CN107382730A - A kind of derivative of lourerin B and its preparation and application - Google Patents
A kind of derivative of lourerin B and its preparation and application Download PDFInfo
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Abstract
The invention discloses a kind of derivative of lourerin B and its preparation and application.The derivative of the lourerin B, there is structure as shown below：.The present invention has carried out modification transformation to lourerin B structure, the derivative of designed production is in addition to the analgesic activities of lourerin B to be retained, its structural stability will be significantly larger than lourerin B, and block the related voltage-gated potassium channel Kv1.3 of autoimmune disease activity also to significantly improve.
Description
Technical field
The invention belongs to medicinal application field, and in particular to a kind of derivative of lourerin B and its preparation and application.
Background technology
Pain great threat human health, has a strong impact on patients ' life quality.Current clinically most commonly used antalgesic
Thing is m orphine compound, and the maximum side effect of this kind of analgesic for acting on opiate receptor is easily to cause user's habituation,
Caused withrawal symptom is greatly to injure to patient after drug withdrawal.Exploitation nothing or weak additive, the clear and definite analgesic of target spot are
The focus in medical research and development field, such analgesic also have larger potential market Development volue.Voltage gated sodium channels
Nav1.7 selective expressions are in mammal peripheral sensory neuron, and after blocking the passage, then periphery algesiroreceptor is pressed down
Make so as to which pain sensation information can not be transmitted to maincenter, and because Nav1.7 does not express in central nervous system, so Nav1.7 spy
Different in nature blocking agent does not cause the additive side effect of patient, Nav1.7 sodium channels still in addition to it can play preferable analgesic activities
By the internationally recognized excellent target for aching medicine without additive town for exploitation.
Gated 1.7 sodium channel protein of mammal Primary Sensory Neuron tetraodotoxin sensitive voltage is internationally recognized
Analgesic effect novel targets.
Lourerin B is derived from a kind of chalcone compounds of dai medicine Resina Draconis, studies have found that lourerin B has very
Strong analgesic activities, due to the presence of phenolic hydroxyl group in molecular structure, lourerin B phenolic hydroxyl group is caused easily to be oxidized to quinoid structure,
Cause lourerin B structural stability poor, have impact on its pharmacological activity.Lourerin B is mainly from dai medicine Resina Draconis at present in addition
Extraction, source are limited, and extraction lourerin B is also unfavorable for the protection to rare plant resource dragon tree.
The content of the invention
In order to solve the above-mentioned technical problem, the present invention has carried out modification transformation to lourerin B structure, designed production
For derivative in addition to the analgesic activities of lourerin B to be retained, its structural stability will be significantly larger than lourerin B.
The invention provides a kind of derivative of lourerin B, it has structure as shown below：
。
The present invention provides purposes of the derivative of above-mentioned lourerin B as Nav1.7 sodium channel blockers.
The derivative that the present invention provides above-mentioned lourerin B is used for the application for preparing town pain medicine.
The present invention provides purposes of the derivative of above-mentioned lourerin B as the immunodepressant of targeting Kv1.3 potassium channels.
The derivative that the present invention provides above-mentioned lourerin B is used to prepare prevention and treatment autoimmune disease medicine
Purposes.
The derivative that the present invention provides above-mentioned lourerin B leads to as Nav1.7 sodium channel blockers and Kv1.3 potassium simultaneously
The purposes of road blocking agent.
The derivative that the present invention provides above-mentioned lourerin B is used for the use for preparing the medicine for the treatment of rheumatoid arthritis
On the way.
The present invention provides the preparation method of the derivative of above-mentioned lourerin B, comprises the following steps：
1）Lourerin B is being reacted in organic solvent with acetylation reagent；Wherein, the acetylation reagent be chloroacetic chloride,
Acetic anhydride, acetic acid and concentrated sulfuric acid mol ratio are 1:0.05 ~ 0.1 mixture or acetic acid-dicyclohexylcarbodiimide mol ratio is
1:1.0 ~ 1.5 mixture；
2）Step 1）The mixture that obtains after completion of the reaction is purified, dry after obtain the derivative of the lourerin B, i.e. acetyl
Change lourerin B.
Preferably, step 1）Described in organic solvent be dichloromethane, chloroform, acetone or carbon tetrachloride in one kind
Or its combination.
