CN109456335A - Aoxidize the synthetic method of bicuculline - Google Patents
Aoxidize the synthetic method of bicuculline Download PDFInfo
- Publication number
- CN109456335A CN109456335A CN201811453273.6A CN201811453273A CN109456335A CN 109456335 A CN109456335 A CN 109456335A CN 201811453273 A CN201811453273 A CN 201811453273A CN 109456335 A CN109456335 A CN 109456335A
- Authority
- CN
- China
- Prior art keywords
- compound
- heated
- bicuculline
- synthetic method
- added
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/06—Peri-condensed systems
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of synthetic methods for aoxidizing bicuculline, specifically with 3,4- dimethoxyphenylacetic acid is starting material, it reacts products therefrom with bromine to react with homopiperony lamine again, products therefrom is restored with reducing agent again later with phosphorus oxychloride cyclization, products therefrom and BOC anhydride reaction, products therefrom adds Lithium Aluminium Hydride after reacting with thricyclohexyl phosphorus and palladium acetate and is reacted, and finally reacts again with manganese acetate (III) to get oxidation bicuculline crude product is arrived.The present invention provides a kind of new methods by fully synthetic preparation oxidation bicuculline, and the synthetic method route is simple, yield height (45% or more).
Description
Technical field
The present invention relates to the synthetic methods of oxidation bicuculline, belong to pharmaceutical technology field.
Background technique
Bicuculline is mainly derived from bloodroot leaf lotus bud tree peony waterborne (Dicentra spectabilis), prevents
Section plant bleedingheart stephania herb (Stephania dicentrinifera), alias Dicentrine or Dicentrine.There is researcher
By isolated method, it is raw that biologically active aporphine, isoquinolin and bisisoquinoline are isolated from O.leucoxylon
Alkaloids, the bicuculline that these compounds include, the bioactivity with antitumor, antalgic and sedative effect and antibacterial.S- lotus
Luxuriant bicuculline can be isolated from Lindera megaphylla (Lindera megaphylla (Lauraceae)), passes through what is carried out to it
It is antitumor the study found that it has more significant anti-tumor activity to human hepatocytes tumor HuH-7.S- bicuculline is to coming from
In the external activity research of the human tumor cell in the 20 of seven kinds of different tissues a kind of, all tumour cells are shown
Different degrees of anti-tumor activity, IC50It is worth range by 0.4 μM to esophageal cancer cell HCE-6 to 29 to hepatoma HA22T
μM, it is demonstrated experimentally that S- bicuculline to various cancer cells have certain inhibitory activity (Huang R L, Chen C C,
Huang Y L,Ou J C,Hu CP,Chen C F,Chang C.Anti-Tumor Effects of d-Dicentrine
from the Root of Lindera megaphyll.Planta Med,1998,64(3),212-215.).Such as adrenal gland
The activity of plain receptor acting, ion channel action activity, cytotoxicity, antioxidant activity, platelet aggregation inhibitory activity and anti-cone
Worm activity etc..
The bicuculline (dicentrinone, i.e. oxidation bicuculline, abbreviation DCO) of oxidation, has and kills Li Shiman
The activity of (original) worm and trypanosome, DCO is to 52 repair-deficient of Trypanocidal, Antileishmanial and rad
RS322 yeast strain has preferable inhibitory activity.Meanwhile the inhibitor that bicuculline is Topoisomerase I, II is aoxidized,
And its (Zhou B, Johnson R K, Mattern M R, Wang X, Hecht S also weaker to the toxicity of cell
M.Isolation and biochemical characterization of a new topoisomerase I
inhibitor from Ocotea leucoxylon.J.Nat.Prod.2000,63,217–221.Stevigny C,Bailly
C,Quetin-Leclercq.Cytotoxic and Antitumor Potentialities of Aporphinoid
Alkaloids.Curr.Med.Chem.-Anti-Cancer Agents, 2005,5 (2), 173-182 (10)), get a good chance of
Anti-cancer drugs as great potential.But since DCO alkaloid is extremely low in nature content, it is difficult to meet structure-activity relationship and activity
Pharmacological research etc. needs, and therefore, in order to sufficiently develop latent effect of the DCO in terms of medicine, is obtained by the method for organic synthesis
It is highly important to DCO.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of synthetic methods of the high oxidation bicuculline of yield.
The synthetic method of oxidation bicuculline of the present invention is former for starting with 3,4- dimethoxyphenylacetic acid
Material, is synthesized by following synthetic route:
Wherein, the reducing agent is selected from one or more of sodium borohydride, lithium borohydride and potassium borohydride
Combination.
