g., nausea, vomiting, constipation), itching due to histamine release, and the like. Clinically used opioid receptor agonists are mostly used for exciting the mu opioid receptor, and activation of the mu opioid receptor can produce the strongest analgesic effect, but is accompanied by the most serious side effects.
In recent years, researchers have proposed a number of strategies to address the side effects of opioid agonists, but the incidence of common side effects of drugs designed and developed therefrom has remained high in preclinical and clinical studies. There is a need to develop an opioid receptor agonist with little side effects.
Disclosure of Invention
The invention aims to provide isoindoline-1, 3-dione derivatives, a preparation method and application thereof, and a pharmaceutical composition containing the derivatives.
The invention provides an application of a compound, pharmaceutically acceptable salt or optical isomer thereof in preparing delta opioid receptor agonist, wherein the structure of the compound is shown as a formula I:
I is a kind of
Wherein a is selected from 1,2,3 or 4;
R1、R2 is independently selected from C1-3 alkyl which is unsubstituted or substituted by one or more than two R3, or R1 and R2 are connected to form 5-6 membered saturated nitrogen heterocyclic group which is unsubstituted or substituted by one or more than two R4; the R3 groups are each independently selected from phenyl, halogenated phenyl, C1-2 alkoxy, halogenated C1-2 alkoxy, C1-2 alkyl, halogenated C1-2 alkyl, and the R4 groups are each independently selected from phenyl, halogenated phenyl, C1-2 alkoxy, halogenated C1-2 alkoxy, C1-2 alkyl, halogenated C1-2 alkyl.
Further, the delta opioid receptor agonist is an analgesic drug.
Further, in the compound, a is selected from 2 or 4;
R1、R2 is independently selected from C1-3 alkyl which is unsubstituted or substituted by one or more than two R3, or R1 and R2 are connected to form 5-6 membered saturated nitrogen heterocyclic group which is unsubstituted or substituted by one or more than two R4; the R3 groups are each independently selected from phenyl, C1-2 alkoxy, C1-2 alkyl, and the R4 groups are each independently selected from phenyl, C1-2 alkoxy, C1-2 alkyl.
Further, the structure of the compound is selected from one of the following structures:
。
Further, the pharmaceutically acceptable salt is hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoric acid, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate.
The invention also provides a compound, a pharmaceutically acceptable salt or an optical isomer thereof, wherein the structure of the compound is selected from one of the following structures:
。
Further, the pharmaceutically acceptable salt is hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoric acid, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate.
The invention also provides a pharmaceutical composition with delta opioid receptor agonism, which is a preparation prepared by taking compound 18 or compound 20, pharmaceutically acceptable salts or optical isomers thereof as active ingredients and adding pharmaceutically acceptable auxiliary materials.
The present invention also provides a process for preparing compound 18 or compound 20, comprising the steps of:
Reacting the compound 18-1 with the compound 18-2 to obtain a compound 18; or compound 20-1 is reacted with compound 20-2 to give compound 20.
The present invention also provides a process for preparing a pharmaceutically acceptable salt of compound 18 or compound 20, comprising the steps of:
(i) Compound 18, compound 20 were prepared as described above;
(ii) Compound 18 and compound 20 are each reacted with an acid to provide a pharmaceutically acceptable salt of the compound.
Definition of terms used in connection with the present invention: unless otherwise indicated, the initial definitions provided for groups or terms herein apply to the groups or terms throughout the specification; for terms not specifically defined herein, the meanings that one skilled in the art can impart based on the disclosure and the context.
The minimum and maximum values of carbon atom content in the hydrocarbon group are indicated by a prefix, for example, the prefix Ca-b alkyl represents any alkyl group containing from "a" to "b" carbon atoms. For example, C1-3 alkyl refers to straight or branched chain alkyl groups containing 1 to 3 carbon atoms.
Halogen is fluorine, chlorine, bromine or iodine.
By "pharmaceutically acceptable" is meant that the carrier, vehicle, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients comprising the pharmaceutical dosage form, and physiologically compatible with the recipient.
"Salts" are acidic and/or basic salts formed with inorganic and/or organic acids and/or bases of a compound or an optical isomer thereof, and also include zwitterionic salts (inner salts) and also include quaternary ammonium salts, such as alkylammonium salts. These salts may be obtained directly in the final isolation and purification of the compounds. Or by mixing the compound, or an optical isomer thereof, with a predetermined amount of an acid or a base as appropriate (for example, equivalent). These salts may be obtained by precipitation in solution and collected by filtration, or recovered after evaporation of the solvent, or by lyophilization after reaction in an aqueous medium.
The pharmaceutically acceptable salt of the present invention may be the hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoric acid, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate salt of the compound.
The compound provided by the invention has an agonizing effect on delta opioid receptors, can play an analgesic effect in vivo, and remarkably reduces the incidence rate of side effects of common opioid medicines. The compound can be used as delta opioid receptor agonist and has wide application prospect in preparing analgesic drugs with low side effects.
The preparation method of the compound is simple, has mild conditions and is suitable for industrial production.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 is a graph showing the effect of compounds activating delta opioid receptors on cAMP inhibition.
