CN107174586B

CN107174586B - Pharmaceutical composition with arundoin derivative as active ingredient and application thereof

Info

Publication number: CN107174586B
Application number: CN201710092931.2A
Authority: CN
Inventors: 陈纪军; 尹秀娟; 马云保; 黄晓燕; 耿长安; 张雪梅
Original assignee: Kunming Institute of Botany of CAS
Current assignee: Kunming Institute of Botany of CAS
Priority date: 2017-02-21
Filing date: 2017-02-21
Publication date: 2020-08-11
Anticipated expiration: 2037-02-21
Also published as: CN107174586A

Abstract

The invention provides a pharmaceutical composition taking arundoin derivatives (1-6) as active ingredients, a derivative and a pharmaceutical composition thereof which are used as melatonin and serotonin receptor agonists, and application thereof in preparing medicines for treating or improving central nervous system diseases related to melatonin and 5-serotonin receptors.

Description

Pharmaceutical composition with arundoin derivative as active ingredient and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition taking a graminine derivative shown in structural formulas 1-6 as an active ingredient, a preparation method thereof, a melatonin and 5-hydroxytryptamine receptor agonist serving as the derivative and the pharmaceutical composition thereof, and application of the derivative and the pharmaceutical composition thereof in preparation of medicines for treating or improving central nervous system diseases related to melatonin and serotonin receptors.

Background

Melatonin (Melatonin) receptor is a neuroendocrine hormone secreted by the pineal gland, controlled by the supranuclear hypothalamic cross, and regulates circadian rhythms, which are manifested by a peak at nightDaytime decreases to a trough value. McCord and Allan found in 1917 that the bovine pineal bodies can regulate the changes of Rana pipiens tadpole skin in the environment, and determined that the structure of melatonin is N-acetyl-5-methoxytryptamine. Melatonin is classified as MT according to melatonin function and pharmacological profile₁，MT₂And MT₃Three subtypes, these receptors are considered as potential pharmaceutically active targets. Wherein MT₃Receptor affinity sites are predominantly located in human homeosomal hamster Quinone Reductase (QR) enzyme with detoxifying properties₂) The above. Melatonin receptor subtype MT found in mammals₁And MT₂Currently, common melatonin receptor agonists are Tasimelteon (VEC-162), Ramelteon (TAK-375), β -methyl-6-chloromelatonin (LY-156735, TIK-301), Agomelatine (Agomelatine)^[11]Etc.; common melatonin receptor antagonists: n-acetyl-2-benzyltryptamine (Luzindole), 4-Phenyl-2-propionoaminotetralin (4-P-PDOT), and the like.

With the intensive research on natural products, more and more natural small molecules are attracting the interest of medicinal chemists. Natural small molecule arundoin (N, N-dimethylaminomethylindole), originally isolated from Arundo Donax (Arundo Donax L.) of the Arundo genus of asian grass (Gramineae) by Orekhov and Norkina, has a chemical structure similar to that of melatonin receptors and a broad range of biological activities: including vasoconstriction, antioxidation, neurotransmitter 5-HT_2AReceptor antagonists, etc. The arundoin can inhibit CVB3 host Cell Pathological Effect (CPE), and can be used as an anti-CVB 3 virus drug [ CN 105748474 ] by inhibiting early CVB3 virus replication in host cells]. Giantreed alkali or pharmaceutically acceptable salt as active ingredient has good skin whitening, elasticity, anti-wrinkling or skin moisturizing effects on human body [ KR 2016020240]。

At present, no report is found on the use of the arundoin derivatives (1-6) provided by the invention as melatonin and 5-hydroxytryptamine receptor agonists and medicaments for resisting mental diseases as active ingredients, and no report is found on the use of the derivatives or the pharmaceutical compositions thereof as melatonin and 5-hydroxytryptamine receptor agonists in the preparation or treatment of anti-mental disease medicaments.

Disclosure of Invention

The invention aims to provide a novel pharmaceutical composition taking the Giantreed alkali derivatives (1-6) with medicinal value as active ingredients, a preparation method thereof, application of the derivatives and the pharmaceutical composition thereof as melatonin and 5-hydroxytryptamine receptor agonists and application of the derivatives and the pharmaceutical composition thereof in preparing medicines for treating or improving central nervous system diseases related to melatonin and 5-hydroxytryptamine receptors.

In order to achieve the above purpose of the present invention, the present invention provides the following technical solutions:

the pharmaceutical composition contains the arundoin derivatives 1-6 shown in the following structural formula and a pharmaceutically acceptable carrier or excipient.