Above-mentioned oxidation bicuculline more specifically synthetic method the following steps are included:
1) synthesis of compound (2):
It takes 3,4- dimethoxyphenylacetic acid to be dissolved in glacial acetic acid, bromine is added and is reacted, gained reactant pours into ice water
In, it stands, filter, obtain compound (2);
2) synthesis of compound (3):
It takes compound (2) to be placed in thionyl chloride, is reacted under the conditions of being heated or not heated, reactant is evaporated off unreacted
Thionyl chloride obtains weak yellow liquid;Homopiperony lamine is taken to be dissolved in the first organic solvent, acquired solution is added to aforementioned faint yellow
It in liquid, is reacted under the conditions of being heated or not heated, solvent is evaporated off in reactant, obtains compound (3);
3) synthesis of compound (4):
It takes compound (3) to be placed in the first organic solvent, phosphorus oxychloride is added, is reacted under the conditions of being heated or not heated,
Solvent is evaporated off in reactant, obtains compound (4);
4) synthesis of compound (5):
It takes compound (4) to be dissolved in the first organic solvent, excessive reducing agent is added, it is anti-under the conditions of being heated or not heated
It answers, diluted acid is added into gained reactant to remove unreacted reducing agent, resulting material is extracted with extractant, and collection has
Machine phase, is washed with saturated sodium bicarbonate solution, dry, obtains compound (5);
5) synthesis of compound (6):
Compound (5) is taken to be dissolved in the first organic solvent, two dimethyl dicarbonate fourths are added to alkalinity in the pH value of regulation system
Ester reacts under the conditions of being heated or not heated, and gained reactant is extracted with extractant, collects organic phase, is spin-dried for, is changed
It closes object (6);
6) synthesis of compound (7):
Thricyclohexyl phosphorus and palladium acetate is taken to be dissolved in the second organic solvent, the pH=8-10 of regulation system is added thereto
Compound (6), reacts under atmosphere protection and heating condition, and gained reactant is extracted with extractant again after being neutralized with acid,
Organic phase is collected, is washed with saturated sodium bicarbonate solution, it is dry, obtain compound (7);
7) synthesis of compound (8):
It takes compound (7) to be dissolved in tetrahydrofuran, Lithium Aluminium Hydride is added under condition of ice bath, it is anti-under the conditions of atmosphere protection
It answers, adjusts the pH=8-9 of gained reactant, filter, collect filtrate, filtrate is spin-dried for, and obtains compound (8);
8) synthesis of bicuculline, that is, compound (9) is aoxidized:
It takes compound (8) to be dissolved in glacial acetic acid, manganese acetate (III) (C is added6H9MnO6.2(H2O)), in being heated or not heated
Under the conditions of react, gained reactant filtering, collect filtrate, solvent is evaporated off, obtain oxidation bicuculline crude product;
In above-mentioned synthetic method, first organic solvent be selected from chloroform, methylene chloride, methanol, ethyl alcohol, propyl alcohol and
The combination of one or more of n-butanol;Second organic solvent be n,N-Dimethylformamide (DMA) and/or
DMAC N,N' dimethyl acetamide (DMAC);The extractant is selected from one of chloroform, methylene chloride and ethyl acetate.
In above-mentioned synthetic method:
In step 1), the molar ratio of 3, the 4- dimethoxyphenylacetic acid and bromine is stoichiometric ratio, in practical operation
In usually to choose the molar ratio of 3,4- dimethoxyphenylacetic acid and bromine be 1:1.2-1:1.5;Reaction preferably carries out at normal temperature,
Whether reaction can be used thin-layer chromatography tracing detection completely, and under above-mentioned qualifications, reaction to taking around 1-3h completely.It should
In step, the dosage of glacial acetic acid can determine as needed, it is generally the case that with 3, the 4- dimethoxyphenylacetic acid of 0.1mol for base
Standard, whole raw materials are preferably dissolved with the glacial acetic acid of 180-250mL.It is preferred that 3 are then added to after bromine is first dissolved with glacial acetic acid,
It is reacted in the glacial acetic acid solution of 4- dimethoxyphenylacetic acid.The yield of this step is 95% or more.
In step 2), it is also reactant that thionyl chloride, which had both made solvent, and the molar ratio of compound (2) and thionyl chloride is usually
1:1.5-2.0;The compound (2) and thionyl chloride are preferably performed under heating conditions, and are more preferably returned 60 to solvent
Back flow reaction is carried out under the conditions of stream temperature, whether reaction can be used thin-layer chromatography tracing detection completely, under above-mentioned qualifications,
Reaction to taking around 3-5h completely.The yield of this step is 80% or more.