FIG. 2 shows analgesic effects of representative compound 21 on a rat CFA pain model, with the abscissa B representing baseline; c represents after molding. * Represents P <0.05 compared to the model group.
Figure 3. Influence of representative compound 21 and positive compound fentanyl on gastrointestinal mobility. * P <0.001 compared to the blank solvent group.
Detailed Description
The raw materials and equipment used in the invention are all known products and are obtained by purchasing commercial products.
The operation of the examples of the present invention was carried out at room temperature, which was 20-35 c, unless otherwise specified.
The compounds of the present invention were synthesized according to the following general synthetic route:
The following are examples of synthesis of specific compounds.
Example 1: preparation of Compounds 1-28 and pharmaceutically acceptable salts thereof
1. Preparation of Compound 7 and pharmaceutically acceptable salts thereof
2- (2-Bromomethyl) isoindole-1, 3-dione (0.508 g,1 eq) was dissolved in 4 mL DMF, to which were added K2CO3 (0.553 g,2 eq) and raw dibenzylamine (0.592 g,1.5 eq) in this order, and allowed to stand at room temperature for reaction for 16 hours. TLC monitoring reaction is complete, and decompression concentration is carried out to remove solvent; dissolving the residue in ethyl acetate, extracting with water, collecting the organic phase, adding anhydrous sodium sulfate to the organic phase, drying, concentrating, and separating by column chromatography (eluent: PE/EA=5/1) to obtain 0.533: 0.533 g (target compound 7), as pale yellow oily substance, and yield 72%.1H-NMR (CDCl3) δ 7.80 (dd,J= 5.5, 3.0 Hz, 2H), 7.72 (dd,J= 5.4, 3.1 Hz, 2H), 7.22 (m, 4H), 7.11 (m, 6H), 3.78 (t,J= 5.8 Hz, 2H), 3.59 (s, 4H), 2.72 (t,J= 5.9 Hz, 2H); HRMS m/z calcd. for C24H22N2O2 [M+H]+371.1760, found 371.1777.
0.533 G of the above-obtained target compound 7 was dissolved in 15mL of ethyl acetate, and an ethyl acetate solution (2 mL) of oxalic acid dihydrate (0.265 g,1.05 eq) was added thereto, followed by stirring at room temperature for 2h. After the reaction, filtering, dissolving the filter cake in 10 mL ultrapure water, and freeze-drying overnight in a freeze dryer to obtain a light and fluffy white solid, namely the oxalate of the target compound 7.
2. Preparation of Compounds 1-6, 8-28 and pharmaceutically acceptable salts thereof
According to the synthetic route of the general formula, the compounds 1-6, 8-28 and pharmaceutically acceptable salts thereof are synthesized by referring to the reaction conditions when the compound 7 and pharmaceutically acceptable salts thereof are synthesized.
The yield of the compound 1-28 obtained by the synthesis method is 60% -90%, and the purity is more than 95%.
The structures of compounds 1-28 are shown below and the characterization data for compounds 1-28 are shown in Table 1.
。
TABLE 1 HRMS data for compounds
The beneficial effects of the present invention are demonstrated by the test examples below.
Test example one, test of agonistic Activity of Compounds on delta opioid receptors
Delta opioid receptors, when activated, cause a decrease in intracellular cyclic adenosine monophosphate (cAMP) levels. The effect of the compounds of the present invention on cAMP content of CHO cells stably expressing delta opioid receptors was tested by the following in vitro cell experiments to determine the agonistic activity of the compounds on delta opioid receptors.
1. Experimental materials
The main equipment is as follows: multifunctional enzyme-labeled instrument (BMG), micro 384 well plate (Greiner)
The main reagent comprises: F12K Medium (Gibco), fetal bovine serum (Corning), bleomycin (Sigma), hygromycin B (Sigma), cAMP detection kit (Cisbio)
Cell line: CHO cells stably expressing delta opioid receptor (GENESCRIPT)
2. Experimental method
(1) Cell culture
CHO cells stably expressing delta opioid receptor were cultured in F12K medium containing 10% fetal bovine serum, 200 μg/μl bleomycin, 100 μg/μl hygromycin B and passaged once a day.
(2) Detection of agonistic Activity of Compounds on delta opioid receptors
CHO cells stably expressing delta opioid receptors were digested and centrifuged, plated into a micro 384-well plate, 3000 (5 μl) per well. mu.L of each concentration (final concentration of 10. Mu.M, 1. Mu.M, 100 nM, 10nM, 1 nM, 100 pM, 10 pM, 1 pM) of test compound solution was added to each well and incubated 45 min at 37℃in the absence of light. After adding 5. Mu. L CAMP CRYPTATE solution and 5. Mu.L of cAMP d2 antibody to each well, incubating at room temperature in the absence of light for 1: 1h, detection was performed on a multifunctional microplate reader (fluorescence emission intensity: 665 nm/620 nm). The use of GraphPad Software to fit the half maximal effect concentration (EC50) and the maximal stimulation percentage (Emax) of each compound on delta opioid receptor agonism indicates that the smaller the value of EC50, the greater Emax indicates the lower concentration required to effect, the stronger the effect and the better the compound effect.
