CN107382730B - 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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- CN107382730B CN107382730B CN201710519342.8A CN201710519342A CN107382730B CN 107382730 B CN107382730 B CN 107382730B CN 201710519342 A CN201710519342 A CN 201710519342A CN 107382730 B CN107382730 B CN 107382730B
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Abstract
The invention discloses a kind of derivative of lourerin B and its preparations and application.The derivative of the lourerin B has structure as follows：.The present invention has carried out modification transformation to lourerin B structure, the derivative of designed production is other than the analgesic activities of lourerin B to be retained, its structural stability will be significantly larger than lourerin B, and the activity of the relevant voltage-gated potassium channel Kv1.3 of autoimmune disease is blocked also to significantly improve.
Description
Technical field
The invention belongs to medicinal application fields, and in particular to a kind of derivative of lourerin B and its preparation and application.
Background technology
Pain great threat human health, seriously affects patients ' life quality.Current clinically most commonly used antalgesic
Object is m orphine compound, and this kind of maximum side effect of the analgesic for acting on opiate receptor is easily to cause user's habituation,
The withrawal symptom generated after drug withdrawal is greatly to injure to patient.Exploitation nothing or weak additive, the specific analgesic of target spot are
The hot spot 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 channel, then periphery algesiroreceptor is pressed down
It makes that pain sensation information cannot be transmitted to maincenter, and since Nav1.7 is not expressed in central nervous system, so the spy of Nav1.7
Anisotropic blocking agent does not cause the additive side effect of patient, the sodium channels Nav1.7 still other than it can play preferable analgesic activities
By the internationally recognized excellent target for aching drug without additive town for exploitation.
Gated 1.7 sodium channel protein of mammal Primary Sensory Neuron tetraodotoxin sensitive voltage is internationally recognized
Analgesic act on 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 cause lourerin B phenolic hydroxyl group to be easy to be oxidized to quinoid structure due to the presence of phenolic hydroxyl group in molecular structure,
Cause lourerin B structural stability poor, affects its pharmacological activity.In addition current lourerin B is mainly from dai medicine Resina Draconis
Extraction, source is limited, and extraction lourerin B is also unfavorable for the protection to rare plant resource dragon tree.
Invention content
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 other than the analgesic activities of lourerin B to be retained, structural stability will be significantly larger than lourerin B.
The present invention provides a kind of derivatives of lourerin B, with structure as follows：
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 to prepare the application that drug is ached in town.
The present invention provides purposes of the derivative of above-mentioned lourerin B as the immunosuppressor for 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 drug
Purposes.
The present invention provides the derivative of above-mentioned lourerin B while leading to as Nav1.7 sodium channel blockers and Kv1.3 potassium
The purposes of road blocking agent.
The present invention provide above-mentioned lourerin B derivative be used to prepare treatment rheumatoid arthritis drug use
On the way.
The present invention provides the preparation method of the derivative of above-mentioned lourerin B, includes the following steps：
1）Lourerin B is being reacted in organic solvent with acetylation reagent；Wherein, the acetylation reagent is acetyl
Chlorine, acetic anhydride, acetic acid and concentrated sulfuric acid molar ratio are 1:0.05 ~ 0.1 mixture or acetic acid-dicyclohexylcarbodiimide molar ratio
It is 1:1.0 ~ 1.5 mixture；
2）Step 1）The mixture that obtains after completion of the reaction is purified, it is dry after obtain the derivative of the lourerin B, i.e.,
Acetylation lourerin B.
Preferably, step 1）Described in organic solvent be dichloromethane, chloroform, acetone or carbon tetrachloride in one kind
Or combinations thereof.
Preferably, step 2）For by step 1）Mixture obtained by the reaction is successively with sodium bicarbonate, saturated salt solution and water
Washing, is then dried with anhydrous sodium sulfate, then with column chromatographic isolation and purification, obtain the derivative of the lourerin B.
The structural unstable factor of present invention combination lourerin B carries out acetylation to its phenolic hydroxyl group, and provides out one
Completely new acetylation lourerin B chemical synthesis process, realizes the acetylation modification to lourerin B, and can largely be closed
At.
Purposes the present invention provides the derivative of lourerin B as Nav1.7 sodium channel blockers.The present invention has detected
Influence of the acetylation lourerin B of synthesis to Primary Sensory Neuron voltage gated sodium channels Nav1.7 finds acetylation dragon
Sanguinin B blocks the half-inhibition concentration of Nav1.7 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.