Preferably, step 2）For by step 1）Obtained mixture is reacted successively with sodium acid carbonate, saturated aqueous common salt and water
Washing, then with anhydrous sodium sulfate drying, then with column chromatographic isolation and purification, obtains the derivative of the lourerin B.
The present invention combines the structural unstable factor of lourerin B, carries out acetylation to its phenolic hydroxyl group, and provide out one
Brand-new acetylation lourerin B chemical synthesis process, realizes the acetylation modification to lourerin B, and can largely be closed
Into.
The invention provides purposes of the derivative of lourerin B as Nav1.7 sodium channel blockers.The present invention have detected
Influence of the acetylation lourerin B of synthesis to Primary Sensory Neuron voltage gated sodium channels Nav1.7, find acetylation dragon
Sanguinin B blocks Nav1.7 half-inhibition concentration suitable with lourerin B, proves that acetylation lourerin B remains the town of lourerin B
Pain acts on.Acute toxicity test in mice shows that acetylation lourerin B is almost non-toxic, safe to use.
The invention provides purposes of the acetylation lourerin B as the gated potassium channel Kv1.3 of blocking voltage blocking agent.
Acetylation lourerin B is capable of the gated potassium channel Kv1.3 of blocking voltage of concentration dependent, and Kv1.3 channel blockers are to control
Treat the important lead compound of rheumatoid arthritis.Show that acetylation lourerin B has sample simultaneous rheumatoid arthritis
The excellent results controlled, the anti-inflammatory particular by blocking Kv1.3 potassium channels, by blocking Nav1.7 sodium channels to ease pain.
The good characteristic of the more targets of medicine of acetylation lourerin B possessed one provided by the invention, in analgesia, especially controlling
Treat and the medicine in rheumatoid arthritis patient pain's aspect than existing Clinical practice has great advantage.
Brief description of the drawings
Figure 1A is the nucleus magnetic hydrogen spectrum figure of lourerin B produced by the present invention.
Figure 1B is the nuclear-magnetism carbon spectrogram of lourerin B produced by the present invention.
Fig. 2A is the nucleus magnetic hydrogen spectrum figure of acetylation lourerin B produced by the present invention.
Fig. 2 B are the nuclear-magnetism carbon spectrograms of acetylation lourerin B produced by the present invention.
Fig. 3 A show acetylation lourerin B（ES）To the inhibitory action of Nav1.7 sodium channel currents.
Fig. 3 B show acetylation lourerin B（ES）To the amount effect curve of the inhibitory action of Nav1.7 sodium channel currents.
Fig. 3 C show lourerin B（LrB）To the inhibitory action of Nav1.7 sodium channel currents.
Fig. 3 D show lourerin B（LrB）To the amount effect curve of the inhibitory action of Nav1.7 sodium channel currents.
Fig. 4 A show acetylation lourerin B（ES）To the inhibitory action of Kv1.3 channel currents.
Fig. 4 B show acetylation lourerin B（ES）To the amount effect curve of the inhibitory action of Kv1.3 channel currents.
Fig. 4 C show lourerin B（LrB）To the inhibitory action of Kv1.3 channel currents.
Fig. 4 D show lourerin B（LrB）To the amount effect curve of the inhibitory action of Kv1.3 channel currents.
Embodiment
The invention will be further described with specific embodiment below in conjunction with the accompanying drawings, so that those skilled in the art can be with
It is better understood from the present invention and can be practiced, but illustrated embodiment is not as a limitation of the invention.
First, the brand-new synthetic route design of acetylation lourerin B and specific preparation method.
1st, synthetic route is as follows：
Whole synthetic route is reacted including five steps.The first step is with 1,3,5- trimethoxy-benzene（Ⅰ）For raw material, pass through
Vilsmeier-Haauc acylation reactions prepare reaction intermediate 2,4,6- TMBs（Ⅳ）；Second step is with to hydroxyl
Benzoylformaldoxime（Ⅱ）And benzyl chloride（Ⅲ）For raw material, intermediate 4- benzyloxy acetophenones are prepared by nucleophilic substitution（Ⅴ）；
3rd step is with reaction intermediate（Ⅳ）With（Ⅴ）For raw material, by Claisen-Schmidt condensation reactions, reaction intermediate is prepared
2,4,6- -4 '-benzyloxy chalcones of trimethoxy（Ⅵ）；4th step be then by（Ⅵ）Catalytic hydrogenating reduction is 2,4,6- front threes
Epoxide -4 '-hydroxyl dihydrochalcone（Lourerin B, VII）；5th step is then pair（Ⅶ）Acetylation modification is carried out, obtains final production
Thing 2,4,6- trimethoxies -4 '-acetoxyl group dihydrochalcone（Acetylation lourerin B, VIII）.
