CN107174586B - Pharmaceutical composition and application thereof using luteolin derivatives as active ingredients - Google Patents

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

Medicinal composition and application thereof using luteolin derivative as active ingredient

technical field

The invention belongs to the technical field of medicine, and in particular, relates to a pharmaceutical composition using the rhodophylline derivatives shown in structural formulas 1-6 as active ingredients, their preparation method, and the use of such derivatives and their pharmaceutical compositions as Melatonin and serotonin receptor agonists, and their use in preparing medicines for treating or improving central nervous system diseases related to melatonin and serotonin receptors.

Background technique

Melatonin receptor is a neuroendocrine hormone secreted by the pineal gland, which is controlled by the suprachiasmatic nucleus of the hypothalamus and can regulate the circadian rhythm, which is characterized by peaks at night and troughs during the day. McCord and Allan in 1917 discovered that the bovine pineal gland can regulate changes in the skin of the Rana pipien tadpole in the environment, and determined the structure of melatonin as N-acetyl-5-methoxytryptamine. Melatonin is divided into three subtypes, MT ₁ , MT ₂ and MT ₃ , according to its functional and pharmacological properties, and these receptors are regarded as potential drug-active targets. Among them, the MT ₃ receptor affinity site is mainly located on the quinone reductase (QR ₂ ) of the human homologous hamster with detoxification properties. The melatonin receptor subtypes MT ₁ and MT ₂ present in mammals have been cloned. The two receptor subtypes belong to the G-coupled protein receptor family and share some specific amino acid sequences, showing 60 at the amino acid level. % homology and unique pharmacological activity. At present, common melatonin receptor agonists: tasimelteon (Tasimelteon, VEC-162), ramelteon (Ramelteon, TAK-375), β-methyl-6-chloromelatonin (LY- 156735, TIK-301), Agomelatine ^[11] , etc.; common melatonin receptor antagonists: N-acetyl-2-benzyltryptamine (Luzindole), 4-Phenyl-2-propionamidotetralin (4-P-PDOT) etc.

With the in-depth study of natural products, more and more natural small molecules have attracted the interest of medicinal chemists. The natural small molecule rhodophylline (N,N-dimethylaminomethyl indole) was originally isolated from the Asian Gramineae plant Arundo Donax L. by Orekhov and Norkina. Similar chemical structure and broad biological activity of melatonin receptor: including vasoconstrictor, antioxidant, antagonist of neurotransmitter 5-HT _2A receptor, etc. Lubaline can inhibit the CVB3 host cytopathic effect (CPE), by inhibiting the early CVB3 virus replication in host cells, as an anti-CVB3 virus drug [CN 105748474]. Lubaline or pharmaceutically acceptable salt as an active ingredient has good skin whitening, elasticity, anti-wrinkle or skin moisturizing effect on human body [KR 2016020240].

At present, there is no report on the rutabine derivatives (1-6) provided by the present invention as melatonin and serotonin receptor agonists and as medicines for antipsychotic active ingredients. Melatonin and serotonin receptor agonists have not been reported in the preparation or treatment of antipsychotic drugs.

SUMMARY OF THE INVENTION

The present invention aims to provide a new pharmaceutical composition with medicinal value rudoline derivatives (1-6) as active ingredients, a preparation method thereof, and the derivatives and pharmaceutical compositions thereof as melatonin and serotonin receptor agonists, and their use in the manufacture of a medicament for the treatment or amelioration of central nervous system diseases associated with melatonin and serotonin receptors.

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

A pharmaceutical composition, comprising the rutabine derivatives 1-6 represented by the following structural formulas, and a pharmaceutically acceptable carrier or excipient.

At the same time, the present invention provides a method for preparing the rudolidine derivatives 1 to 6 in the pharmaceutical composition, wherein rudarine is respectively mixed with ethylamine, pyrrole, piperidine, N-isopropylpiperazine, N-methyl homogenate Piperazine and homopiperazine were reacted in anhydrous toluene, refluxed at 110 ° C until the end of the reaction, and the solvent was recovered under reduced pressure to prepare a crude product, which was purified by silica gel column chromatography (diethylamine/methanol/chloroform, v/v/v , 2/4/94～3/5/92) to prepare the target compound 1-6.