Purposes the present invention provides acetylation lourerin B as the blocking agent of the gated potassium channel Kv1.3 of blocking voltage.
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, it is anti-inflammatory particular by blocking Kv1.3 potassium channels, eased pain by blocking the sodium channels Nav1.7.
The good characteristic for the more targets of a medicine that acetylation lourerin B provided by the invention has in analgesia, is especially being controlled
It treats and the drug in rheumatoid arthritis patient pain's aspect than existing Clinical practice has great advantage.
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 inhibiting effect of Nav1.7 sodium channel currents.
Fig. 3 B show acetylation lourerin B（ES）To the amount effect curve of the inhibiting effect of Nav1.7 sodium channel currents.
Fig. 3 C show lourerin B（LrB）To the inhibiting effect of Nav1.7 sodium channel currents.
Fig. 3 D show lourerin B（LrB）To the amount effect curve of the inhibiting effect of Nav1.7 sodium channel currents.
Fig. 4 A show acetylation lourerin B（ES）To the inhibiting effect of Kv1.3 channel currents.
Fig. 4 B show acetylation lourerin B（ES）To the amount effect curve of the inhibiting effect of Kv1.3 channel currents.
Fig. 4 C show lourerin B（LrB）To the inhibiting effect of Kv1.3 channel currents.
Fig. 4 D show lourerin B（LrB）To the amount effect curve of the inhibiting effect of Kv1.3 channel currents.
Specific implementation mode
The invention will be further described in the following with reference to the drawings and specific embodiments, 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.
One, the completely new synthetic route design of acetylation lourerin B and specific preparation method.
1, synthetic route is as follows：
Entire 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- trimethoxybenzaldehyde（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）；
Third is walked with reaction intermediate（Ⅳ）With（Ⅴ）Reaction intermediate is prepared by Claisen-Schmidt condensation reactions for raw material
2,4,6- -4 '-benzyloxy chalcones of trimethoxy（Ⅵ）；4th step be then by（Ⅵ）Catalytic hydrogenating reduction is 2,4,6- front threes
Oxygroup -4 '-hydroxyl dihydrochalcone（Lourerin B, VII）；5th step is then pair（Ⅶ）Acetylation modification is carried out, final production is obtained
Object 2,4,6- trimethoxies -4 '-acetoxyl group dihydrochalcone（Acetylation lourerin B, VIII）.
2, specific preparation method：
Step 1：With 1,3,5- trimethoxy-benzene（Ⅰ）For raw material, prepared by Vilsmeier-Haauc acylation reactions anti-
Answer intermediate 2,4,6- trimethoxybenzaldehyde（Ⅳ）.
It equipped with blender and is being put into the three-necked flask of 250ml of ice bath, 8.76g is added（0.12mol）DMF, then will
9.8g（0.064mol）POCl3It is slowly dropped in three-necked flask, forms Vilsmeier complexs 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, condensation is loaded onto
Reaction bulb is moved to inside 30 DEG C of water-bath after being stirred to react 3h generation intermediates, intermediate reaction liquid is poured into and is equipped with by pipe
In the beaker of the ice water of 60ml, after being stirred and evenly mixed with glass bar, cooling analysis is put into 4 DEG C of refrigerators after beaker mouth seals up sealed membrane
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 intermediate
Body 4- benzyloxy acetophenones（Ⅴ）.
At room temperature, in the three-necked flask of the 250ml equipped with blender, 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 finishes, reaction mixture is poured into the ice water beaker equipped with 60ml, cooling, then filters and sunk
It forms 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 2,4,6- -4 '-benzyloxy chalcone of trimethoxy of reaction intermediate（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）, be added with stirring be made by 30ml methanol and potassium hydroxide (2.80g, 0.10mol) it is molten
Liquid is heated to 50 DEG C, after bulk pharmaceutical chemicals dissolving, adds 2,4,6- trimethoxybenzaldehyde（1.96g 0.01mol）, heat back
Stream, thin-layer chromatography monitoring reaction are completed until reaction.Stop reaction, it is to be cooled, it filters, filter residue and drying, it is solid to obtain yellow powder
Body substance 3.60g, as target product, yield 89%.