3. Experimental results
The effect of each compound on activation of delta opioid receptors to cause cAMP inhibition is shown in fig. 1, and the results of EC50 and Ema of each compound on delta opioid receptor agonistic activity are shown in table 2, with ADL5859 as a positive control.
TABLE 2 EC50 and Emax for the agonistic Activity of the various Compounds on delta opioid receptors
Note that: "ND": no signal detected; "+":500 nM.ltoreq.EC50<5000 nM;"++"：50 nM≤EC50＜500 nM;"+++"：EC50 <50 nM.
"ND": no signal detected; "*": emax <60; "**": emax is more than or equal to 60 and less than 90; "***": emax is not more than 90.
The test results above demonstrate that: the compound of the invention can effectively excite delta opioid receptors, thereby playing an analgesic effect. Wherein the effect of the compound 21 is better, EC50 of the compound 21 is less than 50 nM, and Emax is more than or equal to 90.
Test example two, evaluation of analgesic efficacy
1. Experimental method
SD rats (weight of 250-300 g) are adapted to the experimental environment for 3 days. The skin in the middle of the left foot sole of the rat was stimulated with electrons vonfrey, and the contraction reaction was observed, which was used as an index of pain reaction. First, the mechanical stimulus basal pain threshold of rats was measured, and rats meeting the group-entry criteria each subcutaneously injected with 0.1 mL Freund's complete adjuvant (CFA) on the bottom of the left foot to induce pain. The mechanical stimulus pain threshold of the left foot was measured after about 16 h and the eligible rats were randomized into model groups, 6 tested compound groups each, male and female halves.
All groups of compounds tested were administered by intraperitoneal injection, with a compound concentration of 60. Mu. Mol/kg and a solvent of DMSO: tween: water = 1:1:8, the injection volume is 0.3 mL/100g; the model group i.e. the intraperitoneal injection of blank solvent. The mechanical stimulus pain threshold of the left hind paw of the rat at 20 min, 40 min, 1h, 1.5 h, 2h, respectively, was recorded after dosing. After the experiment was completed, data analysis was performed using GraphPad software. The mechanical stimulus pain threshold was increased and a statistical difference from the mechanical stimulus pain threshold of the blank solvent group at the corresponding time point represents that the tested compound had analgesic effect.
2. Experimental results
The evaluation results of the analgesic effect of the representative compound 21 of the present invention on the rat CFA pain model are shown in fig. 2. The results show that: representative compound 21 can rapidly and remarkably improve the pain threshold of hind paw mechanical stimulation of rats after injection (the blank solvent group with the P <0.05 vs), and has excellent analgesic effect.
Test example three, evaluation of side effects
Opioids often induce side effects, such as constipation, in the course of exerting analgesic effects. The following experiments prove that the side effects of the compounds of the invention are significantly reduced.
1. Experimental method
ICR mice (weighing 20-30 g) were fasted without water withdrawal overnight before the start of the experiment. On the day of the experiment, the animals were randomly divided into a blank solvent (normal) group, a positive compound (fentanyl) group, and a test compound group, 6 animals each, and a male and female half.
Blank solvent or test compound solution was injected intraperitoneally 30 min a in advance, the administration volume was 0.1: 0.1 mL/10: 10 g, and the administration dose of test compound solution was 60. Mu. Mol/kg.30 After min, each mouse is irrigated with a gastrointestinal function marker of 0.3 mL% (prepared by repeatedly boiling water solution containing 5% gum arabic and 10% activated carbon powder and cooling); the mice were sacrificed by cervical dislocation after the gastrointestinal function marker was lavaged for 30 min, the mesentery was dissected, the pylorus of the mice was cut to the ileocecum, and the length (S/cm) from the pylorus to the front of the ink and the total length (L/cm) from the pylorus to the ileocecum in the small intestine of the mice were measured by laying them on a table. Gastrointestinal mobility can be found by the following formula:
gastrointestinal mobility (%) =gastrointestinal function marker push rate (%) =s/l×100%
The gastrointestinal mobility was less than 50% and the statistical difference from the blank solvent group suggests that the tested compounds may induce constipation while exerting analgesic effects.
2. Experimental results
The results of the effect of the representative compound 21 of the present invention and the positive compound fentanyl on gastrointestinal mobility are shown in figure 3. When the analgesic effect is exerted, the classical opioid (positive control fentanyl) causes strong inhibition of gastrointestinal motility (gastrointestinal mobility < 50%) and induces constipation; representative compound 21 of the present invention did not cause gastrointestinal mobility abnormalities (gastrointestinal mobility > 50%) and did not present significant constipation. The experimental results show that the compound can be used as an analgesic with low side effect.
In summary, the invention provides isoindoline-1, 3-dione derivatives, and a preparation method and application thereof. The compound provided by the invention has an agonizing effect on delta opioid receptors, can play an analgesic effect in vivo, and remarkably reduces the incidence rate of side effects of common opioid medicines. The compound of the invention is used as delta opioid receptor agonist and has wide application prospect in preparing analgesic drugs with low side effects.