The present invention also provides the application of the pharmaceutical composition in the preparation of melatonin and serotonin receptor agonists, and the preparation of the pharmaceutical composition in the preparation of treatment or improvement with melatonin and serotonin receptors The use of medicines related to diseases of the central nervous system.

The present invention also provides the use of the rutabine derivatives 1-6 represented by the following structural formulas in the preparation of medicaments for treating or improving the diseases of the central nervous system related to melatonin and serotonin receptors, as well as the application of lucidum The application of base derivatives 1-6 in the preparation of antipsychotic drugs, and the application of rutabine derivatives 1-6 in the preparation of melatonin and serotonin receptor agonists.

Guided by the agonistic activities of melatonin and serotonin receptors, rubarine derivatives 1-6 were found to have antipsychotic effects. The present invention obtains 6 derivatives by modifying the structure of luteolin and has better agonistic activity, wherein compounds 2 and 6 have better activities, and the EC ₅₀ values for melatonin receptor MT ₁ are 0.51 and 0.50 mM respectively; The _EC50 values for 5-HT _1A were 0.28 and 0.23 mM, respectively.

When the compound of the present invention is used as a medicine, it can be used directly 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, and the rest are 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 solid, semi-solid and liquid diluents, fillers and pharmaceutical preparation adjuvants. The pharmaceutical composition of the present invention is used in a dosage per body weight. The medicament of the present invention can be administered by injection (intravenous injection, intramuscular injection) and oral administration.

Description of drawings:

Fig. 1 is the structural schematic diagram of rhodophylline derivatives (1-6) of the present invention.

Detailed ways

In order to better understand the essential content of the present invention, the examples of the present invention will be used to illustrate the preparation method and pharmacological effect results of the fenugreek derivatives (1-6) of the present invention, but the present invention is not limited by this.

In the following examples, high-resolution electrospray ionization mass spectrometry (HRESIMS) was measured on a LCMS-IT-TOF mass spectrometer (Shimadzu, Kyoto, Japan), and nuclear magnetic resonance spectra ( ¹ H and ¹³ C NMR) were obtained by Bruker AM 400 ( ¹ H / ¹³ C, 400MHz/100MHz) nuclear magnetic resonance apparatus (Bruker, Bremerhaven, Germany), with TMS (tetramethylsilane) as the internal standard. Column chromatography silica gel (200-300 mesh) and thin layer chromatography silica gel GF254 are produced by Qingdao Meigao Group Co., Ltd. Reagents were purchased from Alfa Aesar, Bailingwei and Acros.

Compound Preparation Example 1:

Lubaline (2mmol) and ethylamine, pyrrole, piperidine, N-isopropylpiperazine, N-methylhomopiperazine and homopiperazine (2-10mmol) were dissolved in 10mL of anhydrous toluene, refluxed at 110℃ To the end of the reaction, the solvent was recovered under reduced pressure to prepare a crude product, which was purified by silica gel column chromatography (diethylamine/methanol/chloroform, v/v/v, 2/4/94～3/5/92) to obtain the target compound ( 1‐6):

Structure determination data for compounds 1-6:

N,N-Diethyl-3-indolylmethylamine (N,N-Diethyl-3-indolylmethylamine, 1): white powder, yield 97%. ¹ H NMR (400 MHz, CDCl ₃ ) δ _H : 8.93 ( s,1H,NH),7.78-7.00(m,5H,H-2,4,5,6,7),3.87(s,2H,-CH ₂ N),2.69-2.65(m,4H,

Piperazinyl‐3‐indolylmethylamine,4):白色粉末,收率93％.¹H NMR(400MHz,CDCl₃)δ_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)δ: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)δ: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‐indolylmethylamine,6):黄色无定型粉末,收率80％.¹H NMR(400MHz,

CDCl₃)δ_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)δ: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]⁺测定值244.1742,计算值为244.1808.N-Pyrrolyl-3-indolylmethylamine, 2: white powder, yield 96%. ¹ H NMR (400 MHz, CDCl ₃ ) δ _H : 9.27(s, 1H, NH), 7.75 ‐7.00(m,5H,H‐2,4,5,6,7),3.91(s,2H,CH ₂ N),2.68(m,4H,H‐2',5'),