Step 4：It 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）, it is added with stirring ammonium formate (1.8g, 0.03mol),
10%Pd-C（0.2g）And methanol（25ml）, it is heated to flowing back, waits after the completion of reacting, be 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 extra ammonium formate, then be concentrated under reduced pressure to give grease, very
Sky is dry, obtains yellowish solid material, uses 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 is detected, nucleus magnetic hydrogen spectrum such as Figure 1A, nuclear-magnetism carbon spectrum is 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, nuclear magnetic spectrum and published lourerin B
Nuclear magnetic spectrum is consistent.
Step 5：It is right（Ⅶ）Acetylation modification is carried out, 2,4,6- -4 '-acetoxyl group two of trimethoxy of final product is obtained
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 adds 70ml CH2Cl2, a few drop pyridines, which are added dropwise, makes solution become clarification, then uses 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, is washed successively with 10% sodium bicarbonate（25ml×2）, saturated common salt water washing
（25ml×2）, distill water washing（25ml×2）, then organic layer dried with anhydrous sodium sulfate, it filters, 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, be produced
Rate 53%.The final product VIII of synthesis is white fluffy solid, molecular formula C2OH22O6, nucleus magnetic hydrogen spectrum figure such as Fig. 2A, nuclear-magnetism carbon
Spectrogram is as shown in Figure 2 B, and spectroscopic data is as follows：
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)
From nuclear magnetic spectrogram analysis it is found that this step products therefrom is as follows for the molecular formula of acetylation lourerin B：
Second, upper laxative remedy measures the acute toxicity test of 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, carries out master trip；If the mouse survival, two kinds of drugs are given to other 4 mouse respectively successively,
Animal number 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,
All animals are observed, if also occur during similar observation dead.The animal of later death should be with other death
Animal equally count, result is evaluated as follows：When having three or three or more animal deads, LD50 is less than 2000mg/
kg；When having 3 or 3 or more animal survivals, LD50 is more than 2000mg/kg；If there is three animal deads, then main examination is carried out
It tests.
Test result：Select five female KM mouse（Disease Control and Prevention Center of Hubei Province provides）, according to every animal of upper laxative remedy
The acetylation lourerin B of intraperitoneal injection synthesis is carried out to mouse according to 2000mg/kg dosage levels, the results showed that tested five
Experiment mice thinks that the safety in utilization of acetylation lourerin B is higher without death condition according to experimental method.
Third, the inhibition using patch clamp experiments technology detection acetylation lourerin B to the sodium channels mammal Nav1.7
Effect
The sodium channels mammal Nav1.7 are to research and develop the excellent target that drug is ached without additive town, Nav1.7 specific inhibitions
Agent is that the lead compound of drug is ached without additive town, this research is quasi- using patch-clamp electro physiology experiment detection acetylation dragon's blood element
The blocking effect of B and lourerin B to the sodium channels mammal Nav1.7.
HEK293-T cell strains itself do not express Nav1.7 sodium channel proteins, 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% fetal calf serum,
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% CO2It is cultivated in incubator.The sodium channels someone Nav1.7 will be contained
The carrier pGFP-C1-SCN9A of encoding gene（It buys in OriGene Technologies companies of the U.S., article No. NM-002977）
It transfects and gives HEK293T cells, transfect in 24-48 hours and tested for electrophysiology.For recording the sodium channels Nav1.7 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 external solution pH value with NaOH.Liquid ingredient in electrode（mmol/L）For：140 KCl, 1 MgCl2, 1
EGTA, 3 Na2ATP, 10 Hepes adjust pH value 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 concentration are dissolved in for recording the sodium channels Nav1.7 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 acquisition of data and stimulation is controlled by Patch master softwares, and filter 1 is arranged
For 10kHz（Bessel）, filter 2 is set as 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 positive pressure, by means of MP225 narishiges（Sutter companies, the U.S.）It moves
Dynamic glass microelectrode and gradually close to cell, after release positive pressure is in favor of the formation high resistance seals between electrode and cell membrane, into
The fast capacitance of row compensates automatically（c-fast）, slightly after negative pressure rupture of membranes, then capable slow capacitance 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, is given within every 10 seconds
100ms step-lengths ,+10mV depolarising square wave stimulation active cell film on Nav1.7 sodium channel currents, observe extracellular fluid in not
The Nav1.7 sodium channel currents of recombinant expression are influenced with concentration acetylation lourerin B and lourerin B（With the cell of not drug containing
External solution is as a contrast（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 inhibiting effect similar with lourerin B, and concentration dependent feature is presented.Pass through
Hill equations are fitted the amount effect curve of acetylation lourerin B, obtain its sodium channel Nav1.7 recombinantly expressed to external source
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 is obtained through acetylation modification is effectively maintained while increasing structural stability
The town pain activity of lourerin B, what acetylation lourerin B can develop into targeting Nav1.7 sodium channels aches drug without additive town.