2nd, specific preparation method：
Step 1：With 1,3,5- trimethoxy-benzene（Ⅰ）For raw material, prepared by Vilsmeier-Haauc acylation reactions in reaction
Mesosome 2,4,6- TMBs（Ⅳ）.
Equipped with agitator and it is being put into the 250ml of ice bath three-necked flask, is adding 8.76g（0.12mol）DMF, then will
9.8g（0.064mol）POCl3It is slowly dropped in three-necked flask, Vilsmeier complexs is formed after reacting 0.5h, then will
10.10g（0.06mol）1,3,5- trimethoxy-benzene is slowly added in this complex reaction liquid.Then ice bath is removed, loads onto condensation
Pipe, after reaction bulb to be moved to 30 DEG C of water-bath the inside stirring reaction 3h generation intermediates, intermediate reaction liquid is poured into and is equipped with
In the beaker of 60ml frozen water, after being stirred and evenly mixed with glass bar, it is put into after beaker mouth seals up sealed membrane in 4 DEG C of refrigerators and cools down analysis
Crystalline substance, next day filter, and obtain beige white powder shape product 8.8g, yield 74.68%.
Step 2：With parahydroxyacet-ophenone（Ⅱ）And benzyl chloride（Ⅲ）For raw material, prepared by nucleophilic substitution middle
Body 4- benzyloxy acetophenones（Ⅴ）.
At room temperature, in the three-necked flask of the 250ml equipped with agitator, DMF is sequentially added（50ml）, parahydroxyacet-ophenone
（10g, 0.0735mol）、K2CO3（20.2g 0.147mol）And benzyl chloride（10.228g 0.0808mol）, then at room temperature
Reaction 16h is mixed, reaction is finished, reactant mixture is poured into the frozen water beaker equipped with 60ml, is cooled down, then filter and sunk
Form sediment, drying can obtain 16.2g white powder target products, yield 98.38%.
Step 3：The reaction intermediate prepared with step 1 and step 2（Ⅳ）With（Ⅴ）For raw material, pass through Claisen-
Schmidt condensation reactions, prepare -4 '-benzyloxy chalcone of trimethoxy of reaction intermediate 2,4,6-（Ⅵ）.
In the 100ml ground three-necked bottles equipped with magnetic stirring apparatus and condenser pipe, 10ml methanol, 4- benzyloxies are sequentially added
Acetophenone（2.26g 0.01mol）, the lower addition of stirring is made into molten by 30ml methanol and potassium hydroxide (2.80g, 0.10mol)
Liquid, 50 DEG C are heated to, after bulk drug dissolving, add 2,4,6- TMBs（1.96g 0.01mol）, heat back
Stream, thin-layer chromatography monitoring reaction are completed until reaction.Stop reaction, it is to be cooled, filter, filter residue and drying, obtain yellow powder and consolidate
Body material 3.60g, as target product, yield 89%.
Step 4：Will（Ⅵ）Catalytic hydrogenating reduction is 2,4,6- -4 '-hydroxyl dihydrochalcones of trimethoxy（Lourerin B,
Ⅶ）.