In step 3), the molar ratio of compound (3) and phosphorus oxychloride is stoichiometric ratio, is usually chosen in actual operation
The molar ratio of compound (3) and phosphorus oxychloride is 1:4-1:5;The compound (3) and phosphorus oxychloride are preferably in heating condition
Lower progress, more preferably carries out back flow reaction under 60 DEG C to solvent of reflux conditions, and whether reaction can be used thin layer completely
Tracing detection is chromatographed, under above-mentioned qualifications, reaction to taking around 2-4h completely.The yield of this step is 80% or more.
In step 4), the reaction preferably carries out under normal temperature conditions, and reaction to taking around 8-12h completely.It is described
Diluted acid be usually 0.5-2mol/L hydrochloric acid solution.The yield of this step is 85% or more.
In step 5), the molar ratio of compound (5) and di-tert-butyl dicarbonate (BOC acid anhydrides) is stoichiometric ratio, in reality
The molar ratio that compound (5) and di-tert-butyl dicarbonate are usually chosen in the operation of border is 1:1.0-1:1.5;Reaction is preferably in room temperature
Under the conditions of carry out, reaction to taking around 3-5h completely.The yield of this step is 95% or more.
In step 6), the molar ratio of compound (6) and thricyclohexyl phosphorus is stoichiometric ratio, in actual operation usually choosing
Taking the molar ratio of compound (6) and thricyclohexyl phosphorus is 1:1-1:1.2;Reaction more preferably carries out under the conditions of 120-135 DEG C,
Under above-mentioned qualifications, reaction to taking around 20-30h completely.In the step, palladium acetate makees catalyst use, dosage
Preferably 2% or more of compound (6) quality, the more preferably 2.5-3% of compound (6) quality.The yield of this step exists
82% or more.
Step 5), 6) and 7) in, the aqueous solution for generalling use existing common alkaline matter carrys out the pH value of regulation system, excellent
Choosing is adjusted using wet chemical, sodium hydrate aqueous solution, ammonium hydroxide or triethylamine etc..In step 5), body is preferably adjusted
PH >=8 of system, the more preferably pH=8.5-10 of regulation system.
In step 7), the molar ratio of compound (7) and Lithium Aluminium Hydride is stoichiometric ratio, is usually chosen in actual operation
The molar ratio of compound (7) and Lithium Aluminium Hydride is 1:8-1:10;Reaction is more preferably performed under heating conditions, and is more preferably existed
Back flow reaction under the conditions of 60-70 DEG C, under above-mentioned qualifications, reaction to taking around 20-30h completely.The yield of this step
90% or more.
In step 8), manganese acetate (III) makees catalyst use, and dosage is preferably the 2 times or more of compound (8) quality, more
Preferably 3-4 times of compound (8) quality.The reaction of the compound (8) and manganese acetate (III) is preferably in a heated condition
It carries out, more preferably back flow reaction under the conditions of 60-70 DEG C, under above-mentioned qualifications, reaction to taking around 4-6h completely.
The yield of this step is 50% or more.
In above-mentioned steps 1) -7) in, resulting compound is crude product, in order to reduce the impurity for being introduced into subsequent step,
The purity for improving object crude product simultaneously, is preferably used further to subsequent operation for gained compound after purification.Described is pure
Change specifically can be gained crude compound is recrystallized with solvent after be used further to subsequent operation, the solvent recrystallized
It can be the mixture that methanol and/or ethyl alcohol or water and methanol or ethyl alcohol are formed by the volume ratio of 1:2-3.
Similarly, in order to improve the purity for aoxidizing bicuculline, preferably after obtaining oxidation bicuculline crude product again
Carry out purification step, specific purification step are as follows: by silica gel column chromatography on oxidation bicuculline crude product, with by chloroform and first
Alcohol, or eluted by methylene chloride and methanol by the mixed solvent that the volume ratio of 30-35:1 forms, eluent solvent evaporated,
Up to oxidation bicuculline.By limiting the ratio of eluant, eluent, quickly and accurately object is eluted.
Compared with prior art, the present invention provides a kind of new sides by fully synthetic preparation oxidation bicuculline
Method, this method synthetic route is simple to operation, and yield is high (in terms of object after purification, yield is 45% or more).
Detailed description of the invention
Fig. 1 is the crystal structure figure (removing H atom) of final product made from the embodiment of the present invention 1;
Fig. 2 is the gel electrophoresis figure of oxidation bicuculline and DNA effect of the present invention.
Specific embodiment
The present invention is described in further detail combined with specific embodiments below, content to better understand the invention, but
The present invention is not limited to following embodiments.
Embodiment 1
1) synthesis of compound (2):
By 72g 3,4- dimethoxyphenylacetic acid is dissolved in the glacial acetic acid of 600mL, and 7.2g is added after stirring 1h under room temperature
Glacial acetic acid (60mL) solution of bromine after the reaction was continued 2h, is added 200mL ice water, there is white precipitate generation, filter, filter cake is used
After recrystallizing methanol, 96g compound (2) are obtained, yield is about 95%.