The invention also provides a method for preparing the arundoin derivatives 1-6 in the pharmaceutical composition, wherein the arundoin is respectively reacted with ethylamine, pyrrole, piperidine, N-isopropylpiperazine, N-methyl homopiperazine and homopiperazine in anhydrous toluene, the reaction is performed at the temperature of 110 ℃ under reflux until the reaction is finished, the solvent is recovered under reduced pressure to prepare a crude product, and the crude product is subjected to silica gel column chromatography purification (diethylamine/methanol/chloroform, v/v/v, 2/4/94-3/5/92) to prepare target compounds 1-6.

The invention also provides application of the pharmaceutical composition in preparing melatonin and 5-hydroxytryptamine receptor agonists, and application of the pharmaceutical composition in preparing medicines for treating or improving central nervous system diseases related to melatonin and 5-hydroxytryptamine receptors.

The invention also provides application of the arundoin derivatives 1-6 shown in the following structural formula in preparing medicines for treating or improving central nervous system diseases related to melatonin and 5-hydroxytryptamine receptors, application of the arundoin derivatives 1-6 in preparing anti-mental disease medicines, and application of the arundoin derivatives 1-6 in preparing melatonin and 5-hydroxytryptamine receptor agonists.

Melatonin and 5-hydroxytryptamine receptor agonistic activity are used as guidance to discover that the arundoin derivatives 1-6 have the effect of resisting mental diseases. The invention obtains 6 derivatives through the structural modification of the arundoin, and the derivatives have better agonistic activity, wherein the activity of the

compounds

2 and 6 is better, and the derivatives have better activity on a melatonin receptor MT₁EC of (1)₅₀Values of 0.51 and 0.50 mm respectively; for 5-HT_1AEC of (1)₅₀The values were 0.28 and 0.23 mm respectively.

When the compound of the present invention is used as a medicament, it may be used as it is or in the form of a pharmaceutical composition. The pharmaceutical composition contains 0.1-99%, preferably 0.5-90%, of the compound of the present invention, the remainder being pharmaceutically acceptable, non-toxic and inert pharmaceutically acceptable carriers or excipients for humans and animals.

The pharmaceutically acceptable carrier or excipient is one or more of solid, semi-solid and liquid diluents, fillers and pharmaceutical adjuvants. The pharmaceutical composition of the present invention is used in the form of a dose per unit body weight. The medicine of the present invention may be administrated through injection and oral taking.

Description of the drawings:

FIG. 1 is a schematic structural diagram of the arundoin derivatives (1-6) of the present invention.

Detailed Description

In order to better understand the substance of the present invention, the preparation method and pharmacological effect results of the arundoin derivatives (1-6) of the present invention are illustrated by the examples of the present invention, but the present invention is not limited thereto.

In the following examples, high resolution electrospray ionization mass spectrometry (HRESIMS) was performed on an LCMS-IT-TOF mass spectrometer (Shimadzu, Kyoto, Japan)Fixed, nuclear magnetic resonance spectrum (¹H and¹³c NMR from Bruker AM 400(¹H/¹³C, 400MHz/100MHz) nuclear magnetic resonance apparatus (Bruker, Bremerhaven, Germany) using TMS (tetramethylsilane) as internal standard. Column chromatography silica gel (200-300 mesh) and thin-layer chromatography silica gel GF254 are all produced by Qingdao Meigaoji Co. The reagents were purchased from Alfa Aesar, carbofuran and Acros.

Compound preparation example 1:

dissolving arundoin (2mmol) and ethylamine, pyrrole, piperidine, N-isopropylpiperazine, N-methyl homopiperazine and homopiperazine (2-10 mmol) in 10mL of anhydrous toluene respectively, refluxing at 110 ℃ until the reaction is finished, recovering the solvent under reduced pressure to obtain a crude product, and purifying by silica gel column chromatography (diethylamine/methanol/chloroform, v/v/v, 2/4/94-3/5/92) to obtain a target compound (1-6):

compounds 1-6 structure determination data:

n, N-Diethyl-3-indolylmethylamine (N, N-Diethyl-3-indolylmethylamine, 1) was a white powder, and the yield was 97%.¹H NMR (400MHz,CDCl₃)_H:8.93(s,1H,NH),7.78‐7.00(m,5H,H‐2,4,5,6,7),3.87(s,2H,‐CH₂N),2.69‐2.65(m,4H,

N-Pyrrolyl-3-indolylmethylamine (2) was obtained in the form of a white powder with a yield of 96%.¹H NMR(400MHz, CDCl₃)_H:9.27(s,1H,NH),7.75‐7.00(m,5H,H‐2,4,5,6,7),3.91(s,2H,CH₂N),2.68(m,4H,H‐2',5'),