In the 100ml ground three-neck flasks equipped with magnetic stirring apparatus and condenser pipe, methanol is sequentially added（25ml）With 2,
4,6- -4 '-benzyloxy chalcones of trimethoxy（1.9g, 0.005mol）, lower addition ammonium formate (1.8g, 0.03mol) is stirred,
10%Pd-C（0.2g）And methanol（25ml）, it is heated to flowing back, after the completion of question response, is filtered to remove palladium carbon, filtrate decompression is concentrated to give
To pale yellow oil, proper amount of acetone dissolving is added, is filtered to remove unnecessary ammonium formate, then be concentrated under reduced pressure to give grease, very
Sky is dried, and is obtained yellowish solid material, is used column chromatography to obtain white solid powder i.e. lourerin B 1.11g, yield
70%.Lourerin B, also known as 2,4,6- -4 '-hydroxyl dihydrochalcones of trimethoxy, are white powdery solids.Products therefrom passes through
Nuclear-magnetism, nucleus magnetic hydrogen spectrum such as Figure 1A are detected, nuclear-magnetism carbon is composed as shown in Figure 1B, and spectroscopic data is as follows：
1H-NMR( 400MHz,DMSO-D6):δ2.78( 2H,dd,J=9.5Hz,J=5.8Hz,-CH2Ar), 2.99 (2H,
dd,J=9.6 Hz,J=5.9Hz,-CH2CO ), 3.74( 6H,s,-OCH3 ), 3.76( 3H,s,-OCH3 ), 6.22(
2H,s,H-2,5 ), 6.85( 2H,d,J=8.7Hz,H-3’,5’ ), 7.84( 2H,d,J=8.7Hz,H-2’,6’),
10.32( 1H,s,OH-4’)；
13C-NMR (125MHz, DMSO-d 6 ) δ：198.1（C=O）, 161.9（C-4’）, 159.3（C-2,6）, 158.3（C-4）,
130.5（C-2’,6’）, 128.1（1’）, 115.2（C-3’,5’）, 108.6（1）, 90.7（C-3,5）, 55.6（2,6-OCH3）,
55.2（4-OCH3）, 37.9（C-α）, 18.3（C-β）.
Through nmr analysis, the molecular formula C of step 4 products therefrom18H20O5, its nuclear magnetic spectrum and published lourerin B
Nuclear magnetic spectrum is consistent.
Step 5：It is right（Ⅶ）Acetylation modification is carried out, obtains -4 '-acetoxyl group two of trimethoxy of final product 2,4,6-
Hydrogen chalcone（Acetylation lourerin B, VIII）.
30ml CH are sequentially added in the 250ml ground three-necked flasks equipped with magnetic stirring apparatus2Cl2And lourerin B
(2.53g, 0.001mol), then add 70ml CH2Cl2, a few drop pyridines, which are added dropwise, makes solution become clarification, then with constant pressure liquid separation
CH is added dropwise in funnel3COCl（6.28g 0.08mol）, flow control is in 1 drop/min, thin-layer chromatography monitoring reaction to reaction stopping.
Reaction is finished, and reaction solution is poured into separatory funnel, washed successively with 10% sodium acid carbonate（25ml×2）, saturated common salt water washing
（25ml×2）, distill water washing（25ml×2）, then by organic layer anhydrous sodium sulfate drying, filter, filtrate decompression is concentrated to give
To pale yellow oil crude product, finally crude product is used column chromatography to obtain 1.52g whites stringy solids i.e. target compound, produced
Rate 53%.The final product VIII of synthesis is white fluffy solid, molecular formula C2OH22O6, its nucleus magnetic hydrogen spectrum figure such as Fig. 2A, nuclear-magnetism carbon
As shown in Figure 2 B, its spectroscopic data is as follows for spectrogram：
1H-NMR(600MHz,DMSO-d6):δ2.30(3H,s,-COCH3), 2.80(2H,m,H-β), 3.00(2H,d,J=
8.7Hz,H-α), 3.75(6H,s,-OCH3), 3.76(3H,s,-OCH3), 6.22(2H,s,H-3.5), 7.28(2H,m,H-
3’,5’), 8.01(2H,d,J=8.7Hz,H-2’,6’)
13C-NMR(600MHz,DMSO-d6):δC198.9(C=O), 38.3(C-α), 18.1(C-β), 108.2(C-1),
158.2(C-2),90.2(C-3),154.0(C-4),90.7(C-5),158.2(C-6),134.0(C-1’),129.6(C-2’),
122.2(C-3’),159.4(C-4’),122.2(C-5’),129.6(C-6’),55.6(2,6-OCH3),55.2(4-OCH3),
168.4(C-OAC),20.9(CH3-OAC)
It was found from nuclear magnetic spectrogram analysis, this step products therefrom is as follows for the molecular formula of acetylation lourerin B：
。
Second, the acute toxicity test of upper laxative remedy measure acetylation lourerin B
Operating method and step：The limiting test of 2000mg/kg dosage levels：Tested material is given to a mouse respectively.If
The dead mouse, then carry out master trip；If the mouse survival, two kinds of medicines are given to other 4 mouse, animal respectively successively
Sum is 5.If the phase is dead after the test for an animal, and other animals survive, and should stop that other animals are administered, to institute
There is animal to be observed, if also to occur during similar observation dead.The animal of later death should be dynamic with other death
Thing is equally counted, and result is evaluated as follows：When having three or more than three animal deads, LD50 is less than 2000mg/kg；
When having 3 or more than 3 animal survivals, LD50 is more than 2000mg/kg；If three animal deads, then master trip is carried out.