Compound (2) is white solid, ESI-MS m/z 273.02 [(2)-H]-,13C-NMR (500MHz, DMSO) δ:
41.0816 (C-2), 56.2538 (C-5), 56.4330 (C-6), 114.9042 (C-9), 115.7982 (C-3), 115.9616
(C-8), 127.4302 (C-10), 148.5825 (C-4), 148.9687 (C-8), 172.0429 (C-1),1H-NMR(500MHz,
DMSO) δ: 3.6021 (2H, S, H-2), 3.7120 (3H, S, H-5), 3.7346 (3H, S, H-6), 6.9880 (1H, S, H-3),
7.0886(1H,S,H-8)。
2) synthesis of compound (3):
100g compound (2) is dissolved in the thionyl chloride of 100mL, flow back 1.5h under the conditions of 76 DEG C, and vacuum distillation is fallen not
There is reacted thionyl chloride, obtain flaxen liquid, dissolves 75g homopiperony lamine with 400mL methylene chloride, and slowly by it
It is added in above-mentioned weak yellow liquid, stirring at normal temperature 4 hours, removes methylene chloride under reduced pressure, with recrystallizing methanol, it is white about to obtain 100g
Color solid, yield are about 80%.
Compound (3) is white solid, ESI-MS m/z 421.96 [(3)+H]+;1H-NMR(500MHz,CDCl3) δ:
2.485 (2H, S, H-8), 3.269 (2H, S, H-12), 3.408 (2H, S, H-9), 2.485 (2H, S, H-8), 3.697 (6H, S,
H-19,20), 6.332 (1H, S, H-15), 6.374 (1H, S, H-4), 6.490 (1H, S, H-7), 6.611 (1H, S, H-5),
6.831(H,S,H-7)。13C-NMR(500MHz,CDCl3) δ: 34.828 (C-12), 40.487 (C-8), 43.350 (C-9),
55.884 (C-19, C-20), 100.5927 (C-1), 107.965 (C-4), 108.704 (C-5), 113.524 (C-14),
114.470 (C-15), 115.434 (C-16), 121.286 (C-7), 126.336 (C-13), 132.033 (C-6), 145.849
(C-3), 147.468 (C-17), 148.525 (C-2), 148.745 (C-18), 169.440 (C-11).
3) synthesis of compound (4):
The compound (3) of 100g is dissolved in 1250mL chloroform, 180mL phosphorus oxychloride (POCl is then added3), reflux is anti-
3h is answered, vacuum distillation removes solvent after the reaction was completed, is washed with saturated sodium bicarbonate solution, and it is dry, obtain unpurified compound
(4), yield about 80%.
4) synthesis of compound (5):
70g compound (4) is dissolved in 300mL methanol, is added excessive sodium borohydride (80g), stirring at normal temperature reaction
12h is slowly added to the dilute hydrochloric acid solution of 1mol/L to react excessive sodium borohydride, and resulting material is extracted with ethyl acetate,
Organic phase is washed with saturated sodium bicarbonate solution, and again with methanol recrystallization obtains about 78g compound (5), yield about 82%.
5) synthesis of compound (6):
The compound (5) of 50g is dissolved in 375mL chloroform, the sodium hydroxide water that 150mL concentration is 2mol/L is then added
Solution stirs the BOC acid anhydrides of the amount of substances such as being slowly added to after 0.5h (pH=9 of system at this time), continues to stir 4h, uses chlorine
Imitative extraction, organic phase are washed with saturated sodium bicarbonate solution again, are evaporated under reduced pressure later, obtain 57g compound (6), yield is about
96%.
6) synthesis of compound (7):
The palladium acetate of the thricyclohexyl phosphorus of 0.32g, 3.15g potassium carbonate and 0.125g are dissolved in (this in the dry DMF of 80mL
When system pH=9), then be added 5g compound (6), inert gas (helium) protection under, in 135 DEG C reflux for 24 hours, then
It is cooling, it after being neutralized with the hydrochloric acid of 1mol/L, is extracted with chloroform, saturated sodium bicarbonate solution washing is dry, after ethyl alcohol recrystallization
4.3g compound (7) are obtained, yield is about 92%.
7) synthesis of compound (8):
The compound (7) of 5g is dissolved in the dry tetrahydrofuran of 130mL, the aluminum hydride of 4.4g is slowly added in ice bath
Lithium flows back for 24 hours in 50 DEG C under inert gas (helium) protection, with the pH=8 of weak aqua ammonia regulation system, filters while hot, collects
Filtrate, vacuum distillation remove solvent, and gained residue ethyl alcohol recrystallization filters after solid is precipitated, obtains compound (8) about
3.5g, yield are about 90%.