Piperazinyl-3-indolylmethylamine, 4) as a white powder in a yield of 93%.¹H NMR(400MHz,CDCl₃)_H:9.57(s,1H,NH), 7.73‐6.94(m,5H,H‐2,4,5,6,7),3.78(s,2H,CH₂N),2.69‐2.60(m,9H,H‐1”,2',3',5',6'),1.05(d,6H,Me).¹³C NMR(100MHz,CDCl₃):136.2(s,C‐8),128.3(s,C‐9),124.4(d,C‐2),121.7(d,C‐6),119.3(d,C‐5),119.3 (d,C‐4),111.3(s,C‐3),111.2(d,C‐7),54.5(d,C‐1”),53.1(t,CH₂N),53.1(t,C‐3',5'),48.7(t,C‐2',6'),18.7(q,

7'),2.17(s,1H,NH),1.82‐1.76(m,2H,H‐3'),.¹³C NMR(100MHz,CDCl₃):136.2 (s,C‐8),128.1(s,C‐9),123.8(d,C‐2),121.7(d,C‐6),119.3(d,C‐5),119.2(d,

3-indolymethyalamine, 6) yellow amorphous powder, yield 80%.¹H NMR(400MHz,

CDCl₃)_H:10.05(s,1H,NH),7.80‐7.02(m,5H,H‐2,4,5,6,7),3.89(s,2H,CH₂N),2.89‐2.85(m,4H,H‐2',4'), 2.75‐2.69(m,4H,H‐6',7'),2.40(s,3H,NMe),1.89‐1.85(m,2H,H‐3').¹³C NMR(100MHz,CDCl₃):136.4(s, C‐8),128.1(s,C‐9),124.3(d,C‐2),121.6(d,C‐6),119.4(d,C‐5),119.1(d,C‐4),112.6(s,C‐3),111.3(d,C‐7),57.5(t,C‐6'),56.8(t,C‐7'),54.3(t,C‐4'),54.0(t,C‐2'),53.6(t,CH₂N),46.9(q,NMe),27.0(t,C‐3').ESIMS: m/z 244[M+H]⁺,HRESIMS:C₁₅H₂₁N₃[M+H]⁺Found 244.1742, calculated 244.1808.

The pharmacological effects of the arundoin derivatives of the present invention are illustrated below by the melatonin and 5-hydroxytryptamine receptor agonistic activity pharmacological effects test:

test example 1:

the arundoin derivatives (1-6) prepared in the preparation examples of the compounds of the invention are applied to human renal epithelial cells MT₁-HEK 293 and 5-HT_1A-measurement of the agonist activity on the receptor in a model of HEK293 cells.

1 agonist Activity test section

1.1 materials and instruments

MT for Activity screening₁And 5-HT_1AThe cell lines correspond to human kidney epithelial cells MT respectively₁-HEK 293 and 5-HT_1AHEK293, cell culture Medium containing 10% fetal bovine serum (Dulbecco's Modified Eagle Medium, DMEM), 10% FBS, HBSS from GIBCO; melatonin (CAS: 73-31-4) was purchased from Damas-beta, Inc. (Basel, Switzerland); the positive control drugs agomelatine (CAS: 138112-76-2) and 5-hydroxytryptamine (CAS: 50-67-9) were purchased from Saen chemical technology (Shanghai) Inc. and Yunnan Zehao commercial Inc., respectively; the leave-in Calcium 8 Kit (Wash Free Fluo-8 Calcium Asaay Kit, HD 03-0010, HDB Biosciences Co. Ltd, Shanghai, China). CO2 constant temperature incubator Thermo Forma 3310 (USA); inverted biomicroscope model XD-101 (nanjing); flexstation 3 desktop multifunctional microplate reader (Molecular Devices, Calif., USA).

1.2 Experimental procedures

Preparing to obtain tropine esters of organic acids with different skeletons, and applying the tropine esters to human kidney epithelial cells MT₁-HEK 293 and 5-HT_1AMT of the synthesized derivatives on a model of HEK293 cells with agomelatine and 5-hydroxytryptamine as positive controls₁And 5-HT_1AAnd (3) measuring the receptor agonistic activity. MT (multiple terminal)₁And 5-HT_1ACompound pair MT for receptor agonistic activity₁And 5-HT_1AReceptor activation rate and half maximal effective concentration Expression (EC)₅₀)， EC₅₀The concentration at which the drug produces 50% of the maximal effect on the corresponding symptom. EC was performed using the basic Biometrics and mapping integration software GraphPad Prism 5.0 developed by GraphPadSoftware Inc₅₀References to computational, specific test methods^[17‐19]Cells were plated at a density of 4 × 104/well in Matrigel-coated 96-well black-walled transfixion plates in CO using Dulbecco's modified Eagle Medium₂Culturing in a constant temperature incubator at 37 ℃ with the concentration of 5% for 24 h. The supernatant was aspirated, the original medium was discarded, and 100. mu.l/well of freshly prepared dye solution was added, and the mixture was protected from light at 37 ℃ for 60 min. Preparing a sample to be tested: preparing samples to be tested with different concentrations, and placing the samples to be tested into the other transparent bottom plate. The two plates are simultaneously placed in a FlexStation 3 desktop multifunctional microplate reader. Experimental data from Flex Station 3Reading by multi-functional enzyme-linked immunosorbent assay (EC)₅₀Values were calculated by Graph Pad Prism 5 software, 100% agonism Δ a/Δ c × 100 (a: test sample; c: positive control).