4th, acetylation lourerin B is detected using patch clamp experiments technology, the inhibition of mammal Kv1.3 potassium channels is made
With
Mammal Kv1.3 types potassium channel blocker can inhibit the immune response that body increases extremely, be exploitation treatment
The lead compound of autoimmune disease drug.Since lourerin B can block mammal Kv1.3 type potassium channels, originally grind
The quasi- lourerin B derivative acetylation lourerin B for passing through the experiment detection of diaphragm device through acetylation modification and lourerin B are studied carefully to lactation
The blocking effect of animal Kv1.3 type potassium channels.
HEK293-T cell strains not expressing K v1.3 channel proteins itself, are Study of Exogenous Kv1.3 channel designs and function
Good host cell.This experiment cultivates HEK293T cells (ATCC ACS4500) added with 10% fetal calf serum, 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% CO2It is cultivated in incubator.By carrier pSP64（It is 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, tested for electrophysiology after 24 hours.It is 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 external solution pH value with NaOH.Liquid ingredient in electrode（mmol/L）For：
140 KCl, 1 MgCl2, 1 EGTA, 3 Na2ATP, 10 Hepes adjust pH value 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 acquisition of data and stimulation by Patch master softwares come
Control, filter 1 are set as 10kHz（Bessel）, filter 2 is set as 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, carries out fast capacitance and compensate automatically（c-
fast）, slightly after negative pressure rupture of membranes, then capable slow capacitance 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
Change Kv1.3 channel currents on square wave stimulation active cell film, observes various concentration acetylation lourerin B and dragon's blood in extracellular fluid
Plain B influences the Kv1.3 channel currents of recombinant expression（With the extracellular fluid for recording Kv1.3 channel currents of not drug containing
Liquid as a contrast（control）And eluent（wash）.Elution：HEK293T cells after aforementioned bearer transfection are containing acetylation
After observing that acetylation lourerin B influences the Kv1.3 channel currents of recombinant expression in the extracellular fluid of lourerin B, to be free of
The extracellular fluid of drug elutes 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
Inhibiting effect 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, inhibiting effect can be eluted, and be shown to be invertibity effect.By Hill equations to acetylation dragon's blood
The amount effect curve of plain B is fitted, obtain the Kv1.3 channel current half-inhibition concentrations that it recombinantly expresses external source be 2.10 ±
0.39 μ Μ are 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 the depression effect in the channels Kv1.3 while increasing structural stability.Acetyl
Change lourerin B can develop into targeting Kv1.3 potassium channels immunosuppressor, for autoimmune disease prevention and control
It treats.
In summary the experimental result of research approach three and four shows that the acetylation lourerin B that the present invention synthesizes will controlled
Treat rheumatoid arthritis on have big advantage, be in particular in acetylation lourerin B by block Kv1.3 potassium channels,
Inflammatory reaction for inhibiting rheumatoid arthritis is played the role of " effecting a permanent cure "；By blocking the sodium channels Nav1.7, improves and suffer from
The pain sensation symptom of person, plays the role 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
It encloses without being limited thereto.Those skilled in the art on the basis of the present invention made by equivalent substitute or transformation, in the present invention
Protection domain within.Protection scope of the present invention is subject to claims.
Claims (4)
1. purposes of the derivative of lourerin B as the immunosuppressor of targeting Kv1.3 potassium channels；
Wherein, the derivative of the lourerin B has structure as follows：
2. the derivative of lourerin B is used to prepare the purposes of prevention and treatment autoimmune disease drug；
Wherein, the derivative of the lourerin B has structure as follows：
3. the derivative of lourerin B while the purposes as Nav1.7 sodium channel blockers and Kv1.3 potassium channel blockers；Its
In, the derivative of the lourerin B has structure as follows：
4. the derivative of lourerin B is used to prepare the purposes of the drug for the treatment of rheumatoid arthritis；
Wherein, the derivative of the lourerin B has structure as follows：
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