8) synthesis of bicuculline, that is, compound (9) is aoxidized:
It takes 2g compound (8) to be dissolved in the glacial acetic acid of 50mL, 6.5g manganese acetate (III) then is added, next time in 70 DEG C of conditions
5h is flowed, reactant filters, and filtrate revolving removes solvent, and residue is to aoxidize bicuculline crude product.Gained crude product chloroform
Dissolution, is washed three times with saturated sodium bicarbonate solution later, then upper silica gel column chromatography (washing away remaining catalyst), with by chlorine
The mixed solvent elution that imitative and methanol is formed by the volume ratio of 30-32:1, eluent solvent evaporated to get high-purity yellow powder
Last product 1g, yield about 50%.
The present embodiment products therefrom is taken to be dissolved in the in the mixed solvent being made of chloroform and methanol by the volume ratio of 3:1, room temperature
Lower slowly volatilization, at the 15th day, discovery had bright yellow rhabdolith to occur, and selects suitable monocrystalline and carries out structural characterization:
1) IR Characterization:
With the Spectrum Two FT-IR Spectrometer Fourier Transform Infrared Spectroscopy of Perkin-Klmer company
Instrument (KBr tabletting) carries out infrared analysis to product made from the present embodiment, and gained ir data is as follows:
IR(KBr cm-1) (Ar-H) 3377 (m), (- CH2-, v) 2923,2851 (m), (C=O) 1638 (vs), (C=C)
1598,1572,1515,1424(s),(-CH2-,d)1459,(C-O)1279,1248,(C-N)1057,(-CH2-)776cm-1。
2) X-ray diffraction is analyzed:
It takes sizeable monocrystalline to be placed in the face Smart Aapex2CCD Bruker to visit on single crystal diffractometer, with graphite monochromatic
The Mo-K alpha ray of device monochromatizationIn the case where temperature is 296 (2) K, using Crystalclear program,
In the range of 1.90 °≤θ≤25.10 °, crystal data is collected with ψ/θ scanning mode, structure cell is determined after least square refinement
Parameter solves crystal structure by direct method and difference Fourier synthetic method, and is modified with complete matrix least square method,
The anisotropic temperature factor of whole non-hydrogen atoms is corrected, and hydrogen is added by theoretical calculation.All calculating are in PC machine
It is upper to be completed using SHELXTL-97 program bag, using semi-empirical method correcting structure.
The structure chart of gained crystal is as shown in Figure 1, its crystal parameters and part bond distance, bond angle data are such as following respectively
Shown in Tables 1 and 2.
Table 1:
Table 2:
Accordingly, it can be determined that the present embodiment products therefrom is that target product aoxidizes bicuculline.
Embodiment 2
Embodiment 1 is repeated, unlike:
Step 3) -5) in, the first organic solvent being related to is changed to ethyl alcohol;
In step 4), reducing agent is changed to lithium borohydride;
In step 5), the pH=8 of regulation system, extractant uses chloroform instead;
In step 6), the pH=10 of regulation system, the dosage of palladium acetate is changed to the 3% of compound (6) quality, and second is organic
Solvent is changed to DMAC, and extractant uses methylene chloride instead;
In step 7), the pH=9 of regulation system;
In step 8), the dosage of manganese acetate (III) is changed to 4 times of compound (8) quality, elution when upper silicagel column elution
Agent is to be eluted by methylene chloride and methanol by the mixed solvent that the volume ratio of 32-35:1 forms.
The present embodiment products therefrom is dissolved in the in the mixed solvent being made of chloroform and methanol by the volume ratio of 3:1, room temperature
Lower slowly volatilization, carries out IR Characterization for the crystal of precipitation and single crystal diffraction is analyzed, and is determined as target product oxidation lotus bud tree peony
Alkali.
Embodiment 3
Embodiment 1 is repeated, unlike:
In step 3), the first organic solvent being related to is changed to n-butanol;
In step 4), the first organic solvent being related to is changed to normal propyl alcohol, and reducing agent is changed to potassium borohydride;
In step 5), the first organic solvent being related to is changed to methanol, the pH=10 of regulation system;
In step 6), the pH=10 of regulation system, the dosage of palladium acetate is changed to the 2% of compound (6) quality, and second is organic
Solvent is changed to DMAC, and extractant uses ethyl acetate instead;
In step 7), the pH=8.5 of regulation system;
In step 8), the dosage of manganese acetate (III) is changed to 2 times of compound (8) quality, elution when upper silicagel column elution
Agent is to be eluted by methylene chloride and methanol by the mixed solvent that the volume ratio of 30:1 forms;
Step 1) -7) in, solvent when being related to recrystallization operation is changed to second alcohol and water by the combination of the volume ratio of 2:1
Object.