2. As a result: the concentration is 1.00mM, and the agonistic activity of the arundoin and the derivatives (1-6) thereof is shown in Table (1):

TABLE 1 Giantrodia alkali derivatives vs. MT₁And 5-HT_1AAgonistic activity of receptors^a

Note:^aall tested compounds were at a concentration of about 1Mm, agomelatine at a test concentration of 1.11 μ M, and 5-hydroxytryptamine at a test concentration of 6.67 μ M.

The agonistic activity is expressed as the average of three determinations

(n＝3).

TABLE 2 half Effective Concentration (EC) of

derivatives

2 and 6₅₀,mM)

Note: the test concentration range of the compound 1-6 is 0.02-1.52 mu M;

average of three determinations, EC₅₀Is expressed as

(n＝3)。

3. And (4) conclusion: the experimental results show that determined Arundinine derivative pairs MT₁And 5-HT_1AThe receptor has stronger excitability activity and has potential regulation or treatment effects related to melatonin and the receptor.

Formulation example 1:

the obtained arundoin derivatives 1-6 prepared by the method of preparation example 1 are dissolved by a small amount of DMSO respectively or mixed, and then are added with water for injection by a conventional method, and then are subjected to fine filtration, encapsulation and sterilization to prepare injection.

Formulation example 2:

the preparation method of the preparation example 1 is that the arundoin derivatives 1-6 are prepared firstly, and are dissolved by a small amount of DMSO respectively or mixed, then the obtained solution is dissolved in sterile water for injection, stirred to be dissolved, filtered by a sterile suction filter funnel, then sterile fine filtered, subpackaged in ampoules, frozen and dried at low temperature, and then sterile melt-sealed to obtain the powder injection.

Formulation example 3:

the method of preparation example 1 is firstly used for preparing the arundoin derivatives 1-6, and the arundoin derivatives and the excipient are added into the mixture respectively or mixed according to the weight ratio of the arundoin derivatives to the excipient of 9:1 to prepare powder.

Formulation example 4:

the preparation method of the preparation example 1 is firstly used for preparing and obtaining the arundoin derivatives 1-6, and the arundoin derivatives are respectively or mixed, added with excipient according to the weight ratio of the arundoin derivatives to the excipient of 5:1, granulated and tabletted.

Formulation example 5:

the arundoin derivatives 1-6 obtained by the method of preparation example 1 were prepared first, and prepared into oral liquid by a conventional oral liquid preparation method, separately or mixed.

Formulation example 6:

the arundoin derivatives 1-6 obtained by the method of preparation example 1 are prepared, respectively or mixed, and the excipient is added according to the weight ratio of the arundoin derivatives to the excipient of 5:1 to prepare capsules.

Formulation example 7:

the preparation method of the preparation example 1 is firstly used for preparing the arundoin derivatives 1-6, and the arundoin derivatives and the excipients are respectively or mixed, added with the excipients according to the weight ratio of 3:1 and prepared into capsules.

Formulation example 8:

the arundoin derivatives 1-6 obtained by the method of preparation example 1 are prepared, respectively or mixed, and the excipient is added according to the weight ratio of the arundoin derivatives to the excipient of 5:1 to prepare granules.

Claims

1. The application of the pharmaceutical composition of the arundoin derivatives 2 and 6 shown in the following structural formula in the preparation of melatonin and 5-hydroxytryptamine receptor agonist,

2. the use of a pharmaceutical composition of arundoin derivatives 2 and 6 of the formula in the manufacture of a medicament for the treatment or amelioration of central nervous system disorders associated with melatonin and 5-hydroxytryptamine receptors,

3. the use of arundoin derivatives 2 and 6 of the formula in the manufacture of a medicament for the treatment or amelioration of central nervous system disorders associated with melatonin and 5-hydroxytryptamine receptors,

4. the use of the arundoin derivatives 2 and 6 of the following structural formula in the preparation of melatonin and 5-hydroxytryptamine receptor agonists,