Piperazinyl‐3‐indolylmethylamine, 4): white powder, yield 93%. ¹ H NMR (400MHz, CDCl ₃ ) δ _H : 9.57(s, 1H, NH), 7.73‐6.94(m, 5H, H‐2, 4,5,6,7),3.78(s,2H,CH ₂ N),2.69‐2.60(m,9H,H‐1",2',3',5',6'),1.05(d, 6H,Me). ¹³ C NMR (100MHz, CDCl ₃ )δ: 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, CH ₂ N), 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'), . ¹³ C NMR (100MHz, CDCl ₃ )δ: 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‐indolylmethylamine, 6): yellow amorphous powder, yield 80%. ¹ H NMR (400MHz,

CDCl ₃ )δ _H : 10.05(s,1H,NH),7.80-7.02(m,5H,H-2,4,5,6,7),3.89(s,2H,CH ₂ N),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'). ¹³ C NMR (100MHz, CDCl ₃ )δ: 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,CH ₂ N),46.9(q,NMe),27.0(t,C‐3 ').ESIMS: m/z 244[M+H] ⁺ , HRESIMS: C ₁₅ H ₂₁ N ₃ [M+H] ⁺ found 244.1742, calculated 244.1808.

The pharmacological action results of the rubaline derivatives of the present invention are described below by the pharmacological action test of melatonin and serotonin receptor agonistic activity:

Test Example 1:

The rhodophylline derivatives (1-6) prepared in the above-mentioned compound preparation examples of the present invention were tested for their agonistic activity on receptors on human renal epithelial cell MT ₁ -HEK293 and 5-HT _1A -HEK293 cell models.

1 Agonistic activity experimental part

1.1 Materials and Instruments

The MT ₁ and 5-HT _1A cell lines used in the activity screening correspond to human renal epithelial cells MT ₁ -HEK293 and 5-HT _1A -HEK293, respectively, in cell culture medium (Dulbecco's Modified Eagle Medium, DMEM) containing 10% fetal bovine serum, 10% FBS, HBSS were purchased from GIBCO; Melatonin (CAS: 73-31-4) was purchased from Damas-beta (Basel, Switzerland); positive control drug agomelatine (CAS: 138112-76-2) and 5-hydroxytryptamine (CAS: 50-67-9) were purchased from San Chemical Technology (Shanghai) Co., Ltd. and Yunnan Zehao Trading Co., Ltd. respectively; Wash Free Fluo-8 Calcium Asaay Kit, HD03 ‐0010, HDB Biosciences Co. Ltd, Shanghai, China). CO2 constant temperature incubator Thermo Forma 3310 (USA); Inverted biological microscope XD-101 (Nanjing); Flexstation 3 desktop multi-plate reader (Molecular Devices, California, USA).

1.2 Experimental process

The organic acid tropine ester with different backbones was prepared, and the synthesized derivatives were tested on human renal epithelial cell MT ₁ -HEK293 and 5-HT _1A -HEK293 cell models, with agomelatine and serotonin as positive controls. MT ₁ and 5-HT _1A receptor agonistic activity assay. MT ₁ and 5-HT _1A receptor agonistic activity is expressed by compound on MT ₁ and 5-HT _1A receptor agonism rate and half effective concentration (EC ₅₀ ), EC ₅₀ - the concentration at which the drug produces 50% of the maximal effect on the corresponding symptom . EC ₅₀ was calculated by using ^GraphPad Prism 5.0, the basic biostatistics and drawing comprehensive software developed by GraphPad Software. Using Dulbecco's modified Eagle's medium, cells were cultured at a density of 4 × 104/well in Matrigel-coated 96-well black-walled permeable bottom plates in a 37°C incubator with a _CO concentration of 5% for 24h. Aspirate the supernatant, then discard the original medium, add 100 μl/well of freshly prepared dye solution, and protect from light at 37°C for 60 min. Prepare the sample to be tested: prepare the sample to be tested with different concentrations, and place the test sample into another transparent bottom plate. Both plates were placed in the FlexStation 3 benchtop multi-plate reader at the same time. The experimental data was read by Flex Station 3 desktop multi-function microplate reader, and the EC ₅₀ value was calculated by Graph Pad Prism 5 software; activation rate=Δδa/Δδc×100% (a: test sample; c: positive control).