The present embodiment products therefrom is dissolved in the in the mixed solvent being made of chloroform and methanol by the volume ratio of 3:1, room temperature
Lower slowly volatilization, carries out IR Characterization for the crystal of precipitation and single crystal diffraction is analyzed, and is determined as target product oxidation lotus bud tree peony
Alkali.
Experimental example 1: oxidation bicuculline tests the proliferation inhibition activity of a variety of human tumor cell lines
The cell strain that this experiment is selected are as follows: bladder cancer cell line T-24, human liver cancer cell HepG2, oophoroma SKOV-3, ovum
Nest cancer persister cell SKOV-3-DDP.Influence with mtt assay evaluation drug to living cell growth and proliferation.
In Primary Screening Test, the panel of tumor cell strain in logarithmic growth phase is taken, with the training for containing 10% newborn bovine serum
Nutrient solution is made into individual cells suspension, with every 190 μ L (about 1 × 10 of hole4A/hole) in 96 orifice plates, culture 12h is waited for carefully cell inoculation
After born of the same parents are adherent, every hole is separately added into the 10 μ L of sample of various concentration, and each gradient sets 4 multiple holes in parallel, and DMSO therein is to help
Solvent, ultimate density are no more than 1%, while setting corresponding negative control group and (there was only cell and equivalent DMSO, no medicine in culture solution
Object) and blank control group (there was only the drug of equivalent in culture solution, cell-free), each gradient also sets 4 multiple holes in parallel, and drug is made
It is 48 hours with the time.Culture terminates first 4 hours every holes and 10 μ L MTT (5mg/mL PBS) is added, and after continuing culture 4 hours, inhales
Supernatant is abandoned, 100 hole μ L/ DMSO is added, 10min is vibrated with plate shaker, allows crystal sufficiently to dissolve, blank pair
According to a group zeroing.Absorbance (A) value after removal background absorbance value is measured with 570nm/630nm dual wavelength with microplate reader, is pressed
It states formula and calculates cell proliferation inhibition rate:
Inhibiting rate=(1- sample A value/control group A value) × 100%
All experiments are averaged after being repeated 3 times.As a result as described in Table 3.
Table 3 (unit %):
Experimental example 2: effect of the oxidation bicuculline to pUC19 Plasmid DNA
With tbe buffer liquid (0.08mol/L Tris-HCl, pH=8.5,0.08mol/L boric acid, 0.0024mol/L
EDTA the Ago-Gel solution for) preparing 1%, is then added a certain amount of GelRed nucleic acid dye;
PUC19 Plasmid DNA is acted on and is tested: adding 0.5 μ L pUC19 Plasmid DNA (0.5 μ g/ μ L) to cross to sterilization treatment micro-
In plastic centrifuge tube, then add the compound of various concentration respectively, is settled to 25 μ L with Tris buffer, it is anti-at 37 DEG C of constant temperature
After answering 3h, 3 μ L bromophenol blues are added and are uniformly mixed, loading, cue mark when using bromophenol blue solution as electrophoresis.In 80V/cm voltage
Under, electrophoresis 80 minutes, finally irradiation development in the UV lamp.
Fig. 2 be compound DCO and DNA effect gel electrophoresis figure, as shown in Figure 2, with compound DCO concentration by
Cumulative big, when concentration is 100 μM, the supercoil configuration of DNA is reduced, and DCO and DNA effect is stronger.With compound DCO concentration
Increase, the Form I of DNA band of swimming relatively lags behind, and shows that compound DCO is embedded between DNA spiral base-pair, and occur therewith
Insertion action increases the relative molecular weight of DNA, and swimming band mobility is caused to reduce.Compared with pure dna swims band (swimming lane 1), chemical combination
The mobility of objects system (swimming lane 2-5) decreases, and hysteresis occurs.This is compound and the auxiliary of Insertion action occurs for DNA
Corroborative evidence is bright.
Claims (9)
1. aoxidize bicuculline synthetic method, it is characterised in that: with 3,4- dimethoxyphenylacetic acid be starting material, by with
Lower synthetic route is synthesized:
Wherein, the reducing agent is the group selected from one or more of sodium borohydride, lithium borohydride and potassium borohydride
It closes.