Result of the test：From five female KM mouse（Disease Control and Prevention Center of Hubei Province provides）, according to upper every animal of laxative remedy
To mouse be injected intraperitoneally the acetylation lourerin B of synthesis according to 2000mg/kg dosage levels, the results showed that tested five
Experiment mice without death condition, thinks that the safety in utilization of acetylation lourerin B is higher according to experimental method.
3rd, using suppression of the patch clamp experiments technology for detection acetylation lourerin B to mammal Nav1.7 sodium channels
Effect
Mammal Nav1.7 sodium channels are excellent target of the research and development without additive town pain medicine, and Nav1.7 specific inhibition agents are
Without additive town pain medicine lead compound, this research intend using patch-clamp electro physiology experiment detection acetylation lourerin B with
Blocking effect of the lourerin B to mammal Nav1.7 sodium channels.
HEK293-T cell lines do not express Nav1.7 sodium channel proteins in itself, be Study of Exogenous Nav1.7 channel designs with
The good host cell of function.This experiment by HEK293T cells (ATCC ACS4500) cultivate added with 10% hyclone,
100 units/ml penicillin, 100 μ g/ml streptomysins DMEM culture mediums in (Life Technologies,
GrandIsland, NY, USA), cell is placed on 37 °C and 5% CO2Cultivated in incubator.Someone Nav1.7 sodium channels will be contained
The carrier pGFP-C1-SCN9A of encoding gene（Buy in OriGene Technologies companies of the U.S., article No. NM-002977）
Transfect and give HEK293T cells, electrophysiology is used in transfection 24-48 hours and is tested.It is used to record Nav1.7 sodium channels in experiment
The extracellular fluid component of electric current（mmol/L）For：5 KCl, 140 NaCl, 10 Hepes, 2 CaCl2, 1 MgCl2, 10
D-glucose, it is 7.4 to adjust outer liquid pH value with NaOH.Liquid composition in electrode（mmol/L）For：140 KCl, 1 MgCl2, 1
EGTA, 3 Na2ATP, 10 Hepes, pH value is adjusted to 7.2 with KOH.Experimental drug is purchased from Sigma companies (St.
Louis, MO, USA).The lourerin B and acetylation lourerin B of various concentrations are dissolved in for recording Nav1.7 sodium channels electricity
In the extracellular fluid of stream.Whole-cell recording is carried out using EPC10 amplifier systems, environment temperature maintains 22-24 °C,
The application of the setting of experiment parameter, the collection of data and stimulation is controlled by Patch master softwares, and wave filter 1 is set
For 10kHz（Bessel）, wave filter 2 is arranged to 2.9kHz（Bessel）.Quartz glass wool embryonic tube（BF 150-86-10;
Sutter companies, the U.S.) through P-97 drawing instrument（Sutter companies, the U.S.) level drawing, electrode resistance is after charging interior liquid
2-4 MΩ.To record with after glass microelectrode slightly malleation, by means of MP225 narishiges（Sutter companies, the U.S.）Move
Move glass microelectrode and progressively close to cell, after release malleation between electrode and cell membrane in favor of forming high resistance seals, enter
The fast electric capacity of row compensates automatically（c-fast）, slightly after negative pressure rupture of membranes, then capable slow electric capacity compensates automatically（c-slow）And series resistance
Compensation（R-series）.Cell membrane potential is clamped down in -60mV under whole cell voltage clamp recording pattern, given within every 10 seconds
100ms step-lengths ,+10mV depolarising square wave stimulate Nav1.7 sodium channel currents on active cell film, observe in extracellular fluid not
The Nav1.7 sodium channel currents of recombination expression are influenceed with concentration acetylation lourerin B and lourerin B（With the cell of not drug containing
Outer liquid is as control（control））.As a result as shown in Fig. 3 A ~ 3D, acetylation lourerin B is thin in HEK293-T to heterogenous expression
Nav1.7 channel currents on after birth show the inhibitory action similar with lourerin B, and concentration dependent feature is presented.Pass through
Hill equations are fitted to the amount effect curve of acetylation lourerin B, obtain its Nav1.7 sodium channel to external source recombination expression
Electric current half-inhibition concentration is 4.33 ± 0.15 μM, suitable with 3.97 ± 0.08 μM of half-inhibition concentrations of lourerin B.Show dragon
The derivative acetylation lourerin B that sanguinin B obtains through acetylation modification is effectively maintained while structural stability is increased
The town pain activity of lourerin B, what acetylation lourerin B can develop into targeting Nav1.7 sodium channels aches medicine without additive town.