2. Results: At a concentration of 1.00 mM, the agonistic activities of luteolin and its derivatives (1-6) are shown in Table (1):

Table 1 Agonistic activities of rubarine derivatives on MT ₁ and 5-HT _1A receptors ^a

为三次测定的平均值，激动活性表达为

(n＝3).Notes: ^a All tested compounds were at a concentration of approximately 1 Mm, agomelatine was tested at a concentration of 1.11 μM, and serotonin was tested at a concentration of 6.67 μM.

is the average of three determinations, and the agonistic activity is expressed as

(n=3).

Table 2 The half effective concentration of derivatives 2 and 6 (EC ₅₀ , mM)

为三次测定的平均值，EC₅₀表达为

(n＝3)。Note: The tested concentration range of compound 1-6 is 0.02-1.52 μM;

is the mean of three determinations, and the _EC50 is expressed as

(n=3).

3. Conclusion: The experimental results show that the determined rutabine derivatives have strong agonistic activity on MT ₁ and 5-HT _1A receptors, and have potential regulatory or therapeutic effects related to melatonin and receptors.

Formulation Example 1:

The rutabine derivatives 1-6 prepared according to the method of Preparation Example 1, respectively or mixed with a small amount of DMSO to dissolve, conventionally add water for injection, fine filtration, potting and sterilization to prepare an injection solution.

Formulation Example 2:

According to the method of Preparation Example 1, first prepare the derivatives 1-6 of rutabine, separately or mix them, dissolve them with a small amount of DMSO, dissolve them in sterile water for injection, stir to dissolve, and use a sterile suction filter funnel to dissolve them. Filtration, then sterile fine filtration, sub-packaging in ampoules, freeze-drying at low temperature, aseptic melting and sealing to obtain powder injection.

Formulation Example 3:

According to the method of Preparation Example 1, first prepare the rudbine derivatives 1-6, separate or mix, add excipients in a ratio of 9:1 by weight to excipients, and prepare powders.

Formulation Example 4:

According to the method of Preparation Example 1, firstly prepare and obtain Pterostilbene derivatives 1-6, separately or mixed, add excipients in a ratio of 5:1 by weight to excipients, and granulate and press into tablets.

Formulation Example 5:

According to the method of Preparation Example 1, the rudbine derivatives 1-6 obtained first were prepared separately or mixed, and made into an oral liquid according to the conventional oral liquid preparation method.

Formulation Example 6:

According to the method of Preparation Example 1, the rudbine derivatives 1-6 were prepared separately or mixed, and the excipients were added in a ratio of 5:1 by weight to the excipients to prepare capsules.

Formulation Example 7:

According to the method of Preparation Example 1, the rudbine derivatives 1-6 were prepared separately or mixed, and the excipients were added in a ratio of 3:1 by weight to the excipients to prepare capsules.

Formulation Example 8:

According to the method of Preparation Example 1, the rudbine derivatives 1-6 obtained first were prepared separately or mixed, and the excipients were added in a ratio of 5:1 by weight to the excipients to prepare granules.

Claims (4)

1. the application of the pharmaceutical composition of rubaline derivatives 2 and 6 shown in the following structural formula in the preparation of melatonin and serotonin receptor agonist,

2. the application of the pharmaceutical composition of the rubelline derivatives 2 and 6 shown in the following structural formula in the preparation of the medicine for treating or improving the central nervous system disease relevant to melatonin and serotonin receptors,

3. the application of rubelline derivatives 2 and 6 shown in the following structural formulas in the preparation of a medicine for treating or improving the central nervous system disease relevant to melatonin and serotonin receptors,

4. the application of rubaline derivatives 2 and 6 shown in the following structural formula in the preparation of melatonin and serotonin receptor agonist,

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