2. synthetic method according to claim 1, it is characterised in that: specific synthetic method the following steps are included:
1) synthesis of compound (2):
It takes 3,4- dimethoxyphenylacetic acid to be dissolved in glacial acetic acid, bromine is added and is reacted, gained reactant is poured into ice water, quiet
It sets, filter, obtain compound (2);
2) synthesis of compound (3):
It takes compound (2) to be placed in thionyl chloride, is reacted under the conditions of being heated or not heated, unreacted dichloro is evaporated off in reactant
Sulfoxide obtains weak yellow liquid;Homopiperony lamine is taken to be dissolved in the first organic solvent, acquired solution is added to aforementioned weak yellow liquid
In, it is reacted under the conditions of being heated or not heated, solvent is evaporated off in reactant, obtains compound (3);
3) synthesis of compound (4):
It takes compound (3) to be placed in the first organic solvent, phosphorus oxychloride is added, is reacted under the conditions of being heated or not heated, react
Solvent is evaporated off in object, obtains compound (4);
4) synthesis of compound (5):
It takes compound (4) to be dissolved in the first organic solvent, excessive reducing agent is added, is reacted under the conditions of being heated or not heated,
Diluted acid is added into gained reactant to remove unreacted reducing agent, resulting material is extracted with extractant, is collected organic
Phase is washed with saturated sodium bicarbonate solution, dry, obtains compound (5);
5) synthesis of compound (6):
Compound (5) is taken to be dissolved in the first organic solvent, di-tert-butyl dicarbonate is added in the pH value of regulation system to alkalinity, in
It is reacted under the conditions of being heated or not heated, gained reactant is extracted with extractant, is collected organic phase, is spin-dried for, obtains compound
(6);
6) synthesis of compound (7):
Thricyclohexyl phosphorus and palladium acetate is taken to be dissolved in the second organic solvent, chemical combination is added in the pH=8-10 of regulation system thereto
Object (6), reacts under atmosphere protection and heating condition, and gained reactant is extracted with extractant again after being neutralized with acid, collects
Organic phase is washed with saturated sodium bicarbonate solution, dry, obtains compound (7);
7) synthesis of compound (8):
It takes compound (7) to be dissolved in tetrahydrofuran, Lithium Aluminium Hydride is added under condition of ice bath, reacted under the conditions of atmosphere protection, adjusted
The pH=8-9 of gained reactant is saved, is filtered, filtrate is collected, filtrate is spin-dried for, and obtains compound (8);
8) synthesis of bicuculline, that is, compound (9) is aoxidized:
It takes compound (8) to be dissolved in glacial acetic acid, is added manganese acetate (III), is reacted under the conditions of being heated or not heated, gained reaction
Object filtering, collects filtrate, solvent is evaporated off, and obtains oxidation bicuculline crude product;
In above-mentioned synthetic method, first organic solvent is selected from chloroform, methylene chloride, methanol, ethyl alcohol, propyl alcohol and positive fourth
The combination of one or more of alcohol;Second organic solvent is n,N-Dimethylformamide (DMA) and/or N, N-
Dimethyl acetamide (DMAC);The extractant is selected from one of chloroform, methylene chloride and ethyl acetate.
3. synthetic method according to claim 2, it is characterised in that: further include the purifying step for aoxidizing bicuculline crude product
Suddenly, specific purification step are as follows: by silica gel column chromatography on oxidation bicuculline crude product, pressed with by chloroform or methylene chloride and methanol
The mixed solvent elution of the volume ratio composition of 30-35:1, eluent solvent evaporated is to get oxidation bicuculline.
4. synthetic method according to claim 2, it is characterised in that: in step 1) -7) in, resulting compound carries out pure
Subsequent operation is used further to after change.
5. synthetic method according to claim 4, it is characterised in that: the purifying be by gained compound solvent into
Subsequent operation is used further to after row recrystallization.
6. synthetic method according to claim 5, it is characterised in that: the solvent be methanol and/or ethyl alcohol, either
The mixture that water and methanol or ethyl alcohol are formed by the volume ratio of 1:2-3.
7. the synthetic method according to any one of claim 2-6, it is characterised in that: in step 6), react in 120-135
It is carried out under the conditions of DEG C.
8. the synthetic method according to any one of claim 2-6, it is characterised in that: in step 6), the dosage of palladium acetate
It is 2% or more of compound (6) quality.