4th, made using suppression of the patch clamp experiments technology for detection acetylation lourerin B to mammal Kv1.3 potassium channels
With
Mammal Kv1.3 types potassium channel blocker can suppress the immune response that body increases extremely, be exploitation treatment itself
The lead compound of immunity disease medicine.Because lourerin B can block mammal Kv1.3 type potassium channels, this research is intended
By lourerin B derivative acetylation lourerin B of the diaphragm device experiment detection through acetylation modification and lourerin B to mammal
The blocking effect of Kv1.3 type potassium channels.
HEK293-T cell lines not expressing K v1.3 channel proteins in itself, are Study of Exogenous Kv1.3 channel designs and function
Good host cell.This experiment cultivates HEK293T cells (ATCC ACS4500) added with 10% hyclone, 100
Unit/ml penicillin, 100 μ g/ml streptomysins DMEM culture mediums in (Life Technologies, GrandIsland,
NY, USA), cell is placed on 37 °C and 5% CO2Cultivated in incubator.By carrier pSP64（Taught by Stephan Grissmer
Award present, University of Ulm, Ulm, Germany）The cDNA of middle coding mKv1.3 channel proteins is through XhoI/BamH
I multiple cloning sites are subcloned into pIRES2- EGFP (Clontech, Inc., Mountain View, CA, USA) carrier
In.The clone of structure is analyzed by DNA sequencing to determine the correctness of its nucleotide sequence.With Lipofectamine 2000
(Invitrogen) carrier of structure is transfected and gives HEK293T cells, electrophysiology is used for after 24 hours and is tested.Used in experiment
In the extracellular fluid component of record Kv1.3 channel currents（mmol/L）For：5 KCl, 140 NaCl, 10 Hepes, 2
CaCl2, 1 MgCl2, 10 D-glucose, it is 7.4 to adjust outer liquid pH value with NaOH.Liquid composition in electrode（mmol/L）For：
140 KCl, 1 MgCl2, 1 EGTA, 3 Na2ATP, 10 Hepes, pH value is adjusted to 7.2 with KOH.Experimental drug is purchased from
Sigma companies (St. Louis, MO, USA).Whole-cell recording, environment temperature are carried out using EPC10 amplifier systems
Degree maintains 22-24 °C, the application of the setting of experiment parameter, the collection of data and stimulation by Patch master softwares come
Control, wave filter 1 are arranged to 10kHz（Bessel）, wave filter 2 is arranged to 2.9kHz（Bessel）.Quartz glass wool embryonic tube（BF
150-86-10;Sutter companies, the U.S.) through P-97 drawing instrument（Sutter companies, the U.S.) level drawing, charge interior liquid
Electrode resistance is 2-4 M Ω afterwards.After forming high resistance seals between electrode and cell membrane, carry out fast electric capacity and compensate automatically（c-
fast）, slightly after negative pressure rupture of membranes, then capable slow electric capacity compensates automatically（c-slow）And series resistance compensation（R-series）.Complete
Cell membrane potential is clamped down in -60mV under cell voltage pincers logging mode, gives within every 10 seconds 400ms step-lengths ,+50mV goes to pole
Changing square wave stimulates Kv1.3 channel currents on active cell film, observes various concentrations acetylation lourerin B and dragon's blood in extracellular fluid
Plain B influences on the Kv1.3 channel currents of recombination expression（With the extracellular fluid for being used to record Kv1.3 channel currents of not drug containing
As comparison liquid（control）And eluent（wash）.Elution：HEK293T cells after aforementioned bearer transfection are containing acetylation
After acetylation lourerin B being observed in the extracellular fluid of lourerin B the Kv1.3 channel currents of recombination expression being influenceed, with without
The extracellular fluid of medicine elutes to the extracellular fluid of the lourerin B containing acetylation, and observation acetylation lourerin B blocks Kv1.3
Whether the effect of channel current is reversible.）.As a result as shown in Fig. 4 A ~ 4D, comparison diagram 4A ~ Fig. 4 D, it can be seen that acetylation lourerin B
The inhibitory action more stronger than lourerin B is shown to Kv1.3 channel current of the heterogenous expression on HEK293-T cell membranes, and is in
Existing concentration dependent feature, inhibitory action can be eluted, and be shown to be invertibity effect.By Hill equations to acetylation dragon's blood
Plain B amount effect curve is fitted, obtain its Kv1.3 channel currents half-inhibition concentration to external source recombination expression for 2.10 ±
0.39 μ Μ, it is better than 7.19 ± 0.59 μM of half-inhibition concentrations of lourerin B.Show that lourerin B spreads out through what acetylation modification obtained
Biological acetylation lourerin B is also improved while structural stability is increased to the depression effect of Kv1.3 passages.Acetyl
Change lourerin B can develop into targeting Kv1.3 potassium channels immunodepressant, for autoimmune disease prevention and control
Treat.
In summary the experimental result of research approach three and four, show that the acetylation lourerin B that the present invention synthesizes will controlled
Treating in rheumatoid arthritis has big advantage, is in particular in acetylation lourerin B by blocking Kv1.3 potassium channels,
For suppressing the inflammatory reaction of rheumatoid arthritis, play a part of " effecting a permanent cure "；By blocking Nav1.7 sodium channels, improve and suffer from
The pain sensation symptom of person, plays a part of " taking stopgap measures ".
Embodiment described above is only to absolutely prove preferred embodiment that is of the invention and being lifted, protection model of the invention
Enclose not limited to this.The equivalent substitute or conversion that those skilled in the art are made on the basis of the present invention, in the present invention
Protection domain within.Protection scope of the present invention is defined by claims.
Claims (10)
1. a kind of derivative of lourerin B, it is characterised in that it has structure as shown below：
Explanation:Explanation: E:Gwssi CPC clients cases inventions 42f8325d-9946-417b-bb1a-05b1c5d9def8 new 100001 641400dest_path_image002.jpg。
2. purposes of the derivative of the lourerin B described in claim 1 as Nav1.7 sodium channel blockers.
3. the derivative of the lourerin B described in claim 1 is used for the application for preparing town pain medicine.
4. purposes of the derivative of the lourerin B described in claim 1 as the immunodepressant of targeting Kv1.3 potassium channels.
5. the derivative of the lourerin B described in claim 1 is used for the use for preparing prevention and treatment autoimmune disease medicine
On the way.
6. the derivative of the lourerin B described in claim 1 hinders as Nav1.7 sodium channel blockers and Kv1.3 potassium channels simultaneously
The purposes of disconnected agent.
7. the derivative of the lourerin B described in claim 1 is used for the purposes for preparing the medicine for the treatment of rheumatoid arthritis.
8. the preparation method of the derivative of the lourerin B described in claim 1, it is characterised in that comprise the following steps：
1）Lourerin B is being reacted in organic solvent with acetylation reagent；Wherein, the acetylation reagent be chloroacetic chloride,
Acetic anhydride, acetic acid and concentrated sulfuric acid mol ratio are 1:0.05 ~ 0.1 mixture or acetic acid-dicyclohexylcarbodiimide mol ratio is
1:1.0 ~ 1.5 mixture；
2）Step 1）The mixture that obtains after completion of the reaction is purified, dry after obtain the derivative of the lourerin B, i.e. acetyl
Change lourerin B.
9. preparation method according to claim 8, it is characterised in that step 1）Described in organic solvent be dichloromethane,
One kind or its combination in chloroform, acetone or carbon tetrachloride.
10. preparation method according to claim 8, it is characterised in that step 2）For by step 1）React obtained mixing
Thing is successively with sodium acid carbonate, saturated aqueous common salt and water washing, then with anhydrous sodium sulfate drying, then with column chromatographic isolation and purification,
Obtain the derivative of the lourerin B.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115124409A (en) * | 2022-05-30 | 2022-09-30 | 山东科源制药股份有限公司 | Preparation method of propafenone hydrochloride intermediate |
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| CN101200420A (en) * | 2007-11-07 | 2008-06-18 | 上海大学 | Method for preparing 4'-hydroxy-2,4,6-trimethoxy dihydrocharcone |
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| CN115124409A (en) * | 2022-05-30 | 2022-09-30 | 山东科源制药股份有限公司 | Preparation method of propafenone hydrochloride intermediate |
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