9. the synthetic method according to any one of claim 2-6, it is characterised in that: in step 8), manganese acetate (III)
Dosage is the 2 times or more of compound (8) quality.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811453273.6A CN109456335B (en) | 2018-11-30 | 2018-11-30 | Method for synthesizing oxidized bicuculline |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811453273.6A CN109456335B (en) | 2018-11-30 | 2018-11-30 | Method for synthesizing oxidized bicuculline |
Publications (2)
Publication Number | Publication Date |
---|---|
CN109456335A true CN109456335A (en) | 2019-03-12 |
CN109456335B CN109456335B (en) | 2021-07-23 |
Family
ID=65612135
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811453273.6A Active CN109456335B (en) | 2018-11-30 | 2018-11-30 | Method for synthesizing oxidized bicuculline |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN109456335B (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104387399A (en) * | 2014-12-12 | 2015-03-04 | 广西师范大学 | Synthetic method of oxidized nantenine and application of oxidized nantenine |
-
2018
- 2018-11-30 CN CN201811453273.6A patent/CN109456335B/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104387399A (en) * | 2014-12-12 | 2015-03-04 | 广西师范大学 | Synthetic method of oxidized nantenine and application of oxidized nantenine |
Non-Patent Citations (3)
Title |
---|
BO ZHENG,等: "Novel total syntheses of oxoaporphine alkaloids enabled by mild Cu-catalyzed tandem oxidation/aromatization of 1-Bn-DHIQs", 《RSC ADVANCES》 * |
OM V.SINGH,等: "Manganese(III) acetate mediated oxidation of aporphines: a convenient and useful synthesis of oxoaporphines", 《TETRAHEDRON LETTERS》 * |
RAVI JANGIR,等: "Total Synthesis of Tetrahydroisoquinoline-Based Bioactive Natural Products Laudanosine, Romneine, Glaucine, Dicentrine, and Their Unnatural Analogues Isolaudanosine and Isoromneine", 《SYNTHESIS》 * |
Also Published As
Publication number | Publication date |
---|---|
CN109456335B (en) | 2021-07-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110878046A (en) | Rare earth complex constructed based on 2-methyl-5, 7-dichloro-8-hydroxyquinoline and preparation method and application thereof | |
CN106632424A (en) | Copper chloride complex using 1-(2-pyridine)-9-hexyl-beta-carboline as ligand and synthesis method and application thereof | |
CN109456335A (en) | Aoxidize the synthetic method of bicuculline | |
CN109503609A (en) | Bromo aoxidizes bicuculline and its synthetic method and application | |
CN108125962B (en) | Application of benzo [ d ] aza-quinazoline compound in preparation of drugs for treating lung cancer | |
CN109651413B (en) | Rare earth complex with bromo-oxidized bicuculline as ligand and synthetic method and application thereof | |
CN108329300B (en) | Nitrobenzo [ d ] aza-quinazoline compound and preparation method and application thereof | |
CN106478692A (en) | Copper-nitrate complex and its synthetic method and application with 1 (2 pyridine) 9 benzyl β carboline as part | |
CN106632420A (en) | Copper chloride complex by taking 1-(2-pyridine)-9-butyl-beta-carboline as ligand as well as synthetic method and application of complex | |
CN106632421A (en) | Copper nitrate complex of 1-(2-pyridine)-9-(4-methylbenzyl)-beta-carboline and synthesizing method and application of copper nitrate complex | |
CN109320534B (en) | Oxidized bicuculline rare earth complex and synthesis method and application thereof | |
CN108129460B (en) | Methoxyphenylbenzo [ d ] aza-quinazoline compound and preparation and application thereof | |
CN103044326A (en) | 5-bromo oxoisoaporphine, and synthesis method and application thereof | |
CN108623638B (en) | 12-chlorobenzimidazole-1, 8-naphthalimide-platinum complex and preparation method and application thereof | |
CN109021021B (en) | 11, 12-dichlorobenzimidazole-1, 8-naphthalimide-platinum complex and preparation method and application thereof | |
CN108727434B (en) | 11-trifluoromethyl benzimidazole-1, 8-naphthalimide-platinum complex and preparation method and application thereof | |
CN108078993B (en) | Application of 6-nitroquinazoline compound in preparation of medicine for treating lung cancer | |
CN110437264B (en) | Homocamptothecin 5, 6-dibromo norcantharidinate derivative and regioselective synthesis method thereof | |
CN106478681B (en) | The copper-nitrate complex and its synthetic method of 1- (2- pyridines) -9- (4- luorobenzyls)-B-carboline and application | |
CN106432288B (en) | The copper-nitrate complex of 1 (2 pyridine) 9 (2 Benzyloxyethyl) β carbolines and synthetic method and application | |
CN106632423B (en) | The copper-nitrate complex and its synthetic method of 1- (2- pyridines) -9- (2- phenylethyls)-B-carboline and application | |
CN106478676B (en) | The copper-nitrate complex of 1 (2 pyridine) 9 (2 Phenoxyethyl) β carbolines and synthetic method and application | |
CN106432287B (en) | The copper-nitrate complex of 1 (2 pyridine) 9 (2 ethoxyethyl group) β carbolines and synthetic method and application | |
CN106478686B (en) | Using 1 (2 pyridine) 9 ethyl β carbolines as the copper-nitrate complex and its synthetic method of part and application | |
CN106478678B (en) | The copper-nitrate complex of 1 (2 pyridine) 9 (methyl of naphthalene 2) β carbolines and its synthetic method